Patent Publication Number: US-2021163924-A1

Title: Evolution of bont peptidases

Description:
RELATED APPLICATIONS 
     This application claims the benefit under 35 U.S.C. 119(e) of the filing date of U.S. Provisional Application Ser. No. 62/550,408, filed Aug. 25, 2017, entitled “EVOLUTION OF BONT PEPTIDASES”, the entire contents of which are incorporated herein by reference. 
    
    
     FEDERALLY SPONSORED RESEARCH 
     This invention was made with government support under grant numbers EB022376, GM118062 and GM122261 awarded by the National Institutes of Health. The government has certain rights in the invention. 
    
    
     BACKGROUND 
     Over the last few decades, the medical community has witnessed a remarkable shift in the composition of pharmaceutical therapies from traditional small molecules to biomacromolecules. The growing number of macromolecular therapeutics is a result of their potential for highly specific interactions in biological systems and has been facilitated by improvements in molecular biology and biomolecule engineering. Despite their tremendous success, macromolecular therapies have been limited almost exclusively to extracellular targets due to the significant challenge of their controllable delivery into the cytoplasm. While a number of notable advances have been made in the area of macromolecular delivery, this critical problem remains a major barrier to the development and use of macromolecular therapeutics that address intracellular targets. As an alternative, several natural protein systems are capable of cytoplasmic self-delivery. However, the ability to reengineer these systems to imbue them with the necessary binding or catalytic activities and specificities for therapeutic effect is largely underexplored. 
     SUMMARY 
     The disclosure relates to novel Botulinum neurotoxin (BoNT) protease variants and methods of evolving the same. As described herein, BoNT proteases are attractive candidates for continuous evolution because BoNTs provide a built-in cytosolic delivery mechanism, which allows BoNTs to cleave intracellular targets. In some embodiments, evolved BoNT protease variants that cleave a desired substrate (e.g., a disease-associated intracellular protein) are described herein. The disclosure is based, in part, on the discovery that BoNT protease variants that cleave target proteins lacking canonical BoNT cleavage substrates can be produced by phage-assisted continuous evolution (PACE), for example as described in U.S. Pat. No. 9,023,594, issued May 5, 2015, the entire contents of which are incorporated herein by reference. 
     In some embodiments, evolved BoNT protease variants as described herein may be expressed as a part of a full-length toxin comprising of a BoNT light chain (LC) and a BoNT heavy chain (HC). Typically, the catalytic protease domain is located in the light chain (LC) of the BoNT. The BoNT HC encodes domains which generally enable the BoNT protease variant to cross cellular membranes and cleave target proteins in the intracellular environment, making them useful for treating diseases associated with aberrant activity of intracellular proteins (e.g., cancer, neurological disorders, etc.), such as Soluble NSF Attachment Protein Receptors (SNARE) proteins (e.g., VAMP7, VAMPS, etc.). It should be appreciated that evolved BoNT protease variants described herein may comprise an evolved BoNT LC, or both an evolved BoNT LC and HC. In some embodiments, an evolved BoNT protease variant comprises a wild-type BoNT HC. In some embodiments, an evolved BoNT protease variant comprises a BoNT HC having one or more amino acid mutations relative to a wild-type BoNT HC. In some embodiments, the receptor-binding domain of the BoNT HC has been replaced by a protein domain capable of binding to a cell surface receptor or ligand. In some embodiments, this protein domain may take the form of an antibody, lectin, monobody, single-chain variable fragment (scFv), hormone or signaling factor. 
     Accordingly, in some aspects, the disclosure provides a protein that is evolved from a wild-type Botulinum neurotoxin serotype E (e.g., an evolved BoNT E protease variant, also referred to as a “BoNT E variant”) to cleave a non-canonical BoNT E substrate, for example SNAP23 or PTEN, or evolved to cleave the canonical BoNT E substrate SNAP25 with higher efficiency than wild-type BoNT E. In some embodiments, wild-type BoNT E comprises the sequence set forth in SEQ ID NO: 286 
     
       
         
           
               
            
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGT 
               
               
                   
               
               
                 TPQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSG 
               
               
                   
               
               
                 GILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVII 
               
               
                   
               
               
                 MGAEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSIN 
               
               
                   
               
               
                 EFIQDPALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIE 
               
               
                   
               
               
                 EFLTFGGNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYK 
               
               
                   
               
               
                 DIFQEKYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRET 
               
               
                   
               
               
                 YIGQYKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPIT 
               
               
                   
               
               
                 GRGLVKKIIRF*. 
               
            
           
         
       
     
     In some aspects, the disclosure provides a protein comprising an amino acid sequence that is at least 96% identical to SEQ ID NO: 286 (wild-type BoNT E), wherein the protein comprises at least one of the amino acid mutations set forth in Table 1. 
     In some aspects, the disclosure provides a protein that is evolved from a wild-type Botulinum neurotoxin serotype F (e.g., an evolved BoNT F protease variant, also referred to as a “BoNT F variant”) to cleave a non-canonical BoNT F substrate, for example VAMP7, or evolved to cleave a canonical BoNT F substrate (e.g., VAMP1) with higher efficiency than wild-type BoNT F. In some embodiments, wild-type BoNT F comprises the sequence set forth in SEQ ID NO: 287 
     
       
         
           
               
            
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPERN 
               
               
                   
               
               
                 TIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINS 
               
               
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSII 
               
               
                   
               
               
                 LNLLVLGAGPDIFENSSYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYE 
               
               
                   
               
               
                 YTFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKV 
               
               
                   
               
               
                 KQAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATR 
               
               
                   
               
               
                 LSRVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
               
               
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
               
               
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF*. 
               
            
           
         
       
     
     In some aspects, the disclosure provides a protein comprising an amino acid sequence that is at least 90% (e.g., at least 95%, at least 96%, at least 97%, etc.) identical to SEQ ID NO: 287 (wild-type BoNT F), wherein the protein comprises at least one of the amino acid mutations set forth in Table 2. 
     In some aspects, the disclosure relates to evolved BoNT protease variants that cleave intracellular vesicle-associated membrane proteins (VAMPs). VAMPs are members of the SNARE protein family and typically mediate vesicle fusion, such as synaptic vesicle fusion and vesicular secretion. Without wishing to be bound by any particular theory, aberrant function of VAMPs is associated with certain diseases, for example motor neuron disease. Thus, evolved BoNT proteases that cleave certain VAMPs, in some embodiments, are useful for reducing VAMP protein activity inside a cell or a subject. In some embodiments, a protein (e.g., a BoNT F variant) cleaves a vesicle-associated membrane (VAMP) protein, for example, a VAMP7 protein or a VAMP8 protein. In some embodiments, the VAMP7 protein comprises a sequence set forth in SEQ ID NO: 288 
     
       
         
           
               
            
               
                 MAILFAVVARGTTILAKHAWCGGNFLEDFERSRAFNFLNEIKKRFQTTY 
               
               
                   
               
               
                 GSRAQTALPYAMNSEFSSVLAAQLKHHSENKGLDKVMETQAQVDELKGI 
               
               
                   
               
               
                 MVRNIDLVAQRGERLELLIDKTENLVDSSVTFKTTSRNLARAMCMKNLK 
               
               
                   
               
               
                 LTIIIIIVSIVFIYIIVSPLCGGFTWPSCVKK 
               
               
                 or the VAMP8 protein comprises a sequence set 
               
               
                 forth in SEQ ID NO: 289 
               
               
                   
               
               
                 MEEASEGGGNDRVRNLQSEVEGVKNIMTQNVERILARGENLEHLRNKTE 
               
               
                   
               
               
                 DLEATSEHFKTTSQKVARKFWWKNVKMIVLICVIVFIIILFIVLFATGA 
               
               
                   
               
               
                 FS. 
               
            
           
         
       
     
     In some embodiments, a protein (e.g., a BoNT F variant) cleaves a VAMP1 or a VAMP2 protein. In some embodiments, the VAMP1 protein comprises a sequence set forth in SEQ ID NO: 290 
                    MSAPAQPPAEGTEGTAPGGGPPGPPPNMTSNRRLQQTQAQVEEVVDIIR               VNVDKVLERDQKLSELDDRADALQAGASQFESSAAKLKRKYWWKNCKMM               IMLGAICAIIVVVIVRRG,            
or the VAMP2 protein comprises a sequence set forth in SEQ ID NO: 291
 
     
       
         
           
               
            
               
                 MSATAATAPPAAPAGEGGPPAPPPNLTSNRRLQQTQAQVDEVVDIMRVN 
               
               
                   
               
               
                 VDKVLERDQKLSELDDRADALQAGASQFETSAAKLKRKYWWKNLKMMII 
               
               
                   
               
               
                 LGVICAIILIIIIVYFST. 
               
            
           
         
       
     
     In some embodiments, a protein (e.g., a BoNT E variant) cleaves a SNAP25 protein. In some embodiments, the SNAP25 protein comprises a sequence set forth in SEQ ID NO: 292 
     
       
         
           
               
            
               
                 MAEDADMRNELEEMQRRADQLADESLESTRRMLQLVEESKDAGIRTLVM 
               
               
                   
               
               
                 LDEQGEQLERIEEGMDQINKDMKEAEKNLTDLGKFCGLCVCPCNKLKSS 
               
               
                   
               
               
                 DAYKKAWGNNQDGVVASQPARVVDEREQMAISGGFIRRVTNDARENEMD 
               
               
                   
               
               
                 ENLEQVSGIIGNLRHMALDMGNEIDTQNRQIDRIMEKADSNKTRIDEAN 
               
               
                   
               
               
                 QRATKMLGSG. 
               
            
           
         
       
     
     In some embodiments, a protein (e.g., a BoNT E variant) cleaves a SNAP23 protein. In some embodiments, the SNAP23 protein comprises a sequence set forth in SEQ ID NO: 293 
     
       
         
           
               
            
               
                 MDNLSSEEIQQRAHQITDESLESTRRILGLAIESQDAGIKTITMLDEQK 
               
               
                   
               
               
                 EQLNRIEEGLDQINKDMRETEKTLTELNKCCGLCVCPCNRTKNFESGKA 
               
               
                   
               
               
                 YKTTWGDGGENSPCNVVSKQPGPVTNGQLQQPTTGAASGGYIKRITNDA 
               
               
                   
               
               
                 REDEMEENLTQVGSILGNLKDMALNIGNEIDAQNPQIKRITDKADTNRD 
               
               
                   
               
               
                 RIDIANARAKKLIDS. 
               
            
           
         
       
     
     In some embodiments, a protein (e.g., a BoNT E variant) cleaves a Phosphatase and tensin homolog (PTEN) protein. In some embodiments, the PTEN protein comprises a sequence set forth in SEQ ID NO: 294 
     
       
         
           
               
            
               
                 MTAIIKEIVSRNKRRYQEDGFDLDLTYIYPNIIAMGFPAERLEGVYRNN 
               
               
                   
               
               
                 IDDVVRFLDSKHKNHYKIYNLCAERHYDTAKFNCRVAQYPFEDHNPPQL 
               
               
                   
               
               
                 ELIKPFCEDLDQWLSEDDNHVAAIHCKAGKGRTGVMICAYLLHRGKFLK 
               
               
                   
               
               
                 AQEALDFYGEVRTRDKKGVTIPSQRRYVYYYSYLLKNHLDYRPVALLFH 
               
               
                   
               
               
                 KMMFETIPMFSGGTCNPQFVVCQLKVKIYSSNSGPTRREDKFMYFEFPQ 
               
               
                   
               
               
                 PLPVCGDIKVEFFHKQNKMLKKDKMFHFWVNTFFIPGPEETSEKVENGS 
               
               
                   
               
               
                 LCDQEIDSICSIERADNDKEYLVLTLTKNDLDKANKDKANRYFSPNFKV 
               
               
                   
               
               
                 KLYFTKTVEEPSNPEASSSTSVTPDVSDNEPDHYRYSDTTDSDPENEPF 
               
               
                   
               
               
                 DEDQHTQITKV. 
               
            
           
         
       
     
     In some embodiments, a BoNT E variant protease comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, or at least 30 amino acid mutations set forth in Table 1. 
     In some embodiments, a BoNT F variant protease comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, or at least 30 amino acid mutations set forth in Table 2. 
     In some embodiments, at least one of the amino acid sequence mutations of a BoNT E variant is introduced at an amino acid position selected from the group consisting of 118, C26, Q27, E28, F29 Y68, L89, S99, G101, N118, G127, Q141, E154, E159, N161, S162, S163, R168, M172, K225, C231, 1232, 1233, N238, Q295, Q354, Y357, 1396, P398, L404, and 1409. 
     In some embodiments, a BoNT E variant comprises one or more mutations selected from Q27H, S99A, G101S, N118D, E159L, N161Y, S162Q, S163R, M172K, I132T, Q354R, Y357P. 
     In some embodiments, at least one of the amino acid mutations of a BoNT E variant is selected from the group consisting of 118V, C26Y, Q27H, E28K, F29L, Y68H, L89P, S99A, S99T, G101S, N118D, G127S, Q141K, E154G, E159L, N161Y, S162Q, S163R, R168K, M172K, K225E, C231R, I232T, I233T, N238S, Q295R, I396S, P398L Q354R, Y357P, L404*, and 1409T. In some embodiments, a BoNT E variant (e.g., BoNT L2B from Table 7) comprises the following mutations Q27H, S99A, G101S, N118D, E159L, N161Y, S162Q, S163R, M172K, I132T, Q354R, and Y357P. 
     In some embodiments, at least one of the amino acid sequence mutations of a BoNT F variant is introduced at an amino acid position selected from the group consisting of S70, N76, V106, E164, S166, S167, N184, V193, Y199, E200, S224, R240, A258, N276, L291, T335, S350, F360, Y372, L375, N396, P410, D418, G420, L421, V422, E423, K424, 1425, and V426. 
     In some embodiments, at least one of the amino acid mutations of a BoNT F variant is selected from the group consisting of 570F, N76D, V106A, E164K, S166Y, S1671, N184K, V193M, Y199H, E200G, E200K, S224I, R240F, R240L, A258S, N276S, N276T, L291M, T335S, S350G, F360L, Y372H, L375R, N396H, P410L, D418Y, G420A, L421W, V422L, E423R, K424, I425S, V426*. 
     In some embodiments, a BoNT F variant (e.g., BoNT F 2020-L2A; SEQ ID NO.: 390) comprises the following mutations: V106A, S166Y, S1671, E200G, S224I, R240L, S350G, F360L, Y372H, P410L, G420A, L421W, V422L, E423R, I425S, and V426*. In some embodiments, a BoNT F variant (e.g., BoNT F 2020-L3A; SEQ ID NO.: 391) comprises the following mutations: S166Y, N184K, E200G, S224I, R240F, T335S, F360L Y372H, N396H, P410L, D418Y, and E423K. 
     In some embodiments, at least one of the amino acid mutations of a BoNT F variant is selected from the group consisting of K29, K31, Y72, N99, V106, Y113, V131, S141, 1150, V155, S166, S167, M174, G177, G178, N184, V193, E200, Y210, T214, E215, S224, R240, F267, F270, N293, 1297, R303, T335, S350, F360, Y372, N396, P410, D418, F420, and E423. 
     In some embodiments, at least one of the amino acid mutations of a BoNT F variant (e.g., a BoNT F variant described in Table 29) is selected from the group consisting of K29E, K31N, Y72H, N99S, V106A, Y113C, V131G, S141T, I150T, V1551, S166Y, S1671, M174T, G177A, G178A, N184T, V193M, E200G, Y210H, T214I, E215G, S224I, R240L, F267I, F270V, N293D, I297L, R303C, T335S, S350G, F360L, Y372H, N396H, P410L, D418Y, F420S, and E423K. 
     In some embodiments, a BoNT F variant (e.g., BoNT F 3041-L2D; SEQ ID NO.: 392) comprises the following mutations: K29E, V106A, I150T, S166Y, S1671, M174T, E200G, S224I, R240L, R303C, S350G, F360L, Y372H, N396H, and P410L. In some embodiments, a BoNT F variant (e.g., BoNT 3041-L2F; SEQ ID NO.: 393) comprises the following mutations: Y72H, V106A, V131G, S141T, S166Y, S1671, M174T, E200G, S224I, R240L, S350G, F360L, Y372H, N396H, and P410L. 
     In some embodiments, at least one of the amino acid mutations of a BoNT F variant is selected from the group consisting of N6, Y10, R49, 152, D58, E60, A63, E66, S70, T90, V106, T123, T132, V145, G159, D161, S166, S167, N184, E200, Y201, N211, F217, S224, A226, A232, R240, 1262, L264, D274, N314, G325, D331, S333, T335, N339, S350, F360, T367, F369, Y372, V377, N379, N396, N409, P410, D418, and E423. 
     In some embodiments, at least one of the amino acid mutations of a BoNT F variant (e.g., a BoNT F variant described in Table 30) is selected from the group consisting of N6S, Y10C, R49L, I52V, D58Y, E60D, A63V, E66K, S70H, T901, V106A, T123M, T123S, T1321, V1451, G159S, D161G, S166Y, S1671, N184K, E200G, Y201H, N211S, F217L, S224I, A226S, A232T, R240L, I262T, L264M, D274M, N314S, G325S, D331G, S333F, T335I, N339S, S350G, F360L, T367S, F369F, Y372H, V377I, N379H, N396H, N409D, P410L, D418Y, and E423K. 
     In some embodiments, a BoNT E variant comprises the amino acid sequence as set forth in any one of SEQ ID NOs.: 1-100. In some embodiments, a BoNT F variant comprises the amino acid sequence as set forth in any one of SEQ ID NOs.: 101-285. In some embodiments, a BoNT F variant comprises the amino acid sequence as set forth in any one of SEQ ID NOs.: 390-393. 
     In some embodiments, an evolved BoNT protease variant comprises a BoNT heavy chain. Generally, a BoNT heavy chain comprises a neurotoxin HCC domain, and a neurotoxin translocation domain (HCN). Without wishing to be bound by any particular theory, the HCC domain binds to specific receptors from the presynaptic terminal of a cell, and the HCN domain translocates the BoNT LC into the cell, resulting in intracellular delivery of the catalytic domain of the protease. In some embodiments, an evolved BoNT protease variant (e.g., a BoNT E variant or a BoNT F variant) further comprises a neurotoxin HCC domain, and/or a neurotoxin translocation domain (HCN), for example, a Botulinum toxin HCC or HCN domain or a Tetanus toxin HCC or HCN domain. 
     
       
         
           
               
            
               
                 SEQ ID NO: 295 
               
               
                 CKSVIPRKGTKAPPRLCIRVNNRELFFVASESSYNENDINTPKEIDDTTN 
               
               
                   
               
               
                 LNNNYRNNLDEVILDYNSETIPQISNQTLNTLVQDDSYVPRYDSNGTSEI 
               
               
                   
               
               
                 EEHNVVDLNVFFYLHAQKVPEGETNISLTSSIDTALSEESQVYTFFSSEF 
               
               
                   
               
               
                 INTINKPVHAALFISWINQVIRDFTTEATQKSTFDKIADISLVVPYVGLA 
               
               
                   
               
               
                 LNIGNEVQKENFKEAFELLGAGILLEFVPELLIPTILVFTIKSFIGSSEN 
               
               
                   
               
               
                 KNKIIKAINNSLMERETKWKEIYSWIVSNWLTRINTQFNKRKEQMYQALQ 
               
               
                   
               
               
                 NQVDAIKTVIEYKYNNYTSDERNRLESEYNINNIREELNKKVSLAMENIE 
               
               
                   
               
               
                 RFITESSIFYLMKLINEAKVSKLREYDEGVKEYLLDYISEHRSILGNSVQ 
               
               
                   
               
               
                 ELNDLVTSTLNNSIPFELSSYTNDKILILYF (BoNT F HC N , 
               
               
                   
               
               
                 translocation domain); 
               
               
                   
               
               
                 SEQ ID NO: 296 
               
               
                 NKLYKKIKDNSILDMRYENNKFIDISGYGSNISINGDVYIYSTNRNQFGI 
               
               
                   
               
               
                 YSSKPSEVNIAQNNDIIYNGRYQNFSISFWVRIPKYFNKVNLNNEYTIID 
               
               
                   
               
               
                 CIRNNNSGWKISLNYNKIIWTLQDTAGNNQKLVFNYTQMISISDYINKWI 
               
               
                   
               
               
                 FVTITNNRLGNSRIYINGNLIDEKSISNLGDIHVSDNILFKIVGCNDTRY 
               
               
                   
               
               
                 VGIRYFKVFDTELGKTEIETLYSDEPDPSILKDFWGNYLLYNKRYYLLNL 
               
               
                   
               
               
                 LRTDKSITQNSNFLNINQQRGVYQKPNIFSNTRLYTGVEVIIRKNGSTDI 
               
               
                   
               
               
                 SNTDNFVRKNDLAYINVVDRDVEYRLYADISIAKPEKIIKLIRTSNSNNS 
               
               
                   
               
               
                 LGQIIVMDSIGNNCTMNFQNNNGGNIGLLGFHSNNLVASSWYYNNIRKNT 
               
               
                   
               
               
                 SSNGCFWSFISKEHGWQEN (BoNT F HC C , Binding domain);  
               
               
                   
               
               
                 SEQ ID NO: 297 
               
               
                 CKNIVSVKGIRKSICIEINNGELFFVASENSYNDDNINTPKEIDDTVTSN 
               
               
                   
               
               
                 NNYENDLDQVILNFNSESAPGLSDEKLNLTIQNDAYIPKYDSNGTSDIEQ 
               
               
                   
               
               
                 HDVNELNVFFYLDAQKVPEGENNVNLTSSIDTALLEQPKIYTFFSSEFIN 
               
               
                   
               
               
                 NVNKPVQAALFVSWIQQVLVDFTTEANQKSTVDKIADISIVVPYIGLALN 
               
               
                   
               
               
                 IGNEAQKGNFKDALELLGAGILLEFEPELLIPTILVFTIKSFLGSSDNKN 
               
               
                   
               
               
                 KVIKAINNALKERDEKWKEVYSFIVSNWMTKINTQFNKRKEQMYQALQNQ 
               
               
                   
               
               
                 VNAIKTIIESKYNSYTLEEKNELTNKYDIKQIENELNQKVSIAMNNIDRF 
               
               
                   
               
               
                 LTESSISYLMKLINEVKINKLREYDENVKTYLLNYIIQHGSILGESQQEL 
               
               
                   
               
               
                 NSMVTDTLNNSIPFKLSSYTDDKILISYFNKFFKRIKS (BoNT E 
               
               
                   
               
               
                 HC N , translocation domain); 
               
               
                   
               
               
                 SEQ ID NO: 298 
               
               
                 SSVLNMRYKNDKYVDTSGYDSNININGDVYKYPTNKNQFEIYNDKLSEVN 
               
               
                   
               
               
                 ISQNDYIIYDNKYKNFSISFWVRIPNYDNKIVNVNNEYTIINCMRDNNSG 
               
               
                   
               
               
                 WKVSLNHNEIIWTLQDNAGINQKLAFNYGNANGISDYINKWIFVTITNDR 
               
               
                   
               
               
                 LGDSKLYINGNLIDQKSILNLGNIHVSDNILFKIVNCSYTRYIGIRYFNI 
               
               
                   
               
               
                 FDKELDETEIQTLYSNEPNTNILKDFWGNYLLYDKEYYLLNVLKPNNFID 
               
               
                   
               
               
                 RRKDSTLSINNIRSTILLANRLYSGIKVKIQRVNNSSTNDNLVRKNDQVY 
               
               
                   
               
               
                 INFVASKTHLFPLYADTATTNKEKTIKISSSGNRFNQVVVMNSVGNNCTM 
               
               
                   
               
               
                 NFKNNNGNNIGLLGFKADTVVASTWYYTHMRDHTNSNGCFWNFISEEHGW 
               
               
                   
               
               
                 QEK (BoNT E HC C , Binding domain) 
               
            
           
         
       
     
     In some aspects, the disclosure provides a pharmaceutical composition comprising a protein (e.g., a BoNT E variant or a BoNT F variant) as described herein and a pharmaceutically acceptable excipient. 
     In some aspects, the disclosure provides an isolated nucleic acid encoding a BoNT E variant or a BoNT F variant comprising an amino acid sequence as set forth in any one of SEQ ID NOs.: 1-285 or 390-393. In some aspects, the isolated nucleic acid is contained in a host cell, for example, a bacterial cell, yeast cell, or mammalian cell (e.g., human cell, mouse cell, etc.). 
     In some aspects, the disclosure provides a method of cleaving an intracellular protein, the method comprising delivering to a cell a BoNT E variant or a BoNT F variant as described herein, wherein the protein contacts the intracellular protein in the cell. In some embodiments, the cell is in a subject, for example, a mammalian subject, such as a human or mouse. 
     In some aspects, the disclosure provides a method for reducing VAMP7 activity in a subject, the method comprising administering to the subject an effective amount of a BoNT E variant or a BoNT F variant as described herein. In some embodiments, the subject has or is suspected of having a disease characterized by increased or aberrant VAMP7 activity, for example, cancer, transplantation rejection, or graft-versus-host disease. 
     In some aspects, the disclosure provides a method for reducing VAMP8 activity in a subject, the method comprising administering to the subject an effective amount of a BoNT E variant or a BoNT F variant as described herein. 
     In some aspects, the disclosure provides a method for reducing PTEN activity in a subject, the method comprising administering to the subject an effective amount of a BoNT E variant as described herein. In some embodiments, the subject has or is suspected of having a disease characterized by neuro-degeneration and in which inhibition of PTEN activity could transiently induce cellular regeneration, for example, ischemic neuronal injury (stroke), Parkinson&#39;s Disease, Huntington&#39;s Disease, Alzheimer&#39;s Disease, or spinal cord injury. 
     In some aspects, the disclosure provides a method for reducing SNAP23 activity in a subject, the method comprising administering to the subject an effective amount of a BoNT E variant as described herein. In some embodiments, the subject has or is suspected of having a disease characterized by hypersecretion and in which cleavage of SNAP23 could prevent said secretion, for example, diabetes, autoimmune disorders or Cushing Disease. 
     In some aspects, the disclosure relates to methods of producing an evolved BoNT E variant or BoNT F variant using phage-assisted continuous evolution (PACE). The general concept of PACE technology been described, for example, in International PCT Application, PCT/US2009/056194, filed Sep. 8, 2009, published as WO 2010/028347 on Mar. 11, 2010; International PCT Application, PCT/US2011/066747, filed Dec. 22, 2011, published as WO 2012/088381 on Jun. 28, 2012; U.S. Application, U.S. Ser. No. 13/922,812, filed Jun. 20, 2013; International PCT Applications, PCT/US2015/057012, filed on Oct. 22, 2015, published as WO 2016/077052; and PCT/US2016/027795, filed on Apr. 15, 2016, published as WO 2016/168631, the entire contents of each of which are incorporated herein by reference. In some embodiments, evolved proteases described herein (e.g., proteases evolved using PACE technology described herein) cleave certain substrates (e.g., VAMP1, VAMP2, VAMP7, VAMPS, SNAP25, SNAP23 or PTEN) with higher efficiency and/or specificity relative to previously described BoNT proteases. 
     The summary above is meant to illustrate and outline, in a non-limiting manner, some of the embodiments, advantages, features, and uses of the technology disclosed herein. The disclosure is, however, not limited to the embodiments described in the summary above. Other embodiments, advantages, features, and uses of the technology disclosed herein will be apparent from the Detailed Description, the Drawings, the Examples, and the Claims. 
    
    
     
       BRIEF DESCRIPTION OF DRAWINGS 
         FIG. 1  is a schematic depicting the mechanism of intoxication by Botulinum neurotoxins. 
         FIG. 2  is a schematic overview of Phage-assisted Continuous Evolution (PACE). 
         FIG. 3  is a schematic depiction of proteolysis-activated T7 RNA polymerase for PACE selection of proteases. 
         FIGS. 4A-4D  show PACE selection for the evolution of BoNT proteases.  FIG. 4A  shows an alignment of VAMP1 and VAMP7 protein sequences. Bold residues have been experimentally determined to be important for BoNT F cleavage activity. Underlined residues are fully conserved.  FIG. 4B  shows representative T7-PAP constructs for incorporation into PACE accessory plasmids (AP&#39;s).  FIG. 4C  shows analysis of relative cleavage activity for a selection of BoNT-expressing phage on VAMP1- and VAMP2-derived T7-PAP constructs in a luciferase reporter assay.  FIG. 4D  shows evolution of novel activity in BoNT F phage clones through iterative PACE. BoNT F(1.5) indicates a representative phage clone isolated after PACE selection on the T7-PAP(VAMP71.5) AP. From top to bottom, sequences correspond to SEQ ID NOs: 324-333. 
         FIG. 5  is a schematic depicting an orthogonal polymerase strategy against unselective proteases, and examples of negative selection constructs. From top to bottom, sequences correspond to SEQ ID NOs: 334-337. 
         FIGS. 6A-6B  show proteolytic activity of BoNT proteases.  FIG. 6A  shows BoNT Light Chain (LC) proteolytic activity in SNARE-derived PA-RNAP constructs.  FIG. 6B  shows comparative data for wild-type BoNT serotypes B and F to PACE evolved clones displaying improved VAMP1 and VAMP2 cleavage activity. 
         FIG. 7  shows a primary sequence alignment of VAMP1, VAMP7, and VAMP8. The BoNT LC cleavage sites for serotypes B and F are marked on the bottom and top, respectively. Residues marked with an arrow lead to a decrease in cleavage activity by at least 50% on VAMP2 upon removal of the residue side chain. From top to bottom, sequences correspond to SEQ ID NOs: 338-341. 
         FIG. 8  shows evolved BoNT F variants displaying varied activity on a collection of VAMP1 single residue mutants. L1/L2 were evolved to cleave VAMP1 L55A, L3/4 were evolved to cleave VAMP1 D58G, and L5/L6 were evolved to cleave VAMP1 D65I. 
         FIG. 9  shows luciferase assay data indicating that BoNT B and BoNT F can be evolved to cleave VAMP1/2. 
         FIG. 10  shows an alignment of the amino acid sequences of VAMP1, VAMP7 and VAMP8 (top) and an example of a VAMP1 to VAMP7 evolutionary trajectory (bottom). AP: accessory plasmid. From top to bottom, sequences correspond to SEQ ID NOs: 342-346. 
         FIG. 11  shows an alignment of BoNT F and BoNT B VAMP2 (a natural substrate) cleavage domains with VAMP7. From top to bottom, sequences correspond to SEQ ID NOs: 347-349. 
         FIG. 12  shows data indicating that evolution of BoNT B and F by PACE (e.g., using AP&#39;s 977, 983 and 986 for BoNT F) resulted in BoNT variants with improved activity. 
         FIG. 13  shows representative data relating to validation of BoNT Light Chain (LC) selection; data indicate that evolution of BoNT F protease on VAMP1 enriches for the S166Y mutation, which confers broadly increased activity. 
         FIG. 14  shows one example of a stepping-stone evolutionary pathway for production of BoNT F variants that cleave VAMP7. 
         FIG. 15  shows protease activity assays for BoNT F variants from three different experiments (Lagoons 1-6). Each experiment produced clones that cleave VAMP1 substrates containing a different single site mutation (L55A, D58G, or D65I). 
         FIG. 16  is a schematic depiction of PACE experiments to evolve BoNT F that cleave double mutant substrates; the amino acid sequence of the double mutant VAMP1 (L55A/D58G) is also shown. From top to bottom, sequences correspond to SEQ ID NOs: 347-349. 
         FIG. 17  shows protease-dependent luciferase assay data indicating that certain BoNT F variants produced by PACE can cleave the double mutant VAMP1 substrate (LSSA/D58G). 
         FIG. 18  shows one example of a VAMP1-VAMP7 stepping stone evolutionary trajectory. From top to bottom, sequences correspond to SEQ ID NOs: 350-351. 
         FIG. 19  shows protease-dependent luciferase assay data indicating that certain BoNT F variants produced by PACE can cleave the triple mutant VAMP1 substrate (L55A/D58G/Q59E). From top to bottom, sequences correspond to SEQ ID NOs: 350-351 
         FIG. 20  shows protease-dependent luciferase assay data indicating that certain BoNT F variants produced by PACE can cleave the triple mutant VAMP1 substrate (L55A/D58G/Q59E/K60R). From top to bottom, sequences correspond to SEQ ID NOs: 350-351. 
         FIG. 21  shows data indicating that the activity of proteases on VAMP1 substrates containing mutations V44K (shown as V43 in the figure) and Q32M (shown as Q33 in the figure) can be readily evolved. From top to bottom, sequences correspond to SEQ ID NOs: 352-353. 
         FIG. 22  shows data indicating that iterative selection on progressively more complex VAMP substrates produces several BoNT F variants that cleave VAMP7. 
         FIG. 23  shows an alignment of VAMP1 and VAMP7 amino acid sequences, along with AP-V7-194KL, which contains seven VAMP7 mutations (V44K/K53L/L55A/D58G/Q59E/K60R/D65I). From top to bottom, sequences correspond to SEQ ID NOs: 354, 350, and 351. 
         FIG. 24  shows protein blot analysis for protein expression of two BoNT F evolved variants (2020 L2A, 2020 L3A). 
         FIG. 25  is a schematic diagram of an alignment of VAMP1 and VAMP8 amino acid sequences and double mutant accessory plasmids (APs). From top to bottom, sequences correspond to SEQ ID NOs: 355-356. 
         FIG. 26  shows representative data that indicates VAMP7-evolved BoNT F proteases have a broadened activity profile. 
         FIG. 27  shows an alignment of VAMP1 and VAMP8 amino acid sequences, along with several APs used to evolve VAMP8-cleaving BoNT F variants. Data indicate that VAMP8 APs have high background, but BoNT F variants that cleave VAMP8 were identified. From top to bottom, sequences correspond to SEQ ID NOs: 355-356. 
         FIG. 28  shows data indicating that wild-type BoNT E cleaves SNAP25 protein. 
         FIG. 29  shows that mutation of the residue at position 179 (e.g., D179K) of SNAP25 abolished protease activity by BoNT E. 
         FIG. 30  shows PACE strategies used to generate SNAP23 cleaving BoNT E variants as well as data indicating that several BoNT E variants produced by PACE can cleave SNAP23. 
         FIG. 31  shows a portion of the SNAP25 amino acid sequence and the peptide bonds at which wild-type BoNT A and BoNT E proteases that cleave SNAP25, but not SNAP23. From top to bottom, sequences correspond to SEQ ID NOs: 357-362. 
         FIG. 32  shows a stepping stone schematic for PACE of BoNT E to cleave the therapeutic target phosphatase and tensin homolog (PTEN). From top to bottom, sequences correspond to SEQ ID NOs: 363-389. 
         FIG. 33  shows luminescence assay data for several BoNT E variants that cleave PTEN. 
         FIG. 34  shows an evolved BoNT E variant (L2F 031017) was expressed and purified by His-tag affinity chromatography and eluted with increasing concentrations of imidazole. 
         FIG. 35  shows data indicating that BoNT E L2F cleaves both SNAP25 and PTEN substrates. 
         FIG. 36  shows data indicating that negative selection PACE results in improved PTEN substrate cleavage by BoNT E variants. 
         FIG. 37  shows data indicating BoNT E variant L2B after both positive and negative selection PACE cleaves full-length human PTEN protein at a single peptide bond yielding fragments of approximately the expected molecular weight. 
         FIGS. 38A-38B  show protease expression and isolation of evolved BoNT proteases.  FIG. 38A  shows a Western blot of evolved BoNT F protease m2020-L2A (“m” indicates a maltose-binding protein tag on the N-terminus of the protein).  FIG. 38B  shows a Western blot of Ni-NTA (top) purified BoNT F proteases m2020-L2A and m2020-L3A and subsequent Amylose-purification of BoNT F proteases m2020-L2A and m2020-L3A.  FIG. 39  shows data indicating that BoNT F variant 2020-L2A protease is active in vitro, as measured by a VAMP7 cleavage assay. SEQ ID NOs: 394-395 (top and bottom) are shown. 
         FIG. 40  shows data indicating that the cleavage site of BoNT F variant 2020-L2A protease in VAMP7 has shifted relative to the predicted cleavage site, as measured by MS. SEQ ID NOs: 396-401 (top and bottom) are shown. 
         FIG. 41  shows results of a high-stringency PACE experiment to improve VAMP7 cleavage activity of BoNT F protease variants (PACE-3401). Sequences for VAMP2 (SEQ ID NO: 396) and VAMP7 (SEQ ID NO: 397) are shown. 
         FIG. 42  shows luciferase assay data comparing 2020-L2A and 2020-L3A BoNT F protease-containing phage to PACE-3041 clones. 122-951-proB=low stringency VAMP1; 122-092-proB-low stringency VAMP7; 314-092QS-proB=high stringency VAMP7. 
         FIG. 43  shows luciferase assay data comparing 2020-L2A and 2020-L3A BoNT F protease-containing phage to PACE-3041 clones isolated from 314-092QS-proB lagoons. 
         FIG. 44  shows data relating to in vitro characterization of BoNT F protease variants (m3041-L2D and m3041-L2F). 
         FIG. 45  shows data relating to in vitro validation of BoNT F variants m3041-L2D and m3041-L2F. Clone m3041-L2F was observed to have retained catalytic activity in vitro. 
         FIG. 46  shows data relating to selectivity of evolved BoNT protease variants. Off-target cleavage was observed for BoNT F 2020-L2A samples. 
         FIG. 47  shows data relating to PACE experiment PACE-2300 (VAMP7 positive selection, VAMP1 negative selection). 313-092-proB (VAMP7+)=positive selection on VAMP7 substrate; T3neg-951-proD (VAMP1−)=simultaneous negative selection on VAMP1 substrate. VAMP2 (SEQ ID NO: 396) and VAMP7 (SEQ ID NO: 397) are shown. 
         FIG. 48  shows luciferase assay data relating to PACE experiment PACE-2300. one clone that possesses apparent selectivity: BoNT F(2300-L3B). 
         FIG. 49  shows data relating to the in vitro characterization of m2300-L3B. m2300-L3B protease retains activity on VAMP7. Improvements in in vitro selectivity for VAMP1/7 for m2300-L3B versus m2020-L2A were observed. Substantial improvements in in vitro selectivity for VAMP1/7 for m2300-L3B versus m2020-L3A were observed. 
     
    
    
     DEFINITIONS 
     The term “protease,” as used herein, refers to an enzyme that catalyzes the hydrolysis of a peptide (amide) bond linking amino acid residues together within a protein. The term embraces both naturally occurring and engineered proteases. Many proteases are known in the art. Proteases can be classified by their catalytic residue, and protease classes include, without limitation, serine proteases (serine alcohol), threonine proteases (threonine secondary alcohol), cysteine proteases (cysteine thiol), aspartate proteases (aspartate carboxylic acid), glutamic acid proteases (glutamate carboxylic acid), and metalloproteases (metal ion, e.g., zinc). The structures in parentheses correlate to the respective catalytic moiety of the proteases of each class. Some proteases are highly promiscuous and cleave a wide range of protein substrates, e.g., trypsin or pepsin. Other proteases are highly specific and only cleave substrates with a specific sequence. Some blood clotting proteases such as, for example, thrombin, and some viral proteases such as, for example, HCV or TEV protease, are highly specific proteases. In another example, Botulinum toxin proteases (BoNTs) generally cleave specific SNARE proteins. Proteases that cleave in a very specific manner typically bind to multiple amino acid residues of their substrate. Suitable proteases and protease cleavage sites, also sometimes referred to as “protease substrates,” will be apparent to those of skill in the art and include, without limitation, proteases listed in the MEROPS database, accessible at merops.sanger.ac.uk and described in Rawlings et al., (2014) MEROPS: the database of proteolytic enzymes, their substrates and inhibitors.  Nucleic Acids Res  42, D503-D509, the entire contents of each of which are incorporated herein by reference. The disclosure is not limited in this respect. 
     The term “protein,” as used herein, refers to a polymer of amino acid residues linked together by peptide bonds. The term, as used herein, refers to proteins, polypeptides, and peptides of any size, structure, or function. Typically, a protein will be at least three amino acids long. A protein may refer to an individual protein or a collection of proteins. Inventive proteins preferably contain only natural amino acids, although non-natural amino acids (i.e., compounds that do not occur in nature but that can be incorporated into a polypeptide chain; see, for example, cco.caltech.edu/˜dadgrp/Unnatstruct.gif, which displays structures of non-natural amino acids that have been successfully incorporated into functional ion channels) and/or amino acid analogs as are known in the art may alternatively be employed. Also, one or more of the amino acids in an inventive protein may be modified, for example, by the addition of a chemical entity such as a carbohydrate group, a hydroxyl group, a phosphate group, a farnesyl group, an isofarnesyl group, a fatty acid group, a linker for conjugation, functionalization, or other modification, etc. A protein may also be a single molecule or may be a multi-molecular complex. A protein may be just a fragment of a naturally occurring protein or peptide. A protein may be naturally occurring, recombinant, or synthetic, or any combination of these. 
     The term “Botulinum neurotoxin (BoNT) protease,” as used herein, refers to a protease derived from, or having at least 70% sequence homology to (or at least 70% identity to) a Botulinum neurotoxin (BoNT), for example, a BoNT derived from a bacterium of the genus  Clostridium  (e.g.,  C. botulinum ). Structurally, BoNT proteins comprise two conserved domains, a “heavy chain” (HC) and a “light chain” (LC). The LC comprises a zinc metalloprotease domain responsible for the catalytic activity of the protein. The HC typically comprises a HCC domain, which is responsible for binding to neuronal cells, and a HCN domain, which mediates translocation of the protein into a cell. Examples of BoNT HC domains are represented by the amino acid sequences set forth in SEQ ID NO: 299 and 300 below. 
                    BoNT E HCC Domain       SSVLNMRYKNDKYVDTSGYDSNININGDVYKYPTNKNQFEIYNDKLSEVN               ISQNDYIIYDNKYKNFSISFWVRIPNYDNKIVNVNNEYTIINCMRDNNSG               WKVSLNHNEIIWTLQDNAGINQKLAFNYGNANGISDYINKWIFVTITNDR               LGDSKLYINGNLIDQKSILNLGNIHVSDNILFKIVNCSYTRYIGIRYFNI               FDKELDETEIQTLYSNEPNTNILKDFWGNYLLYDKEYYLLNVLKPNNFID               RRKDSTLSINNIRSTILLANRLYSGIKVKIQRVNNSSTNDNLVRKNDQVY               INFVASKTHLFPLYADTATTNKEKTIKISSSGNRFNQVVVMNSVGNNCTM               NFKNNNGNNIGLLGFKADTVVASTWYYTHMRDHTNSNGCFWNFISEEHGW               QEK (BoNT E HC C , Binding domain)               BoNT E HCN Domain       CKNIVSVKGIRKSICIEINNGELFFVASENSYNDDNINTPKEIDDTVTSN               NNYENDLDQVILNFNSESAPGLSDEKLNLTIQNDAYIPKYDSNGTSDIEQ               HDVNELNVFFYLDAQKVPEGENNVNLTSSIDTALLEQPKIYTFFSSEFIN               NVNKPVQAALFVSWIQQVLVDFTTEANQKSTVDKIADISIVVPYIGLALN               IGNEAQKGNFKDALELLGAGILLEFEPELLIPTILVFTIKSFLGSSDNKN               KVIKAINNALKERDEKWKEVYSFIVSNWMTKINTQFNKRKEQMYQALQNQ               VNAIKTIIESKYNSYTLEEKNELTNKYDIKQIENELNQKVSIAMNNIDRF               LTESSISYLMKLINEVKINKLREYDENVKTYLLNYIIQHGSILGESQQEL               NSMVTDTLNNSIPFKLSSYTDDKILISYFNKFFKRIKS (BoNT E               HC N , translocation domain)            
There are seven serotypes of BoNTs, denoted BoNT A-G. BoNT serotypes A, C, and E cleave synaptosome-associated protein (SNAP25). BoNT serotype C has also been observed to cleave syntaxin. BoNT serotypes B, D, F, and G cleave vesicle-associated membrane proteins (VAMPs). An example of a SNAP25 substrate that is cleaved by wild-type BoNT proteases (e.g., BoNT E) is represented by the amino acid sequence set forth in SEQ ID NO: 301. An example of a VAMP substrate (e.g., VAMP1) that is cleaved by wild-type BoNT proteases (e.g., BoNT E) is represented by the amino acid sequence set forth in SEQ ID NO: 302.
 
     
       
         
           
               
            
               
                 SNAP25 substrate sequence 
               
               
                 MAEDADMRNELEEMQRRADQLADESLESTRRMLQLVEESKDAGIRTLVML 
               
               
                   
               
               
                 DEQGEQLERIEEGMDQINKDMKEAEKNLTDLGKFCGLCVCPCNKLKSSDA 
               
               
                   
               
               
                 YKKAWGNNQDGVVASQPARVVDEREQMAISGGFIRRVTNDARENEMDENL 
               
               
                   
               
               
                 EQVSGIIGNLRHMALDMGNEIDTQNRQIDRIMEKADSNKTRIDEANQRAT 
               
               
                   
               
               
                 KMLGSG 
               
               
                   
               
               
                 VAMP1 substrate sequence 
               
               
                 MSAPAQPPAEGTEGTAPGGGPPGPPPNMTSNRRLQQTQAQVEEVVDIIRV 
               
               
                   
               
               
                 NVDKVLERDQKLSELDDRADALQAGASQFESSAAKLKRKYWWKNCKMMIM 
               
               
                   
               
               
                 LGAICAIIVVVIVRRG 
               
            
           
         
       
     
     A wild-type BoNT protease refers to the amino acid sequence of a BoNT protease as it naturally occurs in a  Clostridium botulinum  genome. Examples of wild-type BoNT proteases are represented by the amino acid sequences set forth in SEQ ID NOs.: 303-309. 
     
       
         
           
               
            
               
                 Botulinum neurotoxin serotype A 
               
               
                 MPFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDT 
               
               
                   
               
               
                 FTNPEEGDLNPPPEAKQVPVSYYDSTYLSTDNEKDNYLKGVTKLFERIYS 
               
               
                   
               
               
                 TDLGRMLLTSIVRGIPFWGGSTIDTELKVIDTNCINVIQPDGSYRSEELN 
               
               
                   
               
               
                 LVIIGPSADIIQFECKSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESL 
               
               
                   
               
               
                 EVDTNPLLGAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAY 
               
               
                   
               
               
                 YEMSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKA 
               
               
                   
               
               
                 KSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYT 
               
               
                   
               
               
                 EDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAAN 
               
               
                   
               
               
                 FNGQNTEINNMNFTKLKNFTGLFEFYKLL 
               
               
                   
               
               
                 Botulinum neurotoxin serotype B 
               
               
                 MPVTINNFNYNDPIDNNNIIMMEPPFARGTGRYYKAFKITDRIWIIPERY 
               
               
                   
               
               
                 TFGYKPEDFNKSSGIFNRDVCEYYDPDYLNTNDKKNIFLQTMIKLFNRIK 
               
               
                   
               
               
                 SKPLGEKLLEMIINGIPYLGDRRVPLEEFNTNIASVTVNKLISNPGEVER 
               
               
                   
               
               
                 KKGIFANLIIFGPGPVLNENETIDIGIQNHFASREGFGGIMQMKFCPEYV 
               
               
                   
               
               
                 SVFNNVQENKGASIFNRRGYFSDPALILMHELIHVLHGLYGIKVDDLPIV 
               
               
                   
               
               
                 PNEKKFFMQSTDAIQAEELYTFGGQDPSIITPSTDKSIYDKVLQNFRGIV 
               
               
                   
               
               
                 DRLNKVLVCISDPNININIYKNKFKDKYKFVEDSEGKYSIDVESFDKLYK 
               
               
                   
               
               
                 SLMFGFTETNIAENYKIKTRASYFSDSLPPVKIKNLLDNEIYTIEEGFNI 
               
               
                   
               
               
                 SDKDMEKEYRGQNKAINKQAYEEISKEHLAVYKIQM 
               
               
                   
               
               
                 Botulinum neurotoxin serotype C 
               
               
                 MPITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRF 
               
               
                   
               
               
                 SRNSNPNLNKPPRVTSPKSGYYDPNYLSTDSDKDTFLKEIIKLFKRINSR 
               
               
                   
               
               
                 EIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVKTRQGNNWVKTG 
               
               
                   
               
               
                 SINPSVIITGPRENIIDPETSTFKLTNNTFAAQEGFGALSIISISPRFML 
               
               
                   
               
               
                 TYSNATNDVGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTIS 
               
               
                   
               
               
                 SVTSNIFYSQYNVKLEYAEIYAFGGPTIDLIPKSARKYFEEKALDYYRSI 
               
               
                   
               
               
                 AKRLNSITTANPSSFNKYIGEYKQKLIRKYRFVVESSGEVTVNRNKFVEL 
               
               
                   
               
               
                 YNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFN 
               
               
                   
               
               
                 IPKSNLNVLFMGQNLSRNPALRKVNPENMLYLFTKF 
               
               
                   
               
               
                 Botulinum neurotoxin serotype D 
               
               
                 MTWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERF 
               
               
                   
               
               
                 SSDTNPSLSKPPRPTSKYQSYYDPSYLSTDEQKDTFLKGIIKLFKRINER 
               
               
                   
               
               
                 DIGKKLINYLVVGSPFMGDSSTPEDTFDFTRHTTNIAVEKFENGSWKVTN 
               
               
                   
               
               
                 IITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLL 
               
               
                   
               
               
                 TFSDVTSNQSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIR 
               
               
                   
               
               
                 PQVSEGFFSQDGPNVQFEELYTFGGLDVEIIPQIERSQLREKALGHYKDI 
               
               
                   
               
               
                 AKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFVVNIDKFNSL 
               
               
                   
               
               
                 YSDLTNVMSEVVYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFN 
               
               
                   
               
               
                 LTNKGFNIENSGQNIERNPALQKLSSESVVDLFTKV 
               
               
                   
               
               
                 Botulinum neurotoxin serotype E 
               
               
                 MTWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERF 
               
               
                   
               
               
                 SSDTNPSLSKPPRPTSKYQSYYDPSYLSTDEQKDTFLKGIIKLFKRINER 
               
               
                   
               
               
                 DIGKKLINYLVVGSPFMGDSSTPEDTFDFTRHTTNIAVEKFENGSWKVTN 
               
               
                   
               
               
                 IITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLL 
               
               
                   
               
               
                 TFSDVTSNQSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIR 
               
               
                   
               
               
                 PQVSEGFFSQDGPNVQFEELYTFGGLDVEIIPQIERSQLREKALGHYKDI 
               
               
                   
               
               
                 AKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFVVNIDKFNSL 
               
               
                   
               
               
                 YSDLTNVMSEVVYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFN 
               
               
                   
               
               
                 LTNKGFNIENSGQNIERNPALQKLSSESVVDLFTKV 
               
               
                   
               
               
                 Botulinum neurotoxin serotype F 
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPERN 
               
               
                   
               
               
                 TIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINS 
               
               
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSII 
               
               
                   
               
               
                 LNLLVLGAGPDIFENSSYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYE 
               
               
                   
               
               
                 YTFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKV 
               
               
                   
               
               
                 KQAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATR 
               
               
                   
               
               
                 LSRVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
               
               
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
               
               
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
               
               
                   
               
               
                 Botulinum neurotoxin serotype G 
               
               
                 MPVNIKNFNYNDPINNDDIIMMEPFNDPGPGTYYKAFRIIDRIWIVPERF 
               
               
                   
               
               
                 TYGFQPDQFNASTGVFSKDVYEYYDPTYLKTDAEKDKFLKTMIKLFNRIN 
               
               
                   
               
               
                 SKPSGQRLLDMIVDAIPYLGNASTPPDKFAANVANVSINKKIIQPGAEDQ 
               
               
                   
               
               
                 IKGLMTNLIIFGPGPVLSDNFTDSMIMNGHSPISEGFGARMMIRFCPSCL 
               
               
                   
               
               
                 NVFNNVQENKDTSIFSRRAYFADPALTLMHELIHVLHGLYGIKISNLPIT 
               
               
                   
               
               
                 PNTKEFFMQHSDPVQAEELYTFGGHDPSVISPSTDMNIYNKALQNFQDIA 
               
               
                   
               
               
                 NRLNIVSSAQGSGIDISLYKQIYKNKYDFVEDPNGKYSVDKDKFDKLYKA 
               
               
                   
               
               
                 LMFGFTETNLAGEYGIKTRYSYFSEYLPPIKTEKLLDNTIYTQNEGFNIA 
               
               
                   
               
               
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     The term “BoNT protease variant,” as used herein, refers to a protein (e.g., a BoNT protease) having one or more amino acid variations introduced into the amino acid sequence, e.g., as a result of application of the PACE method or by genetic engineering (e.g., recombinant gene expression, gene synthesis, etc.), as compared to the amino acid sequence of a naturally-occurring or wild-type BoNT protein (e.g., SEQ ID NO: 286 or SEQ ID NO: 287). Amino acid sequence variations may include one or more mutated residues within the amino acid sequence of the protease, e.g., as a result of a change in the nucleotide sequence encoding the protease that results in a change in the codon at any particular position in the coding sequence, the deletion of one or more amino acids (e.g., a truncated protein), the insertion of one or more amino acids, or any combination of the foregoing. In certain embodiments, a BoNT protease variant cleaves a different target peptide (e.g., has broadened or different substrate specificity) relative to a wild-type BoNT protease. For example, in some embodiments, a BoNT F protease variant cleaves a VAMP7 protein or peptide. 
     The term “continuous evolution,” as used herein, refers to an evolution procedure, in which a population of nucleic acids is subjected to multiple rounds of (a) replication, (b) mutation (or modification of the primary sequence of nucleotides of the nucleic acids in the population), and (c) selection to produce a desired evolved product, for example, a novel nucleic acid encoding a novel protein with a desired activity, wherein the multiple rounds of replication, mutation, and selection can be performed without investigator interaction, and wherein the processes (a)-(c) can be carried out simultaneously. Typically, the evolution procedure is carried out in vitro, for example, using cells in culture as host cells. In general, a continuous evolution process provided herein relies on a system in which a gene of interest is provided in a nucleic acid vector that undergoes a life-cycle including replication in a host cell and transfer to another host cell, wherein a critical component of the life-cycle is deactivated and reactivation of the component is dependent upon a desired variation in an amino acid sequence of a protein encoded by the gene of interest. 
     In some embodiments, the gene of interest (e.g., a gene encoding a BoNT protease) is transferred from cell to cell in a manner dependent on the activity of the gene of interest. In some embodiments, the transfer vector is a virus infecting cells, for example, a bacteriophage or a retroviral vector. In some embodiments, the viral vector is a phage vector infecting bacterial host cells. In some embodiments, the transfer vector is a conjugative plasmid transferred from a donor bacterial cell to a recipient bacterial cell. 
     In some embodiments, the nucleic acid vector comprising the gene of interest is a phage, a viral vector, or naked DNA (e.g., a mobilization plasmid). In some embodiments, transfer of the gene of interest from cell to cell is via infection, transfection, transduction, conjugation, or uptake of naked DNA, and efficiency of cell-to-cell transfer (e.g., transfer rate) is dependent on an activity of a product encoded by the gene of interest. For example, in some embodiments, the nucleic acid vector is a phage harboring the gene of interest and the efficiency of phage transfer (via infection) is dependent on an activity of the gene of interest in that a protein required for the generation of phage particles (e.g., pIII for M13 phage) is expressed in the host cells only in the presence of the desired activity of the gene of interest. 
     For example, some embodiments provide a continuous evolution system, in which a population of viral vectors comprising a gene of interest to be evolved replicates in a flow of host cells, e.g., a flow through a lagoon, wherein the viral vectors are deficient in a gene encoding a protein that is essential for the generation of infectious viral particles, and wherein that gene is in the host cell under the control of a conditional promoter that can be activated by a gene product encoded by the gene of interest, or a mutated version thereof. In some embodiments, the activity of the conditional promoter depends on a desired function of a gene product encoded by the gene of interest. Viral vectors, in which the gene of interest has not acquired a desired function as a result of a variation of amino acids introduced into the gene product protein sequence, will not activate the conditional promoter, or may only achieve minimal activation, while any mutations introduced into the gene of interest that confers the desired function will result in activation of the conditional promoter. Since the conditional promoter controls an essential protein for the viral life cycle, e.g., pIII, activation of this promoter directly corresponds to an advantage in viral spread and replication for those vectors that have acquired an advantageous mutation. 
     The term “flow,” as used herein in the context of host cells, refers to a stream of host cells, wherein fresh host cells are being introduced into a host cell population, for example, a host cell population in a lagoon, remain within the population for a limited time, and are then removed from the host cell population. In a simple form, a host cell flow may be a flow through a tube, or a channel, for example, at a controlled rate. In other embodiments, a flow of host cells is directed through a lagoon that holds a volume of cell culture media and comprises an inflow and an outflow. The introduction of fresh host cells may be continuous or intermittent and removal may be passive, e.g., by overflow, or active, e.g., by active siphoning or pumping. Removal further may be random, for example, if a stirred suspension culture of host cells is provided, removed liquid culture media will contain freshly introduced host cells as well as cells that have been a member of the host cell population within the lagoon for some time. Even though, in theory, a cell could escape removal from the lagoon indefinitely, the average host cell will remain only for a limited period of time within the lagoon, which is determined mainly by the flow rate of the culture media (and suspended cells) through the lagoon. 
     Since the viral vectors replicate in a flow of host cells, in which fresh, uninfected host cells are provided while infected cells are removed, multiple consecutive viral life cycles can occur without investigator interaction, which allows for the accumulation of multiple advantageous mutations in a single evolution experiment. 
     The term “phage-assisted continuous evolution (PACE),” as used herein, refers to continuous evolution that employs phage as viral vectors. 
     The term “viral vector,” as used herein, refers to a nucleic acid comprising a viral genome that, when introduced into a suitable host cell, can be replicated and packaged into viral particles able to transfer the viral genome into another host cell. The term viral vector extends to vectors comprising truncated or partial viral genomes. For example, in some embodiments, a viral vector is provided that lacks a gene encoding a protein essential for the generation of infectious viral particles. In suitable host cells, for example, host cells comprising the lacking gene under the control of a conditional promoter, however, such truncated viral vectors can replicate and generate viral particles able to transfer the truncated viral genome into another host cell. In some embodiments, the viral vector is a phage, for example, a filamentous phage (e.g., an M13 phage). In some embodiments, a viral vector, for example, a phage vector, is provided that comprises a gene of interest to be evolved. 
     The term “nucleic acid,” as used herein, refers to a polymer of nucleotides. The polymer may include natural nucleosides (i.e., adenosine, thymidine, guano sine, cytidine, uridine, deoxyadenosine, deoxythymidine, deoxyguanosine, and deoxycytidine), nucleoside analogs (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5-methylcytidine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, 4-acetylcytidine, 5-(carboxyhydroxymethyl)uridine, dihydrouridine, methylpseudouridine, 1-methyl adenosine, 1-methyl guanosine, N6-methyl adenosine, and 2-thiocytidine), chemically modified bases, biologically modified bases (e.g., methylated bases), intercalated bases, modified sugars (e.g., 2′-fluororibose, ribose, 2′-deoxyribose, 2′-O-methylcytidine, arabinose, and hexose), or modified phosphate groups (e.g., phosphorothioates and 5′-N-phosphoramidite linkages). 
     The term “gene of interest” or “gene encoding a protease of interest,” as used herein, refers to a nucleic acid construct comprising a nucleotide sequence encoding a gene product, e.g., a BoNT protease, of interest to be evolved in a continuous evolution process as described herein. The term includes any variations of a gene of interest that are the result of a continuous evolution process according to methods described herein. For example, in some embodiments, a gene of interest is a nucleic acid construct comprising a nucleotide sequence encoding a protease to be evolved, cloned into a viral vector, for example, a phage genome, so that the expression of the encoding sequence is under the control of one or more promoters in the viral genome. In other embodiments, a gene of interest is a nucleic acid construct comprising a nucleotide sequence encoding a protease to be evolved and a promoter operably linked to the encoding sequence. When cloned into a viral vector, for example, a phage genome, the expression of the encoding sequence of such genes of interest is under the control of the heterologous promoter and, in some embodiments, may also be influenced by one or more promoters in the viral genome. 
     The term “function of a gene of interest,” as interchangeably used with the term “activity of a gene of interest,” refers to a function or activity of a gene product, for example, a nucleic acid or a protein, encoded by the gene of interest. For example, a function of a gene of interest may be an enzymatic activity (e.g., proteolytic activity resulting in the generation of cleavage of a desired substrate, etc.), an ability to activate transcription (e.g., transcriptional activation activity targeted to a specific promoter sequence), a bond-forming activity (e.g., an enzymatic activity resulting in the formation of a covalent bond), or a binding activity (e.g., a protein, DNA, or RNA binding activity). 
     The term “promoter” refers to a nucleic acid molecule with a sequence recognized by the cellular transcription machinery and able to initiate transcription of a downstream gene. A promoter can be constitutively active, meaning that the promoter is always active in a given cellular context, or conditionally active, meaning that the promoter is only active under specific conditions. For example, a conditional promoter may only be active in the presence of a specific protein that connects a protein associated with a regulatory element in the promoter to the basic transcriptional machinery, or only in the absence of an inhibitory molecule. A subclass of conditionally active promoters are inducible promoters that require the presence of a small molecule “inducer” for activity. Examples of inducible promoters include, but are not limited to, arabinose-inducible promoters, Tet-on promoters, and tamoxifen-inducible promoters. A variety of constitutive, conditional, and inducible promoters are well known to the skilled artisan, and the skilled artisan will be able to ascertain a variety of such promoters useful in carrying out the instant invention, which is not limited in this respect. 
     The term “viral particle,” as used herein, refers to a viral genome, for example, a DNA or RNA genome, that is associated with a coat of a viral protein or proteins, and, in some cases, with an envelope of lipids. For example, a phage particle comprises a phage genome packaged into a protein encoded by the wild type phage genome. 
     The term “infectious viral particle,” as used herein, refers to a viral particle able to transport the viral genome it comprises into a suitable host cell. Not all viral particles are able to transfer the viral genome to a suitable host cell. Particles unable to accomplish this are referred to as non-infectious viral particles. In some embodiments, a viral particle comprises a plurality of different coat proteins, wherein one or some of the coat proteins can be omitted without compromising the structure of the viral particle. In some embodiments, a viral particle is provided in which at least one coat protein cannot be omitted without the loss of infectivity. If a viral particle lacks a protein that confers infectivity, the viral particle is not infectious. For example, an M13 phage particle that comprises a phage genome packaged in a coat of phage proteins (e.g., pVIII) but lacks pIII (protein III) is a non-infectious M13 phage particle because pIII is essential for the infectious properties of M13 phage particles. 
     The term “viral life cycle,” as used herein, refers to the viral reproduction cycle comprising insertion of the viral genome into a host cell, replication of the viral genome in the host cell, and packaging of a replication product of the viral genome into a viral particle by the host cell. 
     In some embodiments, the viral vector provided is a phage. The term “phage,” as used herein interchangeably with the term “bacteriophage,” refers to a virus that infects bacterial cells. Typically, phages consist of an outer protein capsid enclosing genetic material. The genetic material can be ssRNA, dsRNA, ssDNA, or dsDNA, in either linear or circular form. Phages and phage vectors are well known to those of skill in the art and non-limiting examples of phages that are useful for carrying out the methods provided herein are λ (Lysogen), T2, T4, T7, T12, R17, M13, MS2, G4, P1, P2, P4, Phi X174, N4, Φ6, and Φ29. In certain embodiments, the phage utilized in the present invention is M13. Additional suitable phages and host cells will be apparent to those of skill in the art, and the invention is not limited in this aspect. For an exemplary description of additional suitable phages and host cells, see Elizabeth Kutter and Alexander Sulakvelidze:  Bacteriophages: Biology and Applications . CRC Press; 1 st  edition (December 2004), ISBN: 0849313368; Martha R. J. Clokie and Andrew M. Kropinski:  Bacteriophages: Methods and Protocols, Volume  1:  Isolation, Characterization, and Interactions  ( Methods in Molecular Biology ) Humana Press; 1 st  edition (December, 2008), ISBN: 1588296822; Martha R. J. Clokie and Andrew M. Kropinski:  Bacteriophages: Methods and Protocols, Volume  2 : Molecular and Applied Aspects  ( Methods in Molecular Biology ) Humana Press; 1 st  edition (December 2008), ISBN: 1603275649; all of which are incorporated herein in their entirety by reference for disclosure of suitable phages and host cells as well as methods and protocols for isolation, culture, and manipulation of such phages). 
     In some embodiments, the phage is a filamentous phage. In some embodiments, the phage is an M13 phage. M13 phages are well known to those in the art and the biology of M13 phages has extensively been studied. Wild type M13 phage particles comprise a circular, single-stranded genome of approximately 6.4 kb. In certain embodiments, the wild-type genome of an M13 phage includes eleven genes, gI-gXI, which, in turn, encode the eleven M13 proteins, pI-pXI, respectively. gVIII encodes pVIII, also often referred to as the major structural protein of the phage particles, while gIII encodes pIII, also referred to as the minor coat protein, which is required for infectivity of M13 phage particles. 
     The M13 life cycle includes attachment of the phage to the sex pilus of a suitable bacterial host cell via the pIII protein and insertion of the phage genome into the host cell. The circular, single-stranded phage genome is then converted to a circular, double-stranded DNA, also termed the replicative form (RF), from which phage gene transcription is initiated. The wild type M13 genome comprises nine promoters and two transcriptional terminators as well as an origin of replication. This series of promoters provides a gradient of transcription such that the genes nearest the two transcriptional terminators (gVIII and IV) are transcribed at the highest levels. In wild-type M13 phage, transcription of all 11 genes proceeds in the same direction. One of the phage-encoded proteins, pII, initiates the generation of linear, single-stranded phage genomes in the host cells, which are subsequently circularized, and bound and stabilized by pV. The circularized, single-stranded M13 genomes are then bound by pVIII, while pV is stripped off the genome, which initiates the packaging process. At the end of the packaging process, multiple copies of pIII are attached to wild-type M13 particles, thus generating infectious phage ready to infect another host cell and concluding the life cycle. 
     The M13 phage genome can be manipulated, for example, by deleting one or more of the wild type genes, and/or inserting a heterologous nucleic acid construct into the genome. M13 does not have stringent genome size restrictions, and insertions of up to 42 kb have been reported. This allows M13 phage vectors to be used in continuous evolution experiments to evolve genes of interest without imposing a limitation on the length of the gene to be involved. 
     The term “selection phage,” as used herein interchangeably with the term “selection plasmid,” refers to a modified phage that comprises a gene of interest to be evolved and lacks a full-length gene encoding a protein required for the generation of infectious phage particles. For example, some M13 selection phages provided herein comprise a nucleic acid sequence encoding a protease to be evolved, e.g., under the control of an M13 promoter, and lack all or part of a phage gene encoding a protein required for the generation of infectious phage particles, e.g., gI, gII, gIII, gIV, gV, gVI, gVII, gVIII, gIX, or gX, or any combination thereof. For example, some M13 selection phages provided herein comprise a nucleic acid sequence encoding a protease to be evolved, e.g., under the control of an M13 promoter, and lack all or part of a gene encoding a protein required for the generation of infective phage particles, e.g., the gIII gene encoding the pIII protein. 
     The term “helper phage,” as used herein interchangeable with the terms “helper phagemid” and “helper plasmid,” refers to a nucleic acid construct comprising a phage gene required for the phage life cycle, or a plurality of such genes, but lacking a structural element required for genome packaging into a phage particle. For example, a helper phage may provide a wild-type phage genome lacking a phage origin of replication. In some embodiments, a helper phage is provided that comprises a gene required for the generation of phage particles, but lacks a gene required for the generation of infectious particles, for example, a full-length pIII gene. In some embodiments, the helper phage provides only some, but not all, genes required for the generation of phage particles. Helper phages are useful to allow modified phages that lack a gene required for the generation of phage particles to complete the phage life cycle in a host cell. Typically, a helper phage will comprise the genes required for the generation of phage particles that are lacking in the phage genome, thus complementing the phage genome. In the continuous evolution context, the helper phage typically complements the selection phage, but both lack a phage gene required for the production of infectious phage particles. 
     The term “replication product,” as used herein, refers to a nucleic acid that is the result of viral genome replication by a host cell. This includes any viral genomes synthesized by the host cell from a viral genome inserted into the host cell. The term includes non-mutated as well as mutated replication products. 
     The term “accessory plasmid,” as used herein, refers to a plasmid comprising a gene required for the generation of infectious viral particles under the control of a conditional promoter. In the context of continuous evolution described herein, the conditional promoter of the accessory plasmid is typically activated by a function of the gene of interest to be evolved. Accordingly, the accessory plasmid serves the function of conveying a competitive advantage to those viral vectors in a given population of viral vectors that carry a gene of interest able to activate the conditional promoter. Only viral vectors carrying an “activating” gene of interest will be able to induce expression of the gene required to generate infectious viral particles in the host cell, and, thus, allow for packaging and propagation of the viral genome in the flow of host cells. Vectors carrying non-activating versions of the gene of interest, on the other hand, will not induce expression of the gene required to generate infectious viral vectors, and, thus, will not be packaged into viral particles that can infect fresh host cells. 
     In some embodiments, the conditional promoter of the accessory plasmid is a promoter the transcriptional activity of which can be regulated over a wide range, for example, over 2, 3, 4, 5, 6, 7, 8, 9, or 10 orders of magnitude by the activating function, for example, function of a protein encoded by the gene of interest). In some embodiments, the level of transcriptional activity of the conditional promoter depends directly on the desired function of the gene of interest. This allows for starting a continuous evolution process with a viral vector population comprising versions of the gene of interest that only show minimal activation of the conditional promoter. In the process of continuous evolution, any mutation in the gene of interest that increases activity of the conditional promoter directly translates into higher expression levels of the gene required for the generation of infectious viral particles, and, thus, into a competitive advantage over other viral vectors carrying minimally active or loss-of-function versions of the gene of interest. 
     The stringency of selective pressure imposed by the accessory plasmid in a continuous evolution procedure as provided herein can be modulated. In some embodiments, the use of low copy number accessory plasmids results in an elevated stringency of selection for versions of the gene of interest that activate the conditional promoter on the accessory plasmid, while the use of high copy number accessory plasmids results in a lower stringency of selection. The terms “high copy number plasmid” and “low copy number plasmid” are art-recognized and those of skill in the art will be able to ascertain whether a given plasmid is a high or low copy number plasmid. In some embodiments, a low copy number accessory plasmid is a plasmid exhibiting an average copy number of plasmid per host cell in a host cell population of about 5 to about 100. In some embodiments, a very low copy number accessory plasmid is a plasmid exhibiting an average copy number of plasmid per host cell in a host cell population of about 1 to about 10. In some embodiments, a very low copy number accessory plasmid is a single-copy per cell plasmid. In some embodiments, a high copy number accessory plasmid is a plasmid exhibiting an average copy number of plasmid per host cell in a host cell population of about 100 to about 5000. The copy number of an accessory plasmid will depend to a large part on the origin of replication employed. Those of skill in the art will be able to determine suitable origins of replication in order to achieve a desired copy number. 
     In some embodiments, the stringency of selective pressure imposed by the accessory plasmid can also be modulated through the use of mutant or alternative conditional transcription factors with higher or lower transcriptional output (e.g., a T7 RNA polymerase comprising a Q649S mutation). In some embodiments, the use of lower transcriptional output results in an elevated stringency of selection for versions of the gene of interest that activate the conditional promoter on the accessory plasmid, while the use of higher transcriptional output machinery results in a lower stringency of selection. 
     It should be understood that the function of the accessory plasmid, namely to provide a gene required for the generation of viral particles under the control of a conditional promoter the activity of which depends on a function of the gene of interest, can be conferred to a host cell in alternative ways. Such alternatives include, but are not limited to, permanent insertion of a gene construct comprising the conditional promoter and the respective gene into the genome of the host cell, or introducing it into the host cell using an different vector, for example, a phagemid, a cosmid, a phage, a virus, or an artificial chromosome. Additional ways to confer accessory plasmid function to host cells will be evident to those of skill in the art, and the invention is not limited in this respect. 
     The term “mutagen,” as used herein, refers to an agent that induces mutations or increases the rate of mutation in a given biological system, for example, a host cell, to a level above the naturally-occurring level of mutation in that system. Some exemplary mutagens useful for continuous evolution procedures are provided elsewhere herein and other useful mutagens will be evident to those of skill in the art. Useful mutagens include, but are not limited to, ionizing radiation, ultraviolet radiation, base analogs, deaminating agents (e.g., nitrous acid), intercalating agents (e.g., ethidium bromide), alkylating agents (e.g., ethylnitrosourea), transposons, bromine, azide salts, psoralen, benzene, 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX) (CAS no. 77439-76-0), O,O-dimethyl-S-(phthalimidomethyl)phosphorodithioate (phos-met) (CAS no. 732-11-6), formaldehyde (CAS no. 50-00-0), 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (AF-2) (CAS no. 3688-53-7), glyoxal (CAS no. 107-22-2), 6-mercaptopurine (CAS no. 50-44-2), N-(trichloromethylthio)-4-cyclohexane-1,2-dicarboximide (captan) (CAS no. 133-06-2), 2-aminopurine (CAS no. 452-06-2), methyl methane sulfonate (MMS) (CAS No. 66-27-3), 4-nitroquinoline 1-oxide (4-NQO) (CAS No. 56-57-5), N4-aminocytidine (CAS no. 57294-74-3), sodium azide (CAS no. 26628-22-8), N-ethyl-N-nitrosourea (ENU) (CAS no. 759-73-9), N-methyl-N-nitrosourea (MNU) (CAS no. 820-60-0), 5-azacytidine (CAS no. 320-67-2), cumene hydroperoxide (CHP) (CAS no. 80-15-9), ethyl methanesulfonate (EMS) (CAS no. 62-50-0), N-ethyl-N-nitro-N-nitrosoguanidine (ENNG) (CAS no. 4245-77-6), N-methyl-N-nitro-N-nitrosoguanidine (MNNG) (CAS no. 70-25-7), 5-diazouracil (CAS no. 2435-76-9), and t-butyl hydroperoxide (BHP) (CAS no. 75-91-2). Additional mutagens can be used in continuous evolution procedures as provided herein, and the invention is not limited in this respect. 
     Ideally, a mutagen is used at a concentration or level of exposure that induces a desired mutation rate in a given host cell or viral vector population, but is not significantly toxic to the host cells used within the average time frame a host cell is exposed to the mutagen or the time a host cell is present in the host cell flow before being replaced by a fresh host cell. 
     The term “mutagenesis plasmid,” as used herein, refers to a plasmid comprising a gene encoding a gene product that acts as a mutagen. In some embodiments, the gene encodes a DNA polymerase lacking a proofreading capability. In some embodiments, the gene is a gene involved in the bacterial SOS stress response, for example, a UmuC, UmuD′, or RecA gene. In some embodiments, the gene is a GATC methylase gene, for example, a deoxyadenosine methylase (dam methylase) gene. In some embodiments, the gene is involved in binding of hemimethylated GATC sequences, for example, a seqA gene. In some embodiments, the gene is involved with repression of mutagenic nucleobase export, for example emrR. In some embodiments, the gene is involved with inhibition of uracil DNA-glycosylase, for example a Uracil Glycosylase Inhibitor (ugi) gene. In some embodiments, the gene is involved with deamination of cytidine (e.g., a cytidine deaminase from  Petromyzon marinus ), for example, cytidine deaminase 1 (CDA1). 
     The term “host cell,” as used herein, refers to a cell that can host a viral vector useful for a continuous evolution process as provided herein. A cell can host a viral vector if it supports expression of genes of viral vector, replication of the viral genome, and/or the generation of viral particles. One criterion to determine whether a cell is a suitable host cell for a given viral vector is to determine whether the cell can support the viral life cycle of a wild-type viral genome that the viral vector is derived from. For example, if the viral vector is a modified M13 phage genome, as provided in some embodiments described herein, then a suitable host cell would be any cell that can support the wild-type M13 phage life cycle. Suitable host cells for viral vectors useful in continuous evolution processes are well known to those of skill in the art, and the invention is not limited in this respect. 
     In some embodiments, modified viral vectors are used in continuous evolution processes as provided herein. In some embodiments, such modified viral vectors lack a gene required for the generation of infectious viral particles. In some such embodiments, a suitable host cell is a cell comprising the gene required for the generation of infectious viral particles, for example, under the control of a constitutive or a conditional promoter (e.g., in the form of an accessory plasmid, as described herein). In some embodiments, the viral vector used lacks a plurality of viral genes. In some such embodiments, a suitable host cell is a cell that comprises a helper construct providing the viral genes required for the generation of viral particles. A cell is not required to actually support the life cycle of a viral vector used in the methods provided herein. For example, a cell comprising a gene required for the generation of infectious viral particles under the control of a conditional promoter may not support the life cycle of a viral vector that does not comprise a gene of interest able to activate the promoter, but it is still a suitable host cell for such a viral vector. In some embodiments, the viral vector is a phage, and the host cell is a bacterial cell. In some embodiments, the host cell is an  E. coli  cell. Suitable  E. coli  host strains will be apparent to those of skill in the art, and include, but are not limited to, New England Biolabs (NEB) Turbo, Top10F′, DH12S, ER2738, ER2267, XL1-Blue MRF′, and DH10B. These strain names are art recognized, and the genotype of these strains has been well characterized. It should be understood that the above strains are exemplary only, and that the invention is not limited in this respect. 
     The term “fresh,” as used herein interchangeably with the terms “non-infected” or “uninfected” in the context of host cells, refers to a host cell that has not been infected by a viral vector comprising a gene of interest as used in a continuous evolution process provided herein. A fresh host cell can, however, have been infected by a viral vector unrelated to the vector to be evolved or by a vector of the same or a similar type but not carrying the gene of interest. In some embodiments, the host cell is a prokaryotic cell, for example, a bacterial cell, such as an  E. coli  cell. 
     In some embodiments, the host cell is an  E. coli  cell. In some embodiments of PACE, for example, in embodiments employing an M13 selection phage, the host cells are  E. coli  cells expressing the Fertility factor, also commonly referred to as the F factor, sex factor, or F-plasmid. The F-factor is a bacterial DNA sequence that allows a bacterium to produce a sex pilus necessary for conjugation and is essential for the infection of  E. coli  cells with certain phage, for example, with M13 phage. For example, in some embodiments, the host cells for M13-PACE are of the genotype F′proA + B + Δ(lacIZY) zzf::Tn10(TetR)/endA1 recA1 galE15 galK16 nupG rpsL ΔlacIZYA araD139 Δ(ara, leu)7697 mcrA Δ(mrr-hsdRMS-mcrBC) proBA::pir116 λ −− . In some embodiments, the host cells for M13-PACE are of the genotype F′proA+B+Δ(lacIZY) zzf::Tn10(TetR) lacIQ1PN25-tetR luxCDE/endA1 recA1 galE15 galK16 nupG rpsL(StrR) ΔlacIZYA araD139 Δ(ara,leu)7697 mcrA Δ(mrr-hsdRMS-mcrBC) proBA::pir116 araE201 ΔrpoZ Δflu ΔcsgABCDEFG ΔpgaC λ—, for example S1030 cells as described in Carlson, J. C., et al. Negative selection and stringency modulation in phage-assisted continuous evolution.  Nat. Chem. Biol.  10, 216-222 (2014). In some embodiments, the host cells for M13-PACE are of the genotype F′ proA+B+Δ(lacIZY) zzf::Tn10 lacIQ1 PN25-tetR luxCDE Ppsp(AR2) lacZ luxR Plux groESL/endA1 recA1 galE15 galK16 nupG rpsL ΔlacIZYA araD139 Δ(ara,leu)7697 mcrA Δ(mrr-hsdRMS-mcrBC) proBA::pir116 araE201 ΔrpoZ Δflu ΔcsgABCDEFG ΔpgaC λ—, for example 52060 cells as described in Hubbard, B. P. et al. Continuous directed evolution of DNA-binding proteins to improve TALEN specificity.  Nature Methods  12, 939-942 (2015). 
     The term “subject,” as used herein, refers to an individual organism, for example, an individual mammal. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human mammal. In some embodiments, the subject is a non-human primate. In some embodiments, the subject is a rodent. In some embodiments, the subject is a sheep, a goat, a cattle, a cat, or a dog. In some embodiments, the subject is a vertebrate, an amphibian, a reptile, a fish, an insect, a fly, or a nematode. In some embodiments, the subject is a research animal. In some embodiments, the subject is genetically engineered, e.g., a genetically engineered non-human subject. The subject may be of either sex and at any stage of development. In some embodiments, the subject has a disease characterized by increased activity of an intracellular protein (e.g., a SNARE protein, PTEN, etc.). In some embodiments, the disease characterized by increased activity of an intracellular protein is cancer, transplantation rejection, graft-versus-host disease, ischemic neuronal injury (stroke), Parkinson&#39;s Disease, Huntington&#39;s Disease, Alzheimer&#39;s Disease, spinal cord injury, diabetes, autoimmune disorders, allergy, or Cushing Disease. In some embodiments, the subject has a disease characterized by increased activity of a SNARE protein (e.g., VAMP1, VAMP2, VAMP7, VAMPS, SNAP23, SNAP25, etc.). In some embodiments, the subject has a disease characterized by increased activity of a VAMP protein (e.g., VAMP7). In some embodiments, the disease characterized by increased VAMP7 activity is cancer, transplantation rejection, or graft-versus-host disease. In some embodiments, the subject has a disease characterized by increased expression of a VAMP8 protein. In some embodiments, the disease characterized by increased VAMP8 activity is cancer. In some embodiments, the subject has or is suspected of having a disease characterized neuro-degeneration and in which inhibition of PTEN activity could transiently induce cellular regeneration, for example, ischemic neuronal injury (stroke), Parkinson&#39;s Disease, Huntington&#39;s Disease, Alzheimer&#39;s Disease, or spinal cord injury. In some embodiments, the subject has or is suspected of having a disease characterized by hypersecretion and in which cleavage of SNAP23 could prevent said secretion, for example, diabetes, autoimmune disorders, allergy, and Cushing Disease. 
     The term “cell,” as used herein, refers to a cell derived from an individual organism, for example, from a mammal. A cell may be a prokaryotic cell or a eukaryotic cell. In some embodiments, the cell is a eukaryotic cell, for example, a human cell, a mouse cell, a pig cell, a hamster cell, a monkey cell, etc. In some embodiments, a cell is characterized by increased VAMP7 expression, such as a neuronal cell. In some embodiments, a cell is obtained from a subject having or suspected of having a disease characterized by increased VAMP7 levels/expression, for example, cancer, transplantation rejection, or graft-versus-host disease, etc. In some embodiments, a cell is obtained from a subject having or suspected of having a disease characterized by increased PTEN levels/expression, for example, ischemic neuronal injury (stroke). 
     The term “intracellular environment,” as used herein refers, to the aqueous biological fluid (e.g., cytosol) forming the microenvironment contained by the outer membrane of a cell. For example, in a subject, an intracellular environment may include the cytoplasm of a cell or cells of a target organ or tissue (e.g., the cytosol of neuronal cells in CNS tissue). In another example, a cellular environment is the cytoplasm of a cell or cells surrounded by cell culture growth media housed in an in vitro culture vessel, such as a cell culture plate or flask. 
     The term “increased activity,” as used herein, refers to an increase in activity (e.g., via elevated expression) of a particular molecule in one cell or subject relative to a normal cell or subject that is not characterized by increased activity of that molecule (e.g., a “normal” or “control” cell or subject). For example, a cell having increased PTEN activity is characterized by more PTEN function (e.g., PTEN-mediated phosphatase activity) than a control cell having a normal (e.g., healthy) level of PTEN activity. In another example, a cell having increased VAMP7 activity is characterized by more VAMP7 activity (e.g., than a control cell expressing a normal (e.g., healthy) amount of VAMP7. Methods of determining relative expression levels of biomolecules (e.g., cytokines, proteins, nucleic acids, etc.) are known to the skilled artisan and include quantitative real-time PCR (q-RT-PCR), western blot, protein quantification assays (e.g., BCA assay), flow cytometry, etc. 
     As used herein, “aberrant activity” refers to an altered level of gene product (e.g., protein) activity in a cell or subject relative to a normal, healthy cell or subject. Examples of aberrant activity include but are not limited to increased activity of a gene product due to increased expression of the gene encoding the gene product, loss of activity of a gene product due to deceased expression of the gene encoding the gene product, altered function of a gene product due to epigenetic regulation of the gene encoding the gene product, etc., in a cell or subject relative to a normal, healthy cell or subject. 
     DETAILED DESCRIPTION 
     Introduction 
     Proteases are ubiquitous regulators of protein function in all domains of life and represent approximately one percent of known protein sequences. Substrate-specific proteases have proven useful as research tools and as therapeutics that supplement a natural protease deficiency to treat diseases, such as hemophilia, or that simply perform their native function such as in the case of botulinum toxin, which catalyzes the cleavage of SNARE proteins. 
     Researchers have engineered or evolved proteases for industrial use with enhanced thermostability and solvent tolerance. Similarly, a handful of therapeutic proteases have been engineered with improved kinetics and prolonged activity. The potential of proteases to serve as a broadly useful platform for degrading proteins implicated in disease, however, is greatly limited by the native substrate scope of known proteases. In contrast to the highly successful generation of therapeutic monoclonal antibodies with tailor-made binding specificities, the generation of proteases with novel protein cleavage specificities has proven to be a challenge. 
     The evolution of a protease that can degrade a target protein of interest often necessitates changing substrate sequence specificity at more than one position, and thus may require many generations of evolution. Continuous evolution strategies, which require little or no researcher intervention between generations, therefore may be well-suited to evolve proteases capable of cleaving a target protein that differs substantially in sequence from the preferred substrate of a wild-type protease. In phage-assisted continuous evolution (PACE), a population of evolving selection phage (SP) is continuously diluted in a fixed-volume vessel by an incoming culture of host cells, e.g.,  E. coli . The SP is a modified phage genome in which the evolving gene of interest has replaced gene III, a gene essential for phage infectivity. If the evolving gene of interest possesses the desired activity, it will trigger expression of gene III from an accessory plasmid (AP) in the host cell, thus producing infectious progeny encoding active variants of the evolving gene. The mutation rate of the SP is controlled using an inducible mutagenesis plasmid (MP), such as MP6 (for example as described in International PCT Application, PCT/US2016/027795, filed on Apr. 15, 2016, published as WO2016/168631 on Oct. 20, 2016, the entire contents of which are incorporated herein by reference), which upon induction increases the mutation rate of the SP by &gt;300,000-fold. Because the rate of continuous dilution is slower than phage replication but faster than  E. coli  replication, mutations only accumulate in the SP. 
     Some aspects of this disclosure are based on the recognition that PACE can be employed for the directed evolution of proteases, in particular the evolution of proteases that cleave intracellular proteins (e.g., VAMP1, VAMP2, VAMP7, VAMP8, SNAP23, PTEN, etc.). In some embodiments, proteases described by the disclosure are evolved from wild-type Botulinum toxin (BoNT) proteases, for example, BoNT E or BoNT F. Proteases may require many successive mutations to remodel complex networks of contacts with polypeptide substrates and are thus not readily manipulated by conventional, iterative evolution methods. The ability of PACE to perform the equivalent of hundreds of rounds of iterative evolution methods within days enables complex protease evolution experiments, that are impractical with conventional methods. 
     This disclosure provides data illustrating the feasibility of PACE-mediated evolution of the BoNT proteases (e.g., BoNT E or BoNT F) to cleave intracellular proteins (e.g., VAMP7, VAMP8, PTEN, etc.). As described in the Examples, BoNT E protease, which natively cleaves the consensus substrate sequence MGNEIDTQNRQIDRIMEKAD (SEQ ID NO: 310), was evolved by PACE to cleave a target sequence, NGSLCDQEIDSICSIERADN (SEQ ID NO: 311), that differs from the consensus substrate and is present in PTEN. Also described in the Examples, BoNT F protease, which natively cleaves the consensus substrate sequence TSNRRLQQTQAQVEEVVDIIRVNVDKVLERDQKLSELDDRADALQAGASQFESSAAKL KR (SEQ ID NO: 312), was evolved by PACE to cleave a target sequence, GGSGGSGGSKGLDKVMETQAQVDELKGIMVRNIDLVAQRGERLELLIDKTENLVDSSV TFKTTSRNLARGGSGGSGGS (SEQ ID NO: 313). It was observed that after constructing a pathway of evolutionary stepping-stones and performing ˜2,000 generations of evolution using PACE, the resulting BoNT protease variants (e.g., BoNT E variants and BoNT F variants) contain up to 20 amino acid substitutions relative to wild-type BoNT proteases (e.g., SEQ ID NO: 286 or 287) and cleave human VAMP7 or PTEN at the intended target peptide bond. Together, the work described herein establishes a platform for generating proteases (e.g., BoNT protease variants) with changed substrate specificities and the ability to cleave proteins implicated in human disease. 
     PACE technology has been described previously, for example, in International PCT Application, PCT/US2009/056194, filed Sep. 8, 2009, published as WO 2010/028347 on Mar. 11, 2010; International PCT Application, PCT/US2011/066747, filed Dec. 22, 2011, published as WO 2012/088381 on Jun. 28, 2012; and U.S. Application, U.S. Ser. No. 13/922,812, filed Jun. 20, 2013; International PCT Application, PCT/US2015/057012, filed on Oct. 22, 2015, published as WO2016/077052; and, PCT/US2016/027795, filed on Apr. 15, 2016, published as WO2016/168631, each of which is incorporated herein by reference. Those of skill in the art will understand that the PACE technology, strategies, methods, compositions, systems, and reagents provided herein can be used in combination with many aspects of the PACE technology described in those applications, for example, with the apparatuses, lagoons, host cell types, cell flow parameters, selection stringencies (e.g., high selection stringency, low selection stringency, etc.), positive selection strategies, negative selection strategies, plasmids, vectors, etc., disclosed in those applications. 
     Variant BoNT Proteases and Uses Thereof 
     This disclosure provides variants of BoNT proteases that are derived from a wild-type BoNT E or BoNT F protease (e.g., SEQ ID NO: 286 or 287) and have at least one of the amino acid variations present in Table 1 or Table 2. The variation in amino acid sequence generally results from a mutation, insertion, or deletion in a DNA coding sequence. Mutation of a DNA sequence can result in a nonsense mutation (e.g., a transcription termination codon (TAA, TAG, or TAA) that produces a truncated protein), a mis sense mutation (e.g., an insertion or deletion mutation that shifts the reading frame of the coding sequence), or a silent mutation (e.g., a change in the coding sequence that results in a codon that codes for the same amino acid normally present in the cognate protein, also referred to sometimes as a synonymous mutation). In some embodiments, mutation of a DNA sequence results in a non-synonymous (i.e., conservative, semi-conservative, or radical) amino acid substitution. 
     Generally, wild-type BoNT protease is encoded by a gene of the microorganism  Clostridium botulinum . The amount or level of variation between a wild-type BoNT protease and a BoNT protease variant provided herein can be expressed as the percent identity of the nucleic acid sequences or amino acid sequences between the two genes or proteins. In some embodiments, the amount of variation is expressed as the percent identity at the amino acid sequence level. In some embodiments, a BoNT protease variant and a wild-type BoNT protease are from about 70% to about 99.9% identical, about 75% to about 95% identical, about 80% to about 90% identical, about 85% to about 95% identical, or about 95% to about 99% identical at the amino acid sequence level. In some embodiments, a BoNT protease variant comprises an amino acid sequence that is at least 95%, at least 99%, or at least 99.9% identical to the amino acid sequence of a wild-type BoNT protease. 
     In some embodiments, a variant BoNT protease is about 70%, about 71%, about 72%, about 73%, about 74%, about 75%, about 76%, about 77%, about 78%, about 79%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 99.9% identical to a wild-type BoNT protease. 
     Some aspects of the disclosure provide variant BoNT proteases having between about 90% and about 99.9% (e.g., about 90%, about 90.5%, about 91%, about 91.5%, about 92%, about 92.5%, about 93%, about 93.5%, about 94%, about 94.5%, about 95%, about 95.5%, about 96%, about 96.5%, about 97%, about 97.5%, about 98%, about 98.5%, about 99%, about 99.2%, about 99.4%, about 99.6%, about 99.8%, or about 99.9%) identical to a wild-type BoNT protease as set forth in SEQ ID NO: 286 or 287. In some embodiments, the variant BoNT protease is no more than 99.9% identical to a wild-type BoNT protease. 
     Some aspects of the disclosure provide variant BoNT proteases having between 1 and 20 amino acid substitutions (e.g., mutations) relative to a wild-type BoNT protease (e.g., SEQ ID NO: 286 or 287). In some embodiments, a variant BoNT protease has 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acid substitutions relative to a wild-type BoNT protease (e.g., SEQ ID NO: 286 or 287). 
     The amount or level of variation between a wild-type BoNT protease and a variant BoNT protease can also be expressed as the number of mutations present in the amino acid sequence encoding the variant BoNT protease relative to the amino acid sequence encoding the wild-type BoNT protease. In some embodiments, an amino acid sequence encoding a variant BoNT protease comprises between about 1 mutation and about 100 mutations, about 10 mutations and about 90 mutations, about 20 mutations and about 80 mutations, about 30 mutations and about 70 mutations, or about 40 and about 60 mutations relative to an amino acid sequence encoding a wild-type BoNT protease. In some embodiments, an amino acid sequence encoding a variant BoNT protease comprises more than 100 mutations relative to an amino acid sequence encoding a wild-type BoNT protease. In some embodiments, the variant BoNT protease comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35 amino acid variations selected from the variations (e.g., amino acid substitutions) provided in Table 1. In some embodiments, the variant BoNT protease comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35 amino acid variations selected from the variations (e.g., amino acid substitutions) provided in Table 2. 
     
       
         
           
               
             
               
                 TABLE 1 
               
               
                   
               
               
                 Unique BoNT E Mutations 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 Q354R 
                 T400K 
                 Y355H 
                 Y357S 
               
               
                   
                 G101S 
                 V265G 
                 F186L 
                 C26Y 
               
               
                   
                 D312N 
                 I409T 
                 N118D 
                 N238S 
               
               
                   
                 V345I 
                 T400A 
                 Q27R 
                 S198G 
               
               
                   
                 M172R 
                 I247V 
                 Q27H 
                 Y20C 
               
               
                   
                 M172S 
                 T160S 
                 A224S 
                 I21M 
               
               
                   
                 M172V 
                 K225Y 
                 N138D 
                 L98P 
               
               
                   
                 I227T 
                 K245R 
                 N161W 
                 E28K 
               
               
                   
                 I227V 
                 Q141R 
                 N161V 
                 I409L 
               
               
                   
                 S372G 
                 S99T 
                 Y68H 
                 Y357C 
               
               
                   
                 A266T 
                 A313S 
                 Y171C 
                 I352T 
               
               
                   
                 I263V 
                 V132G 
                 S6G 
                 A129Q 
               
               
                   
                 F358L 
                 C231G 
                 E159S 
                 S162A 
               
               
                   
                 N197K 
                 N379K 
                 E159R 
                 Y357H 
               
               
                   
                 M172K 
                 D128G 
                 I352V 
                 S174A 
               
               
                   
                 Q295R 
                 K311E 
                 E159W 
                 E159Q 
               
               
                   
                 S99A 
                 S314A 
                 I232T 
                 Y357W 
               
               
                   
                 N161Y 
                 K225H 
                 V47I 
                 L404* 
               
               
                   
                 E154G 
                 C231R 
                 E78G 
                 E148G 
               
               
                   
                 E159L 
                 Q27K 
                 S137R 
                 Y357P 
               
               
                   
                 N242S 
                 K225L 
                 E159A 
                 I302M 
               
               
                   
                 D128A 
                 K22S 
                 E159C 
                 I399S 
               
               
                   
                 I199M 
                 K22R 
                 I352A 
                 E184G 
               
               
                   
                 T160A 
                 K329N 
                 S163R 
                 T119K 
               
               
                   
                 A389T 
                 D53Y 
                 R168K 
                 S162Q 
               
               
                   
                 I165V 
                 N161H 
                 I232S 
                 A129G 
               
               
                   
                 I18V 
                 E89P 
                 Q141K 
                 S187F 
               
               
                   
                 I199T 
                 K225E 
                 I233T 
                 N258D 
               
               
                   
                 N261D 
                 D270N 
                 I396S 
                 P398L 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 2 
               
               
                   
               
               
                 Unique BoNT F Mutations 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 A63D 
                 I139V 
                 V422I 
                 N6S 
               
               
                   
                 E105A 
                 N165T 
                 V422L 
                 Y10C 
               
               
                   
                 Y113H 
                 N165S 
                 K283E 
                 R49L 
               
               
                   
                 Y113D 
                 N358T 
                 F267L 
                 I52V 
               
               
                   
                 S70F 
                 D175A 
                 N305T 
                 D58Y 
               
               
                   
                 A63T 
                 D161N 
                 S136I 
                 E60D 
               
               
                   
                 E66D 
                 P221R 
                 D382Y 
                 E66K 
               
               
                   
                 T79S 
                 N101T 
                 S176G 
                 S70H 
               
               
                   
                 K96N 
                 Y113S 
                 A291V 
                 T90I 
               
               
                   
                 K172R 
                 I277L 
                 L421W 
                 T123M 
               
               
                   
                 N76D 
                 S167C 
                 K146R 
                 T123S 
               
               
                   
                 R303H 
                 N101I 
                 S176N 
                 T132I 
               
               
                   
                 S189I 
                 S30N 
                 Y316N 
                 V145I 
               
               
                   
                 S244I 
                 K342R 
                 E310G 
                 G159S 
               
               
                   
                 H129Y 
                 D60Y 
                 G241S 
                 D161G 
               
               
                   
                 Y210C 
                 D185A 
                 Y294C 
                 N184K 
               
               
                   
                 V426* 
                 A330E 
                 D55N 
                 Y201H 
               
               
                   
                 N184H 
                 I385V 
                 K411E 
                 N211S 
               
               
                   
                 E338K 
                 I39V 
                 A307S 
                 F217L 
               
               
                   
                 Y294H 
                 R244C 
                 S100I 
                 A226S 
               
               
                   
                 G420A 
                 S213Y 
                 F341L 
                 A232T 
               
               
                   
                 G420C 
                 D60E 
                 1416V 
                 I262T 
               
               
                   
                 Y199H 
                 E164G 
                 A292S 
                 L264M 
               
               
                   
                 G420V 
                 N11S 
                 K411N 
                 D274M 
               
               
                   
                 A82V 
                 E259G 
                 G209D 
                 N314S 
               
               
                   
                 Q109H 
                 Y244C 
                 Y253S 
                 G325S 
               
               
                   
                 P197S 
                 E164K 
                 D175G 
                 D331G 
               
               
                   
                 S350I 
                 T243A 
                 S415P 
                 S333F 
               
               
                   
                 E200K 
                 T214A 
                 D274E 
                 T335I 
               
               
                   
                 N339T 
                 D355G 
                 E215K 
                 N339S 
               
               
                   
                 N379D 
                 T279C 
                 I370V 
                 F360I 
               
               
                   
                 R41H 
                 F374L 
                 S207N 
                 T367S 
               
               
                   
                 R300H 
                 V362A 
                 I425T 
                 F369F 
               
               
                   
                 E200A 
                 K371E 
                 I425S 
                 V377I 
               
               
                   
                 E121K 
                 N329D 
                 N211D 
                 N379H 
               
               
                   
                 E423R 
                 Y201D 
                 A103V 
                 K29E 
               
               
                   
                 E121D 
                 T214G 
                 K347N 
                 K31N 
               
               
                   
                 N184S 
                 N40T 
                 S389N 
                 Y72H 
               
               
                   
                 I190V 
                 T214S 
                 I286N 
                 N99S 
               
               
                   
                 F428S 
                 L375R 
                 D414G 
                 V106A 
               
               
                   
                 Y237S 
                 N211H 
                 A63V 
                 Y113C 
               
               
                   
                 R303C 
                 T299M 
                 M174T 
                 V131G 
               
               
                   
                 T335S 
                 R240L 
                 G177A 
                 S141T 
               
               
                   
                 S350G 
                 R240F 
                 G178A 
                 I150T 
               
               
                   
                 F360L 
                 F267I 
                 N184T 
                 V155I 
               
               
                   
                 Y372H 
                 F270V 
                 V193M 
                 S166Y 
               
               
                   
                 N396H 
                 N293D 
                 E200G 
                 S167I 
               
               
                   
                 P410L 
                 I297L 
                 Y210H 
                 T214I 
               
               
                   
                 D418Y 
                 E215G 
                 E423K 
                 N409D 
               
               
                   
                 F420S 
                 S224I 
                 E423K 
                 S224V 
               
               
                   
                   
               
            
           
         
       
     
     Particular combinations of mutations present in an amino acid sequence encoding a variant BoNT protease can be referred to as the “genotype” of the variant BoNT protease. For example, a variant BoNT E protease genotype may comprise the mutations Q27H, S99A, G101S, N118D, E159L, N161Y, S162Q, S163R, M172K, I132T, Q354R, Y357P, relative to a wild-type BoNT E protease (e.g., SEQ ID NO: 286; wild-type BoNT E). In some embodiments, a BoNT F variant genotype comprises the following mutations: V106A, S166Y, S1671, E200G, S224I R240L, S350G, F360L, Y372H, P410L, G420A, L421W, V422L, E423R, I425S, and V426* (e.g., a stop codon). In some embodiments, a BoNT F variant genotype comprises the following mutations: S166Y, N184K, E200G, S224I, R240F, T335S, F360L Y372H, N396H, P410L, D418Y, and E423K. Further examples of variant BoNT protease genotypes are shown in Tables 8-27, 29 and 30. 
     In some embodiments, a variant BoNT protease comprises at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 mutations provided in Table 1 or Table 2. In some embodiments, the at least one mutation is selected from the group consisting of: 118V, C26Y, Q27H, E28K, F29L, Y68H, L89P, S99A, S99T, G101S, N118D, G127S, Q141K, E154G, E159L, N161Y, S162Q, S163R, R168K, M172K, K225E, C231R, I232T, I233T, N238S, Q295R, I396S, P398L Q354R, Y357P, L404*, and I409T, and I409T. In some embodiments, the at least one mutation is selected from the group consisting of: Q27H, S99A, G101S, N118D, E159L, N161Y, S162Q, S163R, M172K, I132T, Q354R, and Y357P. In some embodiments, a variant BoNT protease as described herein comprises or consists of a sequence selected from SEQ ID NOs: 1-285 given in Table 28. In some embodiments, the lowercase amino acid residues indicate the amino acid substitutions. 
     
       
         
           
               
               
             
               
                 TABLE 28 
               
               
                   
               
               
                 Sequence 
                 SEQ ID NO 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
            
               
                 MPKINTSFNYNDPVNDRTILYIKPGGCkEFYKSFNIMKNIWIIPERNVIGTT 
                 1 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCsITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINTSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 2 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYtGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINTSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 3 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCkEFYKSFNIMKNIWIIPERNVIGTT 
                 4 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 5 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 6 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 7 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFaThSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCsITQQQNPLITNRKGINIEEFLTF 
                   
               
               
                 GGNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQ 
                   
               
               
                 EKYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCkEFYKSFNIMKNIWIIPERNVIGTT 
                 8 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFaThSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCkEFYKSFNIMKNIWIIPERNVIGTT 
                 9 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCsITQQQNPLITNRKGINIEEFLTF 
                   
               
               
                 GGNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQ 
                   
               
               
                 EKYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 10 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFaTNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 11 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFlTySSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQY 
                   
               
               
                 KYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 12 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFsThSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 13 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFaTySSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILcIKPGGCQkFYKSFNIMKNIWIIPERNVIGTTP 
                 14 
               
               
                 QDFHPPTSLKNGDSSYYDPNYLQSDgEKDRFLKIVTKIFNRINNNLSGGI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGaqSAVEIKFSNGSQHILLPNVIIMGA 
                   
               
               
                 EPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQiKCRETYIGQYK 
                   
               
               
                 YFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRG*VKK 
                   
               
               
                 IIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 15 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGggSAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPsTGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 16 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGgqSAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 17 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGaqSAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGtTTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 18 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGaqSAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPIaGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 19 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 20 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 21 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNvSLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 22 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGsI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLkNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 23 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNpSGGI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYlKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 24 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCsITQQQNPLITNRKGINIEEFLTF 
                   
               
               
                 GGNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQ 
                   
               
               
                 EKYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPIkGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 25 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 26 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGsI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLkNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 27 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCrEFYKSFNIMKNIWIIPERNVIGTTP 
                 28 
               
               
                 QDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNpSGGI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDkSmNEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 29 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGvTTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYtGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRG*V 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 30 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNcMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGshGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIsPGGCQkFYKSFNIMKNIWIIPERNVIGTTP 
                 31 
               
               
                 QDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGGI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIgTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKcFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 32 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 33 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrh 
                   
               
               
                 KYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 34 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITgAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQY 
                   
               
               
                 KYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNiIGTTP 
                 35 
               
               
                 QDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNpSGGI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNIgEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 36 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrh 
                   
               
               
                 KYFKLSNLLNDSIYNISEGYNINkLKVNFRGQNANLNPRIIKPITGRGLVK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 37 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDkSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDAaGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrh 
                   
               
               
                 KYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 38 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFsThSSNISLRNNYsPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAhGITTTgIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQY 
                   
               
               
                 KYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 39 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFsTwSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYtGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIlRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCrEFYKSFNIMKNIWIIPERNVIGTTP 
                 40 
               
               
                 QDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNpSGGI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFwTwSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 41 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFqTySSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 42 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFaTwSSNISLRNNYvPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAlGITTTrIITQQQNPLITNRKGINIEEFLTFGGN 
                   
               
               
                 DLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEKY 
                   
               
               
                 GLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQYK 
                   
               
               
                 YFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCrEFYKSFNIMKNIWIIPERNVIGTTP 
                 43 
               
               
                 QDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNpSGGI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFaTvSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 44 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNpSGGI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFcTwSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTrIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQY 
                   
               
               
                 KYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 45 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 46 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 47 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFaThSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAyGITTTCtITQQQNPLIsNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 48 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNgINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 49 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFaThSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 50 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNgIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKhFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 51 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNgIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKcFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 52 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNgIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTlSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKcFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 53 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNgIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNcMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 54 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSrHILLPNVIIMG 
                   
               
               
                 AEPDLFaahSSNISLRNNYMPaNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrhK 
                   
               
               
                 YFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 55 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFaahSSNISLRNNYMPaNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrhK 
                   
               
               
                 YFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRG*VKK 
                   
               
               
                 IIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 56 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFaahSSNISLRNNYMPaNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrhK 
                   
               
               
                 YFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 57 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFaahSSNISLRNNYMPaNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKnASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrhKY 
                   
               
               
                 FKLSNLLNDSIYNISEGYNINNLKVNFRGQNtNLNPRIIKPITGRGLVKKII 
                   
               
               
                 RF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 58 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFcswSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTrIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQY 
                   
               
               
                 KsFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 59 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFcTwSSNISLRNNYrPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAlGITTTrIITQQQNPLITNRKGINIEEFLTFGGN 
                   
               
               
                 DLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEKY 
                   
               
               
                 GLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQYKc 
                   
               
               
                 FKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVKKI 
                   
               
               
                 IRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 60 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFrTySSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDkSmNEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDeDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQY 
                   
               
               
                 KYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 61 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFcswSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTrIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQY 
                   
               
               
                 KcFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 62 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFcTwSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTrIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQY 
                   
               
               
                 KcFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 63 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFcswSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTrIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQY 
                   
               
               
                 KYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 64 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFsTwSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDkSmNEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 65 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 66 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVtQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQY 
                   
               
               
                 KYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 67 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSdGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILnKLYSFTEFDLATKFQVKCRETYIGQY 
                   
               
               
                 KYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 68 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFrNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 69 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AgPDLFEaNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTrIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIvTVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKcFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIrPGGCQkFYKSFNIMKNIWIIPERNVIGTTP 
                 70 
               
               
                 QDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGGI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AgPDLFETNSrNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYaGQ 
                   
               
               
                 YKcFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIrPGGCQkFYKSFNIMKNIWIIPERNVIGTTP 
                 71 
               
               
                 QDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGGI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AgPDLFETNSrNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYaGQ 
                   
               
               
                 YKwFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGL 
                   
               
               
                 VKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 72 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AgPDLFEaNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAhGITTTrIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIvTVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQ 
                   
               
               
                 YKcFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPsTGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 73 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AgPDLFETNaSNISLRNNYrPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAhGITTTCIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQY 
                   
               
               
                 KYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 74 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFaahSSNISLRNNYMPaNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrhK 
                   
               
               
                 YFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRG*VKK 
                   
               
               
                 IIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 75 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGgASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFaahSSNISLRNNYMPaNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrhK 
                   
               
               
                 YFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRG*VKK 
                   
               
               
                 IIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYmKPGGCQkFYKSFNIMKNIWIIPERNVIGT 
                 76 
               
               
                 TPQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSG 
                   
               
               
                 GILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIM 
                   
               
               
                 GAEPDLFaahSSNISLRNNYMPaNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrh 
                   
               
               
                 KYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRG*VK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 77 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFaahSrNISLRNNYMPaNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRrGINIEEFLTFGG 
                   
               
               
                 NDLNIITgAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEKY 
                   
               
               
                 GLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrhKY 
                   
               
               
                 FKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVKKI 
                   
               
               
                 IRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGChEFYKSFNIMKNIWIIPERNVIGTT 
                 78 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLaGsI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFlTyqSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrhK 
                   
               
               
                 YFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 79 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNkPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFaahSSNISLRNNYMPaNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrhK 
                   
               
               
                 YFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRG*VKK 
                   
               
               
                 IIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 80 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFcTwSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTrIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrhK 
                   
               
               
                 YFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 81 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFcswSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSmNEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTrIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITgAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEKY 
                   
               
               
                 GLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrYKcF 
                   
               
               
                 KLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVKKII 
                   
               
               
                 RF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 82 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFcswSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTrIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQY 
                   
               
               
                 KcFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 83 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFcTwSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGslGITTTrIITQQQNPLITNRrGINIEEFLTFGGN 
                   
               
               
                 DLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEKY 
                   
               
               
                 GLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrhKY 
                   
               
               
                 FKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVKKI 
                   
               
               
                 IRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 84 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFcswSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSmNEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTrIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITgAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEKY 
                   
               
               
                 GLDKDsSGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrYKcF 
                   
               
               
                 KLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVKKII 
                   
               
               
                 RF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 85 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFcswSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTrIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQY 
                   
               
               
                 KcFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRG*VK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 86 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFcTwSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTrIITQQQNPLITNRKGvNIEEFLTFG 
                   
               
               
                 GNDLNIITgAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYvGQY 
                   
               
               
                 KcFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTT 
                 87 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFcswSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSmNEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTrIITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITgAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEKY 
                   
               
               
                 GLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGQYKc 
                   
               
               
                 FKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVKKI 
                   
               
               
                 IRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 88 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrh 
                   
               
               
                 KYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINgFNYNDPVNDRTILYIKPGGCQkFYKSFNIMKNIWIIPERNVIGTT 
                 89 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGG 
                   
               
               
                 ILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQ 
                   
               
               
                 DPALTLMHELIHSLHGLYGAlGITTTCtITQQQNPLITNRKGINIEEFLTFG 
                   
               
               
                 GNDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQE 
                   
               
               
                 KYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrh 
                   
               
               
                 KYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLV 
                   
               
               
                 KKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGChEFYKSFNIMKNIWIIPERNVIGTT 
                 90 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLaGsI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFlTyqSNISLRNNYkPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDmFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrhK 
                   
               
               
                 YFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGChEFYKSFNIMKNIWIIPERNVIGTT 
                 91 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLaGsI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFlTyqSNISLRNNYkPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrhK 
                   
               
               
                 YFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGChEFYKSFNIMKNIWIIPERNVIGTT 
                 92 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLaGsI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAgEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFlTyqSNISLRNNYkPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrhK 
                   
               
               
                 YFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGChEFYKSFNIMKNIWIIPERNVIGTT 
                 93 
               
               
                 PyDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLaGsI 
                   
               
               
                 LLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFlTyqSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQD 
                   
               
               
                 PALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrhK 
                   
               
               
                 YFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVK 
                   
               
               
                 KIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGChEFYKSFNIMKNIWIIPERNVIGTT 
                 94 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLaGsI 
                   
               
               
                 LLEELSKANPYLGNDdTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFlTyqrNISLRNNYkPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQDP 
                   
               
               
                 ALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrYK 
                   
               
               
                 pFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVKK 
                   
               
               
                 IIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGyhEFYKSFNIMKNIWIIPERNVIGTTP 
                 95 
               
               
                 QDFHPPTSLKNGDSShYDPNYLQSDEEKDRFLKIVTKIFNRINNNLaGsIL 
                   
               
               
                 LEELSKANPYLGNDdTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMGA 
                   
               
               
                 EPDLFlTyqrNISLRNNYkPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQDPA 
                   
               
               
                 LTLMHELIHSLHGLYGAKGITTTCtITQQQsPLITNRKGINIEEFLTFGGND 
                   
               
               
                 LNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEKYGL 
                   
               
               
                 DKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrYKhFK 
                   
               
               
                 LSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVKKIIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGChEFYKSFNIMKNIWIIPERNVIGTT 
                 96 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLaGsI 
                   
               
               
                 LLEELSKANPYLGNDdTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFlTyqrNISLRNNYkPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQDP 
                   
               
               
                 ALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrYK 
                   
               
               
                 pFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVKK 
                   
               
               
                 ItRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGChEFYKSFNIMKNIWIIPERNVIGTT 
                 97 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLtGsI 
                   
               
               
                 LLEELSKANPYLGNDdTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFlTyqrNISLRNNYkPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQDP 
                   
               
               
                 ALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrYK 
                   
               
               
                 pFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRG*VKK 
                   
               
               
                 IIRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGyhEFYKSFNIMKNIWIIPERNVIGTTP 
                 98 
               
               
                 QDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLaGsIL 
                   
               
               
                 LEELSKANPYLGNDdTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMGA 
                   
               
               
                 EPDLFlTyqrNISLRNNYkPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQDPA 
                   
               
               
                 LTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFGGN 
                   
               
               
                 DLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEKY 
                   
               
               
                 GLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrYKh 
                   
               
               
                 FKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVKKI 
                   
               
               
                 IRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGChEFYKSFNIMKNIWIIPERNVIGTT 
                 99 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLaGsI 
                   
               
               
                 LLEELSKANPYLGNDdTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFlTyqrNISLRNNYkPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQDP 
                   
               
               
                 ALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPrLNPYKDIFQEKY 
                   
               
               
                 GLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrYKp 
                   
               
               
                 FKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVKKI 
                   
               
               
                 IRF 
                   
               
               
                   
               
               
                 MPKINSFNYNDPVNDRTILYIKPGGChEFYKSFNIMKNIWIIPERNVIGTT 
                 100 
               
               
                 PQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLaGsI 
                   
               
               
                 LLEELSKANPYLGNDdTPDNQFHIGDASAVEIKFSNGSQHILLPNVIIMG 
                   
               
               
                 AEPDLFlTyqrNISLRNNYkPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQDP 
                   
               
               
                 ALTLMHELIHSLHGLYGAKGITTTCtITQQQNPLITNRKGINIEEFLTFGG 
                   
               
               
                 NDLNIITVAQYNDIYTNLLNDYRKIASKLSKVQVSNPQLNPYKDIFQEK 
                   
               
               
                 YGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVKCRETYIGrYK 
                   
               
               
                 hFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVKK 
                   
               
               
                 IIRF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 101 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlnIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 102 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 103 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 104 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVrSSIIL 
                   
               
               
                 NLLVLGAGPDIFEsySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 105 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVrSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 106 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 107 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPdLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 108 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPdLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 109 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 110 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSsDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 111 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 112 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMaSGGVYDPSkDGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 113 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 114 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkaGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 115 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDvEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKvLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 116 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDvEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 117 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFgNySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMhNVWIIPER 
                 118 
               
               
                 NTIGTDPSDFePPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNEtKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 119 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDvYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 120 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLiLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNleIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 121 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEIShAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFgNySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 122 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 123 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFEsySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 124 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 125 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFEsySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKt*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKnKKYYKAFEIMRNVWIIPER 
                 126 
               
               
                 NTIGTDPSDFDPPASLdNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSaESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 127 
               
               
                 NTIGTDPSDFDPPASLdNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMgSGGVYDPSkDGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 128 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 129 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 130 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENyiYPVRKLMDSGGVYDPSkDGFGiINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 131 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENyiYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 132 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENyiYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKlNEIYKKLYgF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 133 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 134 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkaGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANhEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 135 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDvEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMhNVWIIPER 
                 136 
               
               
                 NTIGTDPSDFePPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEnKDYFQWKYGLDKNADGSYTVNENKFrEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEvMRNVWIIPER 
                 137 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDvEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 138 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFvKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 139 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFEtytYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYTF 
                   
               
               
                 NDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNRGQ 
                   
               
               
                 NIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 140 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 141 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFEsySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYsVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLAtKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNRG 
                   
               
               
                 QNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 142 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENytYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNRG 
                   
               
               
                 QNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 143 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENytYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYsVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNRG 
                   
               
               
                 QNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 144 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENytYPVRrLMDSGGVYDPSkDGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNRG 
                   
               
               
                 QNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 145 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENytYPVRKLMDSGGVYDPSkDGFGSINIVTFSsEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNRG 
                   
               
               
                 QNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 146 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLsTDAEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENytYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNRG 
                   
               
               
                 QNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 147 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENytYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLsNYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNRG 
                   
               
               
                 QNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 148 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFENyiYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 149 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 150 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTiVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLyDDIYTVSEGFNIGhLAVNNRG 
                   
               
               
                 QNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 151 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMeEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNEtKFNEIYKKLYSFTE 
                   
               
               
                 IDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNRG 
                   
               
               
                 QNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 152 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLvANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 153 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKnLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 154 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSh 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 155 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNdHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFEtytYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYTF 
                   
               
               
                 NDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNRGQ 
                   
               
               
                 NIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 156 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 157 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFEsyiYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYTF 
                   
               
               
                 NDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNRGQ 
                   
               
               
                 NIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 158 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFEtytYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYTF 
                   
               
               
                 NDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNRGQ 
                   
               
               
                 NIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 159 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFEtytYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYTF 
                   
               
               
                 NDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNRGQ 
                   
               
               
                 NIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 160 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFEtytYPVRrLMDSGGVYDPSkDGFGSINIVTFSPEYEYTF 
                   
               
               
                 NDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVnEGFNIGhLAVNNRGQ 
                   
               
               
                 NIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 161 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNkHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFEtytYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYTF 
                   
               
               
                 NDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNRGQ 
                   
               
               
                 NIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 162 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNkHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFEtytYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEYTF 
                   
               
               
                 NDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQeLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNRGQ 
                   
               
               
                 NIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 163 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNkHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFEtytYPVRKLMDSGGVYDPSkDGFGSINImTFSPEYEYTF 
                   
               
               
                 NDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQsP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNRGQ 
                   
               
               
                 NIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 164 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 165 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYsVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 166 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 167 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGcNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 168 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKnLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 169 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIIcSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 170 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPnIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIAmRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 171 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSsESFIADPAISLAHELIHALHGLYGARGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDpIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 172 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLsGARGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKFKVKCRNTYFIKYGFLKVPdLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 173 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIAmRL 
                   
               
               
                 SRVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLY 
                   
               
               
                 SFTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAV 
                   
               
               
                 NNRGQNIKLNPKIIDSvPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 174 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 175 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEd 
                   
               
               
                 TFNDISGGYdSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNeDGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKFKVKCRNTYFIKhGFLKVPdLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 176 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSsESFIADPAISLAHELIHALHGLYGARGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 \ASAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 177 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDnGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKdADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPdLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMhNVWIIPER 
                 178 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPdLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMhNVWIIPER 
                 179 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNkNKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPdLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 180 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPdLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 181 
               
               
                 NTIGTnPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFnRINS 
                   
               
               
                 NPAGEVLLQEISsAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIILN 
                   
               
               
                 LLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEElLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIgSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 182 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPShDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKnYNNLLANYEKIATRL 
                   
               
               
                 SRVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLY 
                   
               
               
                 SFTEIDLANKFKVKCRNTYFIKYGFLKVPdLLDDDIYTVSEGFNIGNLAV 
                   
               
               
                 NNRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 183 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEElLTFGGQDLNIITSAMKEKIYNNLLANYEKIAThLSh 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKFKVKCRNTYFIKYGlLKVPdLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMhNVWIIPER 
                 184 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNPKIIgSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 185 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKFKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 186 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMhNVWIIPER 
                 187 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAgKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNPKIIgSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 188 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAgKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMhNVWIIPER 
                 189 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLhEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKaKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNPKIIgSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 190 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINi 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 191 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNdHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEElLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIgLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 192 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 193 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPvGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 194 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNeDGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 195 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKs 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 196 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADrAISLAHELIHALHGLYGARGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 197 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSsPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 198 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLsNYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 199 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINi 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 200 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNdHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEElLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIgLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 201 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 202 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPvGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 203 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNeDGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 204 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKs 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 205 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADrAISLAHELIHALHGLYGARGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 206 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSsPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 207 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLsNYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 208 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENSSYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 209 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 210 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 StPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 211 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINT 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENSSYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 212 
               
               
                 NTIGTDPSDFDPPtSLdNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMgSGGVYDPSkDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 213 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINT 
                   
               
               
                 SNPAGEaLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYkY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKvLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 214 
               
               
                 NTIGTDPSDFDPPASLkNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPShDGFGSINIVTFSPEYkY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVaYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKvLiEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 215 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINT 
                   
               
               
                 SNPAGEaLLQEISsAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDgGGVYDPSNDGFGSINIVTFSPEYkY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGwlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 216 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINT 
                   
               
               
                 SNPAGEaLLQEISsAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPShDGFGSINIVTFSPEYkY 
                   
               
               
                 TFNDISGdYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGwlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYsDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPERN 
                 217 
               
               
                 TIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEaLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIILN 
                   
               
               
                 LLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSaESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKvLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 218 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINT 
                   
               
               
                 SNPAGEaLLQEISdAKPYLGNEHTPINEFyPVTRTTSVNIKSSTNVKSSIILN 
                   
               
               
                 LLVLGAGPDIFENySYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKvLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 219 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINT 
                   
               
               
                 SNPAGEVLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPShDGFGSINIVTFSPEYkY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 220 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINT 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSsDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 221 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINT 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSsDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 222 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINT 
                   
               
               
                 SNPAGEVLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENycYPVRKLMDSGGVYDPShDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 223 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPShDGFGSINIVTFSPEYkY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNlITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 224 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPdYLTTDAEKDRYLKTTIKLFKRINTS 
                   
               
               
                 NPAGEaLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIILN 
                   
               
               
                 LLVLGAGPDIFENySYPVRKLMDSGGVYDPShDGFGSINIVTFSPEYkYTF 
                   
               
               
                 NDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNRG 
                   
               
               
                 QNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 225 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTcKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYiF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 226 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTcKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 227 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTcKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 228 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENSSYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 229 
               
               
                 NTIGTDPSDFDPPASLdNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMgSGGVYDPSkDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAIiLAHELIHALHGLYGAfGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 230 
               
               
                 NTIGTDPSDFDPPASLdNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMgSGGVYDPSkDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAIiLAHELIHALHGLYGAfGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPdLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 231 
               
               
                 NTIGTDPSDFDPPASLdNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMgSGGVYDPSkDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSgESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIehGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 232 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENyiYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYhSSTESFIADPAIiLAHELIHALHGLYGAlGVTYKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNRGQ 
                   
               
               
                 NIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 233 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENyiYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAIiLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 234 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPdYLTTDAEKDRYLKTTIKLFKRINTS 
                   
               
               
                 NPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIILN 
                   
               
               
                 LLVLGAGPDIFENyiYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYgYTF 
                   
               
               
                 NDISGGYNSSTESFIADPAIiLAHELIHALHGLYGAlGVTYKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNRGQ 
                   
               
               
                 NIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 235 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAELAHELIHALHGLYGAfGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 236 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAfGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYsVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPyKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 237 
               
               
                 NTIGTDPSDFDPPASLENGSfAYYDPNYLTTDAEKDRYLKTTIKLFKRINTS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAfGVTiKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 238 
               
               
                 NTIGTDPSDFDPPdSLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlsVTiKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 239 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAfGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 240 
               
               
                 NTIGTDPSDFDPPASLENGSfAYYDPNYLTTDAEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTiKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNRG 
                   
               
               
                 QNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 241 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENSSYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 242 
               
               
                 NTIGTDPSDFDPPASLdNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMgSGGVYDPSkDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAIiLAHELIHALHGLYGAfGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 243 
               
               
                 NTIGTDPSDFDPPASLdNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMgSGGVYDPSkDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAIiLAHELIHALHGLYGAfGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPdLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 244 
               
               
                 NTIGTDPSDFDPPASLdNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMgSGGVYDPSkDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSgESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIehGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 245 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENyiYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYhSSTESFIADPAIiLAHELIHALHGLYGAlGVTYKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNRGQ 
                   
               
               
                 NIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 246 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENyiYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAIiLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 247 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPdYLTTDAEKDRYLKTTIKLFKRINTS 
                   
               
               
                 NPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIILN 
                   
               
               
                 LLVLGAGPDIFENyiYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYgYTF 
                   
               
               
                 NDISGGYNSSTESFIADPAIiLAHELIHALHGLYGAlGVTYKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNRGQ 
                   
               
               
                 NIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 248 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAELAHELIHALHGLYGAfGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 249 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAfGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYsVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPyKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 250 
               
               
                 NTIGTDPSDFDPPASLENGSfAYYDPNYLTTDAEKDRYLKTTIKLFKRINTS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAfGVTiKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 251 
               
               
                 NTIGTDPSDFDPPdSLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlsVTiKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 252 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAfGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 253 
               
               
                 NTIGTDPSDFDPPASLENGSfAYYDPNYLTTDAEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTiKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNRG 
                   
               
               
                 QNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 254 
               
               
                 NTIGTDPSDFDPPASLdNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMgSGGVYDPSkDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAfGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 255 
               
               
                 NTIGTDPSDFDPPASLdNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMgSGGVYDPSkDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAIiLAHELIHALHGLYGAfGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 256 
               
               
                 NTIGTDPSDFDPPASLdNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMgSGGVYDPSkDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIehGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 257 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENycYPVRKLMDSGGVYDPShDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 258 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENyiYPVRKLMDSGGVYDPShDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGhNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 259 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SiPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENycYPVRKLMDSGGVYDPShDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 260 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSsDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPdLLDDDIYTVSEGFNIGhLAVNNRG 
                   
               
               
                 QNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 261 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENyiYPVRKLMDSGGVYDPShDGFGSINIVTFSPEYaYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 262 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAELAHELIHALHGLYGAfGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYsVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPyKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 263 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDInGGYNSSTESFIADPAELAHELIHALHGLYGAfGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFrKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPyKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 264 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAELAHELIHALHGLYGAfGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYsVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 265 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAELAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYsVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPyKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 266 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTkSFIADPAIiLAHELIHALHGLYGAlGVTcKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIehGFLKVPNLLDDDIYTVSEGFNIGhLAVNNRG 
                   
               
               
                 QNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 267 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSvNIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAELAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYsVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPyKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 268 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAELAHELIHALHGLYGAfGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPgYDINEYKDYFQWKYGLDKNADGSYsVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPyKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 269 
               
               
                 NTIGTDPSDFDPPASLdNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMgSGGVYDPSkDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAfGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 270 
               
               
                 NTIGTDPSDFDPPtSLdNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNvKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMgSGGVYDPSkDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAfGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 271 
               
               
                 NTIGTDPSDFDPPASLdNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMgSGGVYDPSkDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTgSFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIehGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 272 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINT 
                   
               
               
                 SNPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSsDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 273 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINT 
                   
               
               
                 SNPAGEVLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENycYPVRKLMDSGGVYDPShDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 274 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPdYLTTDAEKDRYLKTTIKLFKRINTS 
                   
               
               
                 NPAGEVLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIILN 
                   
               
               
                 LLVLGAGPDIFENycYPVRKLMDSGGVYDPShDGFGSINIVTFSPEYkYTF 
                   
               
               
                 NDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNRG 
                   
               
               
                 QNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 275 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINT 
                   
               
               
                 SNPAGEaLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENycYPVRKLMDSGGVYDPShDGFGSINIVTFSPEYkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 276 
               
               
                 NTIGTDPSDFyPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEaLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIILN 
                   
               
               
                 LLVLGAGPDIFENySYPVRKLMDSGGVYDPSsDGFGSINIVTFSPEYkYTF 
                   
               
               
                 NDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIKhGFLKVPdLLDDDIYTVSEGFNIGhLAVNNRGQ 
                   
               
               
                 NIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 277 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINT 
                   
               
               
                 SNPAGEVLLQEISdAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPShDGFGSINIVTFSPEYkY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYgF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGNLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKawlrKs*KF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 278 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINT 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTcKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFrKVPdLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKcLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEItRNVWIIPERN 
                 279 
               
               
                 TIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGaVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIILN 
                   
               
               
                 LLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkYTF 
                   
               
               
                 NDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTcKETIKVKQAP 
                   
               
               
                 LMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANcEKIATRLSRVN 
                   
               
               
                 SAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEI 
                   
               
               
                 DLANKlKVKCRNTYFIehGFLKVPNLLDDDIYTVSEGFNIGhLAVNNRGQ 
                   
               
               
                 NIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 280 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINT 
                   
               
               
                 SNPAGaVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEhkYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTcKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 281 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgY 
                   
               
               
                 TFNDISGGYNSyTESFIADPAISLAHELIHALHGLYGAlGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDvYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 282 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFyPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAlGVTcKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 283 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYkY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAfGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYsVNENKFNEIYKKLYSF 
                   
               
               
                 TEIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNN 
                   
               
               
                 RGQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 284 
               
               
                 NTIGTDPSDFDPPASLENGSfAYYDPNYLTTDAEKDRYLKTTIKLFKRINS 
                   
               
               
                 NPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFkNySYPVRKLMDSGGVYDPSkDGFGSINIVTFSPEYgYT 
                   
               
               
                 FNDISGGYNSSTESFIADPAISLAHELIHALHGLYGAfGVTYKETIKVKQA 
                   
               
               
                 PLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV 
                   
               
               
                 NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFT 
                   
               
               
                 EIDLANKlKVKCRNTYFIKhGFLKVPNLLDDDIYTVSEGFNIGhLAVNNR 
                   
               
               
                 GQNIKLNlKIIDSIPDKGLVkKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 285 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENSSYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 RVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYS 
                   
               
               
                 FTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVN 
                   
               
               
                 NRGQNIKLNPKIIDSIPDKGLVEKIVKF 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 390 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEALLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENYIYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYGY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAIILAHELIHALHGLYGALGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYGF 
                   
               
               
                 TEIDLANKLKVKCRNTYFIKHGFLKVPNLLDDDIYTVSEGFNIGHLAVN 
                   
               
               
                 NRGQNIKLNLKIIDSIPDKAWLRKS 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 391 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEALLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENYIYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYGY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAIILAHELIHALHGLYGALGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYGF 
                   
               
               
                 TEIDLANKLKVKCRNTYFIKHGFLKVPNLLDDDIYTVSEGFNIGHLAVN 
                   
               
               
                 NRGQNIKLNLKIIDSIPDKAWLRKS 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEESKKYYKAFEIMRNVWIIPER 
                 392 
               
               
                 NTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEALLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIT 
                   
               
               
                 LNLLVLGAGPDIFENYIYPVRKLTDSGGVYDPSNDGFGSINIVTFSPEYG 
                   
               
               
                 YTFNDISGGYNSSTESFIADPAIILAHELIHALHGLYGALGVTYKETIKVK 
                   
               
               
                 QAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLS 
                   
               
               
                 CVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLY 
                   
               
               
                 GFTEIDLANKLKVKCRNTYFIKHGFLKVPNLLDDDIYTVSEGFNIGHLA 
                   
               
               
                 VNNRGQNIKLNLKIIDSIPDKAWLRKS 
                   
               
               
                   
               
               
                 MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPER 
                 393 
               
               
                 NTIGTDPSDFDPPASLENGSSAHYDPNYLTTDAEKDRYLKTTIKLFKRIN 
                   
               
               
                 SNPAGEALLQEISYAKPYLGNEHTPINEFHPGTRTTSVNIKTSTNVKSSIIL 
                   
               
               
                 NLLVLGAGPDIFENYIYPVRKLTDSGGVYDPSNDGFGSINIVTFSPEYGY 
                   
               
               
                 TFNDISGGYNSSTESFIADPAIILAHELIHALHGLYGALGVTYKETIKVKQ 
                   
               
               
                 APLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSR 
                   
               
               
                 VNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYGF 
                   
               
               
                 TEIDLANKLKVKCRNTYFIKHGFLKVPNLLDDDIYTVSEGFNIGHLAVN 
                   
               
               
                 NRGQNIKLNLKIIDSIPDKAWLRKS 
               
               
                   
               
            
           
         
       
     
     This disclosure relates, in part, to the discovery that continuous evolution methods (e.g., PACE) are useful for producing BoNT protease variants that have altered peptide cleaving activities (altered peptide cleaving functions). For example, in some embodiments, a BoNT protease variant as described by the disclosure cleaves a VAMP7 or VAMP8 protein or peptide. In some embodiments, a BoNT protease variant as described by the disclosure cleaves a PTEN protein or peptide. In some embodiments, a BoNT protease variant as described by the disclosure cleaves the target sequence 
     
       
         
           
               
            
               
                 (SEQ ID NO: 314) 
               
               
                 MAILFAVVARGTTILAKHAWCGGNFLEDFERSRAFNFLNEIKKRFQTTYG 
               
               
                   
               
               
                 SRAQTALPYAMNSEFSSVLAAQLKHHSENKGLDKVMETQAQVDELKGIMV 
               
               
                   
               
               
                 RNIDLVAQRGERLELLIDKTENLVDSSVTFKTTSRNLARAMCMKNLKLTI 
               
               
                   
               
               
                 IIIIVSIVFIYIIVSPLCGGFTWPSCVKK 
               
               
                 or 
               
               
                   
               
               
                 (SEQ ID NO: 315) 
               
               
                 GGSGGSGGSKGLDKVMETQAQVDELKGIMVRNIDLVAQRGERLELLIDKT 
               
               
                   
               
               
                 ENLVDSSVTFKTTSRNLARGGSGGSGGS. 
               
            
           
         
       
     
     In some embodiments, a BoNT protease variant as described by the disclosure cleaves the target sequence 
     
       
         
           
               
            
               
                 (SEQ ID NO: 316) 
               
               
                 MTAIIKEIVSRNKRRYQEDGFDLDLTYIYPNIIAMGFPAERLEGVYRNNI 
               
               
                   
               
               
                 DDVVRFLDSKHKNHYKIYNLCAERHYDTAKFNCRVAQYPFEDHNPPQLEL 
               
               
                   
               
               
                 IKPFCEDLDQWLSEDDNHVAAIHCKAGKGRTGVMICAYLLHRGKFLKAQE 
               
               
                   
               
               
                 ALDFYGEVRTRDKKGVTIPSQRRYVYYYSYLLKNHLDYRPVALLFHKMMF 
               
               
                   
               
               
                 ETIPMFSGGTCNPQFVVCQLKVKIYSSNSGPTRREDKFMYFEFPQPLPVC 
               
               
                   
               
               
                 GDIKVEFFHKQNKMLKKDKMFHFWVNTFFIPGPEETSEKVENGSLCDQEI 
               
               
                   
               
               
                 DSICSIERADNDKEYLVLTLTKNDLDKANKDKANRYFSPNFKVKLYFTKT 
               
               
                   
               
               
                 VEEPSNPEASSSTSVTPDVSDNEPDHYRYSDTTDSDPENEPFDEDQHTQI 
               
               
                   
               
               
                 TKV 
               
               
                 or 
               
               
                   
               
               
                 (SEQ ID NO: 317) 
               
               
                 NGSLCDQEIDSICSIERADN. 
               
            
           
         
       
     
     In some embodiments, a BoNT protease variant as described herein cleaves a target sequence selected from: 
                    (SNAP25)       (SEQ ID NO: 318)       MAEDADMRNELEEMQRRADQLADESLESTRRMLQLVEESKDAGIRTLVML               DEQGEQLERIEEGMDQINKDMKEAEKNLTDLGKFCGLCVCPCNKLKSSDA               YKKAWGNNQDGVVASQPARVVDEREQMAISGGFIRRVTNDARENEMDENL               EQVSGIIGNLRHMALDMGNEIDTQNRQIDRIMEKADSNKTRIDEANQRAT               KMLGSG,               (VAMP1)       (SEQ ID NO: 319)       MSAPAQPPAEGTEGTAPGGGPPGPPPNMTSNRRLQQTQAQVEEVVDIIRV               NVDKVLERDQKLSELDDRADALQAGASQFESSAAKLKRKYWWKNCKMMIM               LGAICAIIVVVIVRRG,               (SNAP25; SEQ ID NO: 320)       MGNEIDTQNRQIDRIMEKAD,       and               (VAMP1; SEQ ID NO: 321)       TSNRRLQQTQAQVEEVVDIIRVNVDKVLERDQKLSELDDRADALQAGASQ               FESSAAKLKR,            
and/or cleaves a VAMP7 peptide target sequence, for example,
 
                    (SEQ ID NO: 322)       MAILFAVVARGTTILAKHAWCGGNFLEDFERSRAFNFLNEIKKRFQTTYG               SRAQTALPYAMNSEFSSVLAAQLKHHSENKGLDKVMETQAQVDELKGIMV               RNIDLVAQRGERLELLIDKTENLVDSSVTFKTTSRNLARAMCMKNLKLTI               IIIIVSIVFIYIIVSPLCGGFTWPSCVKK,       or               (SEQ ID NO: 323)       GGSGGSGGSKGLDKVMETQAQVDELKGIMVRNIDLVAQRGERLELLIDKT               ENLVDSSVTFKTTSRNLARGGSGGSGGS.            
In some embodiments, a BoNT protease variant cleaves a target peptide (e.g., VAMP7, VAMP8, PTEN, etc.) with higher activity than a wild-type BoNT protease. A BoNT protease variant that cleaves a target peptide (e.g., VAMP7, VAMP8, PTEN, etc.) with higher activity can have an increase in catalytic efficiency ranging from about 1.1-fold, about 1.5-fold, 2-fold to about 100-fold, about 5-fold to about 50-fold, or about 10-fold to about 40-fold, relative to the catalytic efficiency of the wild-type BoNT protease from which the BoNT protease variant was derived. In some embodiments, a BoNT protease variant described herein cleaves a target peptide (e.g., VAMP7, VAMP8, PTEN, etc.) with about 1% to about 100% (e.g., about 1%, 2%, 5%, 10%, 20%, 50%, 80%, 90%, 100%) of the catalytic efficiency with which wild-type BoNT cleaves its native substrate (e.g., SNAP25, VAMP1, etc.). Catalytic efficiency can be measured or determined using any suitable method known in the art, for example using the methods described in Harris et al. (2009)  Methods Enzymol.  463; 57-71.
 
     Generally, the evolution of proteases with altered specificity has focused exclusively on the destruction of therapeutically relevant extracellular proteins. However BoNTs provide a built-in cytosolic delivery mechanism, and thus are able, in some embodiments, to degrade intracellular targets. For example, in some embodiments, a BoNT protease variant as described herein comprises one or more protein domains that facilitate transport of the protease across a cellular membrane. In some embodiments, the one or more protein domains that facilitate transport across the membrane are selected from a BoNT HC, a BoNT HCC domain, and a BoNT HCN domain. In some embodiments, BoNT protease variants described by the disclosure are capable of crossing the cellular membrane and entering the intracellular environment of neuronal cell types. 
     Some aspects of this disclosure provide methods for using a protease provided herein. In some embodiments, such methods include contacting a protein comprising a protease target cleavage sequence, for example ex vivo, in vitro, or in vivo (e.g., in a subject), with the protease. In some embodiments, the protein contacted with the protease is a therapeutic target. In some embodiments, the therapeutic target is VAMP7. Generally, VAMP7 is an intracellular, soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) family protein that mediates the fusion of transport vesicles to their target membrane. Without wishing to be bound by any particular theory, VAMP7 functions as a mediator of MT1-MMP secretion during tumor invasion and granzyme B and perform secretion, for example, during organ transplantation. Accordingly, in some aspects, the disclosure provides methods of decreasing VAMP7 activity in a cell, the method comprising contacting the cell with, or introducing into the intracellular environment of the cell, a variant BoNT protease as described herein. 
     In some embodiments, the therapeutic target is PTEN. Generally, PTEN is an intracellular protein comprising a tensin domain and a phosphatase domain that functions as a tumor suppressor. PTEN has also been observed to mediate ischemic neuronal damage after a stroke. Accordingly, in some aspects, the disclosure provides methods of decreasing PTEN activity in a cell, the method comprising contacting the cell with, or introducing into the intracellular environment, a BoNT protease variant as described herein (e.g., a BoNT E variant). 
     In some embodiments, the cell (or intracellular environment) is characterized by increased or undesired activity of a target protein (e.g., VAMP7, VAMP8, PTEN, etc.) relative to a normal cell or extracellular environment (e.g., a healthy cell, or extracellular environment, not characterized by increased activity of the target protein). In some embodiments, increased activity of a target protein (e.g., VAMP7, VAMP8, PTEN, etc.) occurs when, in a cell, the activity of the target protein is about 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 25-fold, 50-fold, 100-fold, 500-fold, or 1000-fold over activity of the target protein in a normal healthy cell, or extracellular environment. In some embodiments, a cell characterized by increased expression of a target protein (e.g., VAMP7, VAMP8, PTEN, etc.) is derived from a subject (e.g., a mammalian subject, such as a human or mouse) that has or is suspected of having a disease associated with increased activity of the target gene, for example, cancer in the context of VAMP7 overexpression or increased activity. 
     In some embodiments, the methods provided herein comprise contacting (e.g., cleaving) the target protein (e.g., VAMP7, PTEN, etc., or a protein comprising a peptide comprising an amino acid sequence set forth in SEQ ID NOs.: 314-323 (e.g., VAMP7, PTEN, etc.)) with a BoNT protease variant described herein in vitro. In some embodiments, the methods provided herein comprise contacting the target protein with the protease variant described herein in vivo. In some embodiments, the methods provided herein comprise contacting the target protein (e.g., VAMP7, PTEN, etc., or a protein comprising a peptide comprising an amino acid sequence set forth in SEQ ID NOs.: 314-323 (e.g., VAMP7, PTEN)) with a BoNT protease variant described herein in an intracellular environment. In some embodiments, the methods provided herein comprise contacting the target protein (e.g., VAMP7, PTEN, etc., or a protein comprising a peptide comprising an amino acid sequence set forth in SEQ ID NOs.: 314-323 (e.g. VAMP7, PTEN)) with a BoNT protease variant in a subject, e.g., by administering the protease to the subject, either locally or systemically. In some such embodiments, the protease variant is administered to the subject in an amount effective to result in a measurable decrease in the level of full-length (or functional) target protein (e.g., VAMP7, PTEN, etc.) in the subject, or in a measurable increase in the level of a cleavage product generated by the protease variant upon cleavage of the target protein. 
     Engineering of BoNT Protease Variants Using PACE 
     Some aspects of this disclosure provide methods for evolving a BoNT protease. In some embodiments, a method of evolving a protease is provided that comprises (a) contacting a population of host cells with a population of vectors comprising a gene encoding a protease to be evolved. The vectors are typically deficient in at least one gene required for the transfer of the phage vector from one cell to another, e.g., a gene required for the generation of infectious phage particles. In some embodiments of the provided methods, (1) the host cells are amenable to transfer of the vector; (2) the vector allows for expression of the protease in the host cell, can be replicated by the host cell, and the replicated vector can transfer into a second host cell; and (3) the host cell expresses a gene product encoded by the at least one gene for the generation of infectious phage particles (a) in response to the activity of the protease, and the level of gene product expression depends on the activity of the protease. The methods of protease evolution provided herein typically comprise (b) incubating the population of host cells under conditions allowing for mutation of the gene encoding the protease, and the transfer of the vector comprising the gene encoding the protease of interest from host cell to host cell. The host cells are removed from the host cell population at a certain rate, e.g., at a rate that results in an average time a host cell remains in the cell population that is shorter than the average time a host cell requires to divide, but long enough for the completion of a life cycle (uptake, replication, and transfer to another host cell) of the vector. The population of host cells is replenished with fresh host cells that do not harbor the vector. In some embodiments, the rate of replenishment with fresh cells substantially matches the rate of removal of cells from the cell population, resulting in a substantially constant cell number or cell density within the cell population. The methods of protease evolution provided herein typically also comprise (c) isolating a replicated vector from the host cell population of step (b), wherein the replicated vector comprises a mutated version of the gene encoding the protease. 
     Some embodiments provide a continuous evolution system, in which a population of viral vectors, e.g., M13 phage vectors, comprising a gene encoding a protease of interest to be evolved replicates in a flow of host cells, e.g., a flow through a lagoon, wherein the viral vectors are deficient in a gene encoding a protein that is essential for the generation of infectious viral particles, and wherein that gene is in the host cell under the control of a conditional promoter, the activity of which depends on the activity of the protease of interest. In some embodiments, transcription from the conditional promoter may be activated by cleavage of a fusion protein comprising a transcription factor and an inhibitory protein fused to the transcriptional activator via a linker comprising a target site of the protease. 
     Some embodiments of the protease PACE technology described herein utilize a “selection phage,” a modified phage that comprises a gene of interest to be evolved and lacks a full-length gene encoding a protein required for the generation of infectious phage particles. In some such embodiments, the selection phage serves as the vector that replicates and evolves in the flow of host cells. For example, some M13 selection phages provided herein comprise a nucleic acid sequence encoding a protease to be evolved, e.g., under the control of an M13 promoter, and lack all or part of a phage gene encoding a protein required for the generation of infectious phage particles, e.g., gI, gII, gIII, gIV, gV, gVI, gVII, gVIII, gIX, or gX, or any combination thereof. For example, some M13 selection phages provided herein comprise a nucleic acid sequence encoding a protease to be evolved, e.g., under the control of an M13 promoter, and lack all or part of a gene encoding a protein required for the generation of infectious phage particles, e.g., the gIII gene encoding the pIII protein. 
     One prerequisite for evolving proteases with a desired activity is to provide a selection system that confers a selective advantage to mutated protease variants exhibiting such an activity. The expression systems and fusion proteins comprising transcriptional activators in an inactive form that are activated by protease activity thus constitute an important feature of some embodiments of the protease PACE technology provided herein. 
     In some embodiments, the transcriptional activator directly drives transcription from a target promoter. For example, in some such embodiments, the transcriptional activator may be an RNA polymerase. Suitable RNA polymerases and promoter sequences targeted by such RNA polymerases are well known to those of skill in the art. Exemplary suitable RNA polymerases include, but are not limited to, T7 polymerases (targeting T7 promoter sequences) and T3 RNA polymerases (targeting T3 promoter sequences). Additional suitable RNA polymerases will be apparent to those of skill in the art based on the instant disclosure, which is not limited in this respect. 
     In some embodiments, the transcriptional activator does not directly drive transcription, but recruits the transcription machinery of the host cell to a specific target promoter. Suitable transcriptional activators, such as, for example, Gal4 or fusions of the transactivation domain of the VP16 transactivator with DNA-binding domains, will be apparent to those of skill in the art based on the instant disclosure, and the disclosure is not limited in this respect. 
     In some embodiments, it is advantageous to link protease activity to enhanced phage packaging via a transcriptional activator that is not endogenously expressed in the host cells in order to minimize leakiness of the expression of the gene required for the generation of infectious phage particles through the host cell basal transcription machinery. For example, in some embodiments, it is desirable to drive expression of the gene required for the generation of infectious phage particles from a promoter that is not or is only minimally active in host cells in the absence of an exogenous transcriptional activator, and to provide the exogenous transcriptional activator, such as, for example, T7 RNA polymerase, as part of the expression system linking protease (e.g., BoNT protease variant) activity to phage packaging efficiency. In some embodiments, the at least one gene for the generation of infectious phage particles is expressed in the host cells under the control of a promoter activated by the transcriptional activator, for example, under the control of a T7 promoter if the transcriptional activator is T7 RNA polymerase, and under the control of a T3 promoter if the transcriptional activator is T3 polymerase, and so on. 
     In some embodiments, the transcriptional activator is fused to an inhibitor that either directly inhibits or otherwise hinders the transcriptional activity of the transcriptional activator, for example, by directly interfering with DNA binding or transcription, by targeting the transcriptional activator for degradation through the host cells protein degradation machinery, or by directing export from the host cell or localization of the transcriptional activator into a compartment of the host cell in which it cannot activate transcription from its target promoter. In some embodiments, the inhibitor is fused to the transcriptional activator&#39;s N-terminus. In other embodiments, it is fused to the activator&#39;s C-terminus. 
     In some embodiments, the protease evolution methods provided herein comprise an initial or intermittent phase of diversifying the population of vectors by mutagenesis, in which the cells are incubated under conditions suitable for mutagenesis of the gene encoding the protease in the absence of stringent selection or in the absence of any selection for evolved protease variants that have acquired a desired activity. Such low-stringency selection or no selection periods may be achieved by supporting expression of the gene for the generation of infectious phage particles in the absence of desired protease activity, for example, by providing an inducible expression construct comprising a gene encoding the respective packaging protein under the control of an inducible promoter and incubating under conditions that induce expression of the promoter, e.g., in the presence of the inducing agent. Suitable inducible promoters and inducible expression systems are described herein and in International PCT Application, PCT/US2011/066747, filed Dec. 22, 2011, published as WO 2012/088381 on Jun. 28, 2012; and U.S. Application, U.S. Ser. No. 13/922,812, filed Jun. 20, 2013; International PCT Application, PCT/US2015/057012, filed on Oct. 22, 2015, published as WO 2016/077052; and, PCT/US2016/027795, filed on Apr. 15, 2016, published as WO 2016/168631, the entire contents of each of which are incorporated herein by reference. Additional suitable promoters and inducible gene expression systems will be apparent to those of skill in the art based on the instant disclosure. In some embodiments, the method comprises a phase of stringent selection for a mutated protease version. If an inducible expression system is used to relieve selective pressure, the stringency of selection can be increased by removing the inducing agent from the population of cells in the lagoon, thus turning expression from the inducible promoter off, so that any expression of the gene required for the generation of infectious phage particles must come from the protease activity-dependent expression system. 
     One aspect of the PACE protease evolution methods provided herein is the mutation of the initially provided vectors encoding a protease of interest. In some embodiments, the host cells within the flow of cells in which the vector replicates are incubated under conditions that increase the natural mutation rate. This may be achieved by contacting the host cells with a mutagen, such as certain types of radiation or to a mutagenic compound, or by expressing genes known to increase the cellular mutation rate in the cells. Additional suitable mutagens will be known to those of skill in the art, and include, without limitation, those described in International PCT Application, PCT/US2011/066747, filed Dec. 22, 2011, published as WO 2012/088381 on Jun. 28, 2012; and U.S. Application, U.S. Ser. No. 13/922,812, filed Jun. 20, 2013; International PCT Application, PCT/US2015/057012, filed on Oct. 22, 2015, published as WO 2016/077052; and, PCT/US2016/027795, filed on Apr. 15, 2016, published as WO 2016/168631, the entire contents of each of which are incorporated herein by reference and the disclosure is not limited in this respect. 
     In some embodiments, the host cells comprise the accessory plasmid encoding the at least one gene for the generation of infectious phage particles, e.g., of the M13 phage, encoding the protease to be evolved and a helper phage, and together, the helper phage and the accessory plasmid comprise all genes required for the generation of infectious phage particles. Accordingly, in some such embodiments, variants of the vector that do not encode a protease variant that can untether the inhibitor from the transcriptional activator will not efficiently be packaged, since they cannot effect an increase in expression of the gene required for the generation of infectious phage particles from the accessory plasmid. On the other hand, variants of the vector that encode a protease variant that can efficiently cleave the inhibitor from the transcriptional activator will effect increased transcription of the at least one gene required for the generation of infectious phage particles from the accessory plasmid and thus be efficiently packaged into infectious phage particles. 
     In some embodiments, the protease PACE methods provided herein further comprises a negative selection for undesired protease activity in addition to the positive selection for a desired protease activity. Such negative selection methods are useful, for example, in order to maintain protease specificity when increasing the cleavage efficiency of a protease directed towards a specific target site. This can avoid, for example, the evolution of proteases that show a generally increased protease activity, including an increased protease activity towards off-target sites, which is generally undesired in the context of therapeutic proteases. 
     In some embodiments, negative selection is applied during a continuous evolution process as described herein, by penalizing the undesired activities of evolved proteases. This is useful, for example, if the desired evolved protease is an enzyme with high specificity for a target site, for example, a protease with altered, but not broadened, specificity. In some embodiments, negative selection of an undesired activity, e.g., off-target protease activity, is achieved by causing the undesired activity to interfere with pIII production, thus inhibiting the propagation of phage genomes encoding gene products with an undesired activity. In some embodiments, expression of a dominant-negative version of pIII or expression of an antisense RNA complementary to the gIII RBS and/or gIII start codon is linked to the presence of an undesired protease activity. Suitable negative selection strategies and reagents useful for negative selection, such as dominant-negative versions of M13 pIII, are described herein and in International PCT Application, PCT/US2011/066747, filed Dec. 22, 2011, published as WO 2012/088381 on Jun. 28, 2012; and U.S. Application, U.S. Ser. No. 13/922,812, filed Jun. 20, 2013; International PCT Application, PCT/US2015/057012, filed on Oct. 22, 2015, published as WO 2016/077052; and, PCT/US2016/027795, filed on Apr. 15, 2016, published as WO 2016/168631, the entire contents of each of which are incorporated herein by reference. 
     In some embodiments, counter-selection against activity on non-target substrates is achieved by linking undesired evolved protease activities to the inhibition of phage propagation. In some embodiments, a dual selection strategy is applied during a continuous evolution experiment, in which both positive selection and negative selection constructs are present in the host cells. In some such embodiments, the positive and negative selection constructs are situated on the same plasmid, also referred to as a dual selection accessory plasmid. 
     One advantage of using a simultaneous dual selection strategy is that the selection stringency can be fine-tuned based on the activity or expression level of the negative selection construct as compared to the positive selection construct. Another advantage of a dual selection strategy is that the selection is not dependent on the presence or the absence of a desired or an undesired activity, but on the ratio of desired and undesired activities, and, thus, the resulting ratio of pIII and pIII-neg that is incorporated into the respective phage particle. 
     For example, in some embodiments, the host cells comprise an expression construct encoding a dominant-negative form of the at least one gene for the generation of infectious phage particles, e.g., a dominant-negative form of the pIII protein (pIII-neg), under the control of an inducible promoter that is activated by a transcriptional activator other than the transcriptional activator driving the positive selection system. Expression of the dominant-negative form of the gene diminishes or completely negates any selective advantage an evolved phage may exhibit and thus dilutes or eradicates any variants exhibiting undesired activity from the lagoon. 
     For example, if the positive selection system comprises a T7 promoter driving the expression of the at least one gene for the generation of infectious phage particles, and a T7 RNA polymerase fused to a T7-RNA polymerase inhibitor via a linker comprising a protease target site that is cleaved by a desired protease activity, the negative selection system should be a non-T7 based system. For example, in some such embodiments, the negative selection system could be based on T3 polymerase activity, e.g., in that it comprises a T3 promoter driving the expression of a dominant-negative form of the at least one gene for the generation of infectious phage particles, and a T3 RNA polymerase fused to a T3-RNA polymerase inhibitor via a linker comprising a protease target site that is cleaved by an undesired protease activity. In some embodiments, the negative selection polymerase is a T7 RNA polymerase gene comprising one or more mutations that render the T7 polymerase able to transcribe from the T3 promoter but not the T7 promoter, for example: N67S, R96L, K98R, H176P, E207K, E222K, T375A, M4011, G675R, N748D, P759L, A798S, A819T, etc. In some embodiments the negative selection polymerase may be fused to a T7-RNA polymerase inhibitor via a linker comprising a protease target site that is cleaved by an undesired protease activity. When used together, such positive-negative PACE selection results in the evolution of proteases that exhibit the desired activity but not the undesired activity. In some embodiments, the undesired function is cleavage of an off-target protease cleavage site. In some embodiments, the undesired function is cleavage of the linker sequence of the fusion protein outside of the protease cleavage site. 
     Some aspects of this invention provide or utilize a dominant negative variant of pIII (pIII-neg). These aspects are based on the recognition that a pIII variant that comprises the two N-terminal domains of pIII and a truncated, termination-incompetent C-terminal domain is not only inactive but is a dominant-negative variant of pIII. A pIII variant comprising the two N-terminal domains of pIII and a truncated, termination-incompetent C-terminal domain was described in Bennett, N. J.; Rakonjac, J., Unlocking of the filamentous bacteriophage virion during infection is mediated by the C domain of pIII.  Journal of Molecular Biology  2006, 356 (2), 266-73; the entire contents of which are incorporated herein by reference. The dominant negative property of such pIII variants has been described in more detail in PCT Application PCT/US2011/066747, published as WO 2012/088381 on Jun. 28, 2012, the entire contents of which are incorporated herein by reference. 
     The pIII-neg variant as provided in some embodiments herein is efficiently incorporated into phage particles, but it does not catalyze the unlocking of the particle for entry during infection, rendering the respective phage noninfectious even if wild type pIII is present in the same phage particle. Accordingly, such pIII-neg variants are useful for devising a negative selection strategy in the context of PACE, for example, by providing an expression construct comprising a nucleic acid sequence encoding a pIII-neg variant under the control of a promoter comprising a recognition motif, the recognition of which is undesired. In other embodiments, pIII-neg is used in a positive selection strategy, for example, by providing an expression construct in which a pIII-neg encoding sequence is controlled by a promoter comprising a nuclease target site or a repressor recognition site, the recognition of either one is desired. 
     In some embodiments, a protease PACE experiment according to methods provided herein is run for a time sufficient for at least 10, at least 20, at least 30, at least 40, at least 50, at least 100, at least 200, at least 300, at least 400, at least, 500, at least 600, at least 700, at least 800, at least 900, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2500, at least 3000, at least 4000, at least 5000, at least 7500, at least 10000, or more consecutive viral life cycles. In certain embodiments, the viral vector is an M13 phage, and the length of a single viral life cycle is about 10-20 minutes. 
     In some embodiments, the host cells are contacted with the vector and/or incubated in suspension culture. For example, in some embodiments, bacterial cells are incubated in suspension culture in liquid culture media. Suitable culture media for bacterial suspension culture will be apparent to those of skill in the art, and the invention is not limited in this regard. See, for example,  Molecular Cloning: A Laboratory Manual,  2nd Ed., ed. by Sambrook, Fritsch, and Maniatis (Cold Spring Harbor Laboratory Press: 1989); Elizabeth Kutter and Alexander Sulakvelidze:  Bacteriophages: Biology and Applications . CRC Press; 1 st  edition (December 2004), ISBN: 0849313368; Martha R. J. Clokie and Andrew M. Kropinski:  Bacteriophages: Methods and Protocols, Volume  1 : Isolation, Characterization, and Interactions  ( Methods in Molecular Biology ) Humana Press; 1 st  edition (December, 2008), ISBN: 1588296822; Martha R. J. Clokie and Andrew M. Kropinski:  Bacteriophages: Methods and Protocols, Volume  2 : Molecular and Applied Aspects  ( Methods in Molecular Biology ) Humana Press; 1 st  edition (December 2008), ISBN: 1603275649; all of which are incorporated herein in their entirety by reference for disclosure of suitable culture media for bacterial host cell culture). 
     The protease PACE methods provided herein are typically carried out in a lagoon. Suitable lagoons and other laboratory equipment for carrying out protease PACE methods as provided herein have been described in detail elsewhere. See, for example, International PCT Application, PCT/US2011/066747, published as WO2012/088381 on Jun. 28, 2012, the entire contents of which are incorporated herein by reference. In some embodiments, the lagoon comprises a cell culture vessel comprising an actively replicating population of vectors, for example, phage vectors comprising a gene encoding the protease of interest, and a population of host cells, for example, bacterial host cells. In some embodiments, the lagoon comprises an inflow for the introduction of fresh host cells into the lagoon and an outflow for the removal of host cells from the lagoon. In some embodiments, the inflow is connected to a turbidostat comprising a culture of fresh host cells. In some embodiments, the outflow is connected to a waste vessel, or a sink. In some embodiments, the lagoon further comprises an inflow for the introduction of a mutagen into the lagoon. In some embodiments that inflow is connected to a vessel holding a solution of the mutagen. In some embodiments, the lagoon comprises an inflow for the introduction of an inducer of gene expression into the lagoon, for example, of an inducer activating an inducible promoter within the host cells that drives expression of a gene promoting mutagenesis (e.g., as part of a mutagenesis plasmid), as described in more detail elsewhere herein. In some embodiments, that inflow is connected to a vessel comprising a solution of the inducer, for example, a solution of arabinose. 
     In some embodiments, a PACE method as provided herein is performed in a suitable apparatus as described herein. For example, in some embodiments, the apparatus comprises a lagoon that is connected to a turbidostat comprising a host cell as described herein. In some embodiments, the host cell is an  E. coli  host cell. In some embodiments, the host cell comprises an accessory plasmid as described herein, a helper plasmid as described herein, a mutagenesis plasmid as described herein, and/or an expression construct encoding a fusion protein as described herein, or any combination thereof. In some embodiments, the lagoon further comprises a selection phage as described herein, for example, a selection phage encoding a protease of interest. In some embodiments, the lagoon is connected to a vessel comprising an inducer for a mutagenesis plasmid, for example, arabinose. In some embodiments, the host cells are  E. coli  cells comprising the F′ plasmid, for example, cells of the genotype F′proA + B + Δ(lacIZY) zzf::Tn10(TetR)/endA1 recA1 galE15 galK16 nupG rpsL ΔlacIZYA araD139 Δ(ara,leu)7697 mcrA Δ(mrr-hsdRMS-mcrBC) proBA::pir116 λ − . 
     Some aspects of this invention relate to host cells for continuous evolution processes as described herein. In some embodiments, a host cell is provided that comprises at least one viral gene encoding a protein required for the generation of infectious viral particles under the control of a conditional promoter, and a fusion protein comprising a transcriptional activator targeting the conditional promoter and fused to an inhibitor via a linker comprising a protease cleavage site. For example, some embodiments provide host cells for phage-assisted continuous evolution processes, wherein the host cell comprises an accessory plasmid comprising a gene required for the generation of infectious phage particles, for example, M13 gIII, under the control of a conditional promoter, as described herein. In some embodiments, the host cells comprises an expression construct encoding a fusion protein as described herein, e.g., on the same accessory plasmid or on a separate vector. In some embodiments, the host cell further provides any phage functions that are not contained in the selection phage, e.g., in the form of a helper phage. In some embodiments, the host cell provided further comprises an expression construct comprising a gene encoding a mutagenesis-inducing protein, for example, a mutagenesis plasmid as provided herein. 
     In some embodiments, modified viral vectors are used in continuous evolution processes as provided herein. In some embodiments, such modified viral vectors lack a gene required for the generation of infectious viral particles. In some such embodiments, a suitable host cell is a cell comprising the gene required for the generation of infectious viral particles, for example, under the control of a constitutive or a conditional promoter (e.g., in the form of an accessory plasmid, as described herein). In some embodiments, the viral vector used lacks a plurality of viral genes. In some such embodiments, a suitable host cell is a cell that comprises a helper construct providing the viral genes required for the generation of infectious viral particles. A cell is not required to actually support the life cycle of a viral vector used in the methods provided herein. For example, a cell comprising a gene required for the generation of infectious viral particles under the control of a conditional promoter may not support the life cycle of a viral vector that does not comprise a gene of interest able to activate the promoter, but it is still a suitable host cell for such a viral vector. 
     In some embodiments, the host cell is a prokaryotic cell, for example, a bacterial cell. In some embodiments, the host cell is an  E. coli  cell. In some embodiments, the host cell is a eukaryotic cell, for example, a yeast cell, an insect cell, or a mammalian cell. The type of host cell, will, of course, depend on the viral vector employed, and suitable host cell/viral vector combinations will be readily apparent to those of skill in the art. 
     In some embodiments, the viral vector is a phage and the host cell is a bacterial cell. In some embodiments, the host cell is an  E. coli  cell. Suitable  E. coli  host strains will be apparent to those of skill in the art, and include, but are not limited to, New England Biolabs (NEB) Turbo, Top10F′, DH12S, ER2738, ER2267, and XL1-Blue MRF′. These strain names are art recognized and the genotype of these strains has been well characterized. It should be understood that the above strains are exemplary only and that the invention is not limited in this respect. 
     In some PACE embodiments, for example, in embodiments employing an M13 selection phage, the host cells are  E. coli  cells expressing the Fertility factor, also commonly referred to as the F factor, sex factor, or F-plasmid. The F-factor is a bacterial DNA sequence that allows a bacterium to produce a sex pilus necessary for conjugation and is essential for the infection of  E. coli  cells with certain phage, for example, with M13 phage. For example, in some embodiments, the host cells for M13-PACE are of the genotype F′proA + B + Δ(lacIZY) zzf::Tn10(TetR)/endA1 recA1 galE15 galK16 nupG rpsL ΔlacIZYA araD139 Δ(ara,leu)7697 mcrA Δ(mrr-hsdRMS-mcrBC) proBA::pir116 λ. 
     Some of the embodiments, advantages, features, and uses of the technology disclosed herein will be more fully understood from the Examples below. The Examples are intended to illustrate some of the benefits of the present disclosure and to describe particular embodiments, but are not intended to exemplify the full scope of the disclosure and, accordingly, do not limit the scope of the disclosure. 
     EXAMPLES 
     Example 1 
     Among the most notable protein systems capable of cytoplasmic delivery are the Clostridial neurotoxins, which include seven serologically distinct Botulinum neurotoxins (BoNT A-G) and a single Tetanus Syntaxin neurotoxin (TeNT). These modular proteins are expressed as a single 150 kDa single protein, which is proteolyzed into two components, a 100 kDa “heavy chain” (HC) and 50 kDa “light chain” (LC), which remain associated through a single disulfide linkage. The BoNT HC is further subdivided into a HCC binding domain, responsible for binding to cholinergic motor nerve terminals, and HCN translocation domain. Upon binding and endocytosis by neurons, endosomal acidification drives a structural reorganization of the translocation domain (HCN) which transports the tethered LC zinc metalloprotease (BoNT LC) to the neurotoxins cytoplasm ( FIG. 1 ). The protease is then released after reduction of the disulfide linkage, and proceeds to block neurotransmitter release by proteolytic cleavage of vesicular membrane fusion (SNARE) proteins, thereby causing paralysis ( FIG. 1 ). The catalytic nature of this event, and the long intracellular lifetime of these proteases results in BoNT neurotoxins being among the most potent neurotoxic agents known. 
     Two BoNT serotypes (BoNT A and BoNT B) are approved for several therapeutic applications, including cervical dystonia, blepharospasm, and spasticity. Therapies based on wild-type BoNTs are notable for their exquisite selectivity for both neuronal cells and for specific SNARE proteins, but these characteristics also limit the broader application of these proteins for modulating cellular chemistry. While notable advances have been made in retargeting BoNT toxins to novel neuronal subtypes or to non-neuronal cells, the preferential SNARE substrates for BoNT LC proteases function primarily in neuronal signaling. Efforts to modify the activity of the BoNT LC protease to overcome this restriction have met with much more limited success, and the inaccessibility of new intracellular targets for BoNT LC proteases remains the primary barrier in extending BoNT-type therapeutic strategies. 
     Proteolysis represents one of the most common post-translational modifications, and the ability to redirect selective BoNT LC proteases to novel substrates within the cell would constitute a powerful therapeutic tool. Reprogramming proteases to permanently modify disease-associated proteins has been a longstanding goal within the protein engineering community, but the challenges of altering protease activity and selectivity have limited the broad utilization of proteases in medicine. Substantial efforts to reprogram proteases have demonstrated the feasibility of tuning protease activity and selectivity by directed evolution. However, because most reported successes to date have been limited to single-residue specificity changes, commonly with greatly impaired levels of activity, therapeutic applications of proteases have thus far been limited to proteases that perform native functions in vivo. BoNT proteases are not exempt from to this trend. Only limited success in protease engineering successes have been reported, and have thus far been unable to substantially broaden the applications of BoNT therapies. 
     Phage assisted continuous evolution (PACE) has recently emerged as a powerful strategy for the development of new activity in proteins, including new promoter selectivity in RNA polymerases, new binding selectivity among protein-DNA interactions, new binding activity in protein-protein interactions, and drug resistance among viral proteases. The PACE selection is built around a modified M13 filamentous bacteriophage, which is unable to infect new hosts due to a lack of the essential gene III and its product, the coat protein pIII ( FIG. 2 ). This essential gene has been moved to  E. coli  host cells, and its transcription is placed under the control of a specified activity of interest. In order to select for improved activity, the protein to be evolved is carried by the phage (selection phage) and is expressed upon bacterial host infection. Because pIII is essential for this infection process, only phage carrying active library members will be able to produce infectious progeny, and their infectivity increases in proportion to the activity of the encoded protein. The selection itself takes place in a dynamic lagoon environment undergoing constant dilution with new  E. coli  cells and with constant outflow to removing inactive phage library members, thereby allowing only phage containing the most active protein variants to persist. The ability to perform and control in situ mutagenesis, selection, and replication in PACE offers many advantages over traditional directed evolution approaches. Generally, PACE enables much faster selections with large gene libraries, resulting in a ˜100-fold increase in the rate of protein evolution efforts, as hundreds of rounds of evolution can be performed in the course of a one-week PACE experiment with minimal researcher intervention. Improvements in controlling selection stringency and mutagenesis rate allow PACE to cover a vast region of a given protein&#39;s adaptive landscape, making it a powerful approach that can surpass rational engineering efforts in accessing novel protein activities. While highly efficient, PACE selection requires linkage of protein activity to transcription of gene-III to drive the selection, for example, a protease-activated T7 RNA polymerase (referred to hereafter as T7-PAP) that is capable of driving the evolution of proteases including BoNT protease and the Hepatitis C viral (HCV) NS3/4Aprotease. In this example platform, gene-III is placed under the control of the T7 promoter, and the cognate T7 polymerase is expressed as a fusion construct with T7 lysozyme with a protease-sensitive linker. Because T7 lysozyme is a natural inhibitor of T7 RNAP, T7pos is unable to transcribe pIII until the linking domain is proteolytically cleaved. ( FIG. 3 ). Upon hydrolysis, the T7 polymerase becomes activated, and is able to transcribe gene-III from the T7 promoter in the accessory plasmid. Importantly, the linker region in T7pos can be varied to accommodate a variety of cleavage sequence, and is sensitive to the intrinsic selectivity of a theco-expressed protease. 
     Eukaryotic organisms, including humans, rely heavily on lipid membranes to compartmentalize and control biochemical processes. Complementary sets of SNARE proteins control trafficking of vesicular organelles, and are the primary mechanism by which cells catalyze membrane fusion events that lead to secretion, autophagy, and membrane remodeling. Despite the obvious potential for controlling signal networks through control over these processes, this therapeutic strategy remains underexplored, and the only small-molecule secretion inhibitors known have limited clinical viability due to low specificity and high toxicity. BoNT neurotoxins, offer a solution to this limitation, but require expansion of their activity to extend the scope of this therapeutic strategy. BoNT B, BoNT D, and BoNT F LC proteases selectively hydrolyze vesicle associated membrane protein-1 (VAMP1) and VAMP2, thereby blocking neurotransmitter secretion. However, several other VAMP family members such as VAMP7 (TI-VAMP), mediate important cellular events including autophagy, plasma membrane remodeling, and secretion, but are not cleaved by BoNT proteases. VAMP7 is the primary v-SNARE responsible for MT1-MMP secretion during tumor cell invasion, and also mediates granzyme B and perforin secretion during natural killer mediated cell death, a major hurdle in transplantation efforts. Given its close relationship to natural BoNT substrates, VAMP7 represents an ideal first target for simultaneously expanding BoNT LC protease activity for biomacromolecular modulation of intracellular chemistry, and broadening the applications for targeted inhibitors of trafficking and secretion. 
     In addition to providing an engineered BoNT protease with potential therapeutic relevance, this example establishes a foundation for extending intracellular biological treatments using the BoNT platform. 
     Evolution of BoNTs Using PACE 
     The first-generation T7-PAP construct possessed only a short (seven amino acid) substrate sequence, and was flanked by flexible linkers to allow T7 lysozyme binding in the unhydrolyzed state ( FIG. 4B ). However, this condensed cleavage sequence differs dramatically from the extended substrate recognition site in BoNT proteases. VAMP-cleaving BoNT proteases require a sequence of approximately 30 amino acids to perform efficient hydrolysis in isolated peptides, and thus it becomes essential to establish whether an elongated substrate sequence can be incorporated into the T7-PAP with retention of protease-dependent transcriptional activity. In order to validate expression and proteolytic activity of recombinantly expressed BoNT LC proteases (hereafter annotated to as BoNT N, where N denotes the neurotoxin serotype), selection phagemids containing each of the three VAMP1-cleaving BoNT serotypes (BoNT LC B, D, and F) were obtained. These proteases were assayed against VAMP1 and VAMP2-linked T7-PAP constructs (T7-PAP(VAMP1)/T7-PAP(VAMP2)) in a coupled luciferase assay ( FIG. 4C ). The resulting data demonstrate that BoNT F is expressed upon M13 phage infection of  E. coli , and can drive protease-dependent transcription from the SNARE-derived T7-PAPs. Selection phage were then submitted to a PACE on the T7-PAPVAMP1 substrate, yielding a mutation (S166Y) that dramatically improves cleavage activity on the natural substrate ( FIG. 4D , third column). These results demonstrate that PACE selection can be applied to BoNT protease evolution using a SNARE-protein adapted T7-PAP. 
     Previous characterization of BoNT F protease promiscuity has revealed a collection of residues in VAMP1 that are important for efficient cleavage activity, and a number of these sites coincide with non-conserved amino acids in VAMP7 ( FIG. 4A ). These residues (as shows in  FIG. 4B ) were selected as primary targets for assessing initial PACE trajectories for BoNT protease reprogramming. A panel of AP&#39;s encoding T7-PAPs containing VAMP1 single-mutant substrates for VAMP7-targeted evolution was produced, and the activity of BoNT F(wt) and the evolved BoNT F(S166Y) on these constructs was measured. Many of these single mutant substrates were efficiently cleaved using the more active BoNT F(VAMP1) evolved protease. The T7-PAP(VAMP71.1) AP, carrying the VAMP1(D58G) mutation, displays lower cleavage efficiency ( FIG. 4D ) and was targeted for subsequent PACE selection. After a 72h PACE positive selection, phage with dramatically enhanced cleavage activity ( FIG. 4D ) were selected. The selected variants carried a set of highly-enriched mutations: S166Y, R240L, and Y372H. 
     Negative Selection and Evolved Proteases that Cleave VAMP7 
     Because positive selection generally results in broadened activity with low specificity, it was important to develop a negative selection strategy to select for BoNT F proteases with high specificity for the desired VAMP7 sequence. Here, a modified T7 polymerase that selectively transcribes from the T3 polymerase target sequence was fused with T7 lysozyme to generate an orthogonal protease-activated polymerase (T3neg-PAP) and was coupled to expression of a dominant negative mutant of gene III, pIII-neg, upon cleavage by non-selective BoNT (e.g., a BoNT having an undesired proteolytic activity). Competitive expression of pIII-neg effectively suppresses phage propagation by generating phage incapable of infecting new hosts, and is initiated upon cleavage of an off-target sequence in the protease-sensitive linker of T3neg ( FIG. 5 ). Because this gene cassette (containing the orthogonal polymerase and pIII-neg) are encoded on a separate plasmid, the concentration of substrate (T3neg) and the amount of pIII-neg produced per hydrolysis event can be modulated through plasmid copy number, promoter engineering, and ribosome binding sequence optimization of the resulting mRNA transcript. This allows for tunable negative selection stringency, and increases the likelihood that selective BoNT mutants can be enriched. Certain evolutionary trajectories between VAMP1 and VAMP7 cover a large number of non-natural SNARE protein sequences. Therefore, in order to obtain high specificity, negative selection against a only relatively small number of naturally-occurring off-target substrates will be performed. In some embodiments, a VAMP1-linked T3neg-PAP selects against the most likely off-target substrate for the evolved protease. Additional in vitro substrate profiling can be performed using related SNARE proteins such as VAMP2, VAMP3, and VAMP8 to ensure a high degree of specificity for the target VAMP7. 
     Characterization of Evolved BoNT Proteases 
     The chemical and biological properties evolved BoNT proteases are investigated. Recombinant expression and isolation of BoNT F LC proteases was performed and is sufficient to obtain material for in vitro cleavage assays. Single-mutant reversions of the evolved BoNT proteases are assayed to interrogate their role in controlling enzymatic activity and specificity. SNARE substrate cleavage is assayed by both gel electrophoresis and LC/MS to determine enzyme kinetics. Stability (both thermal and proteolytic) of the evolved protease is assessed in quantitative assays. The therapeutic potential of BoNT variants to bring about selective membrane fusion blockade by cleaving VAMP7 is investigated by characterizing the ability of the variants to function as targeted secretion inhibitors in human cells, for example using the MT1-MMP secretion model. Briefly, BoNT protease variant is introduced into MDA-MB-231 breast cancer cells via transfection, and extracellular matrix degradation is assayed using a fluorescent gelatin plating medium. Transwell migration assays are also performed; siRNA and anti-VAMP7 antibody data are compared with BoNT activity to determine the relative potency of BoNT protease variant treatment. Surface labeling with anti-MT1-MMP antibodies is also performed as an orthogonal assay for VAMP7 function in this invasion assay. 
     Example 2 
     PACE of BoNT Protease Variants 
     In contrast to previous selections, where protease activity and selectivity is dictated by a short (approximately 7 amino acid) peptide sequence, BoNT LC serotypes recognize an extended sequence of their cognate SNARE protein substrates. Data indicate that that a 60 amino acid fragment of VAMP1, extending from residues 28-87, serves as a suitable linker between T7 RNAP and T7 lysozyme, affording up to 3-fold activation of the polymerase upon proteolytic cleavage ( FIG. 6A ). Notably, BoNT serotypes B and F perform best on this substrate in accord with their in vivo activity, while other BoNT LC serotypes exhibit PA-RNAP activation exclusively for their cognate substrates (BoNT E on SNAP25). The VAMP1 PA-RNAP has been shown to be sufficient to carry out PACE selection, and has yielded BoNT B and BoNT F variants with increased apparent VAMP1 and VAMP2 cleavage activity ( FIG. 6B ). 
     The efficiency of PACE makes practical a stepping-stone approach in which a protein evolves recognition of a successively altered series of substrates towards a dramatically altered final substrate. Alignment of VAMP1, VAMP7, and VAMP8 shows a high degree of sequence homology, but notable deviation in activity-promoting residues for both the BoNT B and BoNT F serotypes (see  FIG. 6B , BoNT B L2F and BoNT F L3E, and  FIG. 7 ). Evolutionary pathways were designed that gradually evolve wild-type protease specificities by successive introduction of each amino acid substitution (or set of substitutions) in the VAMP7/8 target sequences into the VAMP1 PA-RNAP, followed by PACE selection on the resulting AP construct. This strategy allows a stepwise accumulation of LC mutations that result in a gradual shift in protein activity toward the desired VAMP substrates. 
     As a means for interrogating the baseline promiscuity and potential evolutionary trajectories for BoNT B and F LCs, a panel of single-residue mutants in the VAMP1 PA-RNAP in which the native VAMP1 residue has been converted into the corresponding VAMP7 or VAMP8 residue were assayed. Data indicate that BoNT F LC tolerates many of the individual VAMP7 substitutions, however three of these substitutions (VAMP1 L55A, D58G, and D65I) attenuated the protease-dependent luciferase signal. Particularly difficult substitutions such as these were targeted first, in order to enter challenging selections from wild-type activity levels. Separate PACE selection for cleavage of each these mutant substrates have yielded evolved variants with improved activity against the respective mutant substrate, indicating that the designed PA-RNAP constructs facilitate evolution of BoNT proteases ( FIG. 8 ). Importantly, the ease with which new PA-RNAP constructs can be developed, and the efficiency of PACE selection enable multiple evolutionary pathways to be interrogated in parallel, thereby increasing the probability of success. For example, each of the evolved BoNT F variants in  FIG. 8  represents a different evolutionary trajectory that can be carried forward to access new substrates with increased similarity to VAMP7. 
     Example 3 
     Evolution of BoNT F by PACE 
     First-pass PACE evolution was performed on BoNT serotypes B, D, and F. Luciferase assay data indicates that BoNT B and BoNT F can be evolved to alter protease activity, for example to increase cleavage of the native VAMP1/2 substrate ( FIG. 9 ).  FIG. 10  shows one example of an evolutionary trajectory for VAMP1 to VAMP7 cleaving proteases and examples of accessory plasmids for achieving the same. 
     VAMP7 participates in phagocytosis, mitosis, cell migration, membrane repair and growth.  FIG. 11  depicts an alignment of BoNT F and BoNT B VAMP2 (a natural substrate) cleavage domains with VAMP7.  FIG. 12  provides data indicating that evolution of BoNT B and F by PACE (e.g., using AP&#39;s 977, 983 and 986 for BoNT F) resulted in BoNT variants with improved activity.  FIG. 13  shows representative data relating to validation of BoNT Light Chain (LC) selection; data indicate that evolution of BoNT F protease on VAMP1 enriches for the S166Y mutation, which confers broadly increased activity. Evolution of BoNT F (S166Y) on AP-977 also enriches strongly for mutations at position 240, which directly contacts the altered substrate residue (D-&gt;G). 
       FIG. 14  shows one example of a stepping-stone evolutionary pathway for production of BoNT F variants that cleave VAMP7.  FIG. 15  shows protease activity assays for BoNT F variants from three different experiments (Lagoons 1-6). Each experiment produced a different single mutant variant (L55A, D58G, D65I). Lagoons 1-4 were carried forward for PACE experiments using a double mutant substrate (AP-015: L55A/D58G) AP. 
       FIG. 16  shows a schematic depiction of double mutant PACE experiments to evolve BoNT F; the amino acid sequence of the double mutant VAMP1 (LSSA/D58G) is also shown. Protease-dependent luciferase assay data indicate that BoNT F variants that cleave the double mutant VAMP1 substrate were produced by PACE ( FIG. 17 ). 
       FIG. 18  shows one example of an evolutionary “stepping stone” strategy for mutation of BoNT F to cleave VAMP7.  FIG. 19  shows representative data for triple mutant (L55A/D58G/Q59E) selection of BoNT F variants. Data indicate that variants L132-L1C and L132-L3A cleave VAMP1 containing three VAMP7 mutations (L55A/D58G/Q59E). 
       FIG. 20  shows representative data for tetramutant (L55A/D58G/Q59E/K60R) selection of BoNT F variants. It was observed that several selected BoNT F variants (e.g., 216-L1C, 216-L1E, 216-L3B, 216-L3G) cleave the VAMP1 containing four VAMP7 mutations (L55A/D58G/Q59E/K60R), indicating that four of the five least permissive mutation sites in VAMP1 have been addressed.  FIG. 21  shows that activity of proteases on V44K (shown as V43 in the figure) and Q32M (shown as Q33 in the figure) can be readily evolved. BoNT F proteases tolerant of VAMP7 termini have also been observed. 
     Iterative selection on progressively more complex VAMP substrates afforded several BoNT F variants that cleave VAMP7 ( FIG. 22 ).  FIG. 23  shows an alignment of VAMP1 and VAMP7 amino acid sequences, along with AP-V7-194KL, which contains seven VAMP7 mutations (V44K/K53L/L55A/D58G/Q59E/K60R/D65I). Table 3 below shows mutations observed in several BoNT F variants after PACE with AP-V7-194KL for 48 h, followed by AP-V7-194KL+AP-092 for 24 hours, followed by AP-092 for 48 hours. 
     
       
         
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
                 TABLE 3 
               
               
                   
               
             
            
               
                 L1 
                 a 
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
                   
                   
               
               
                   
                 b 
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
                 Y210H 
               
               
                   
                 c 
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
               
               
                   
                 d 
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
               
               
                   
                 e 
                   
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
                   
                 T214I 
               
               
                   
                 f 
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
               
               
                   
                 g 
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
               
               
                   
                 h 
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
               
               
                 L2 
                 a 
                   
                   
                   
                 V106A 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                 E200G 
               
               
                   
                 b 
                   
                   
                   
                 V106A 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                 E200G 
               
               
                   
                 c 
                   
                   
                   
                 V106A 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                 E200G 
               
               
                   
                 d 
                   
                   
                 N76D 
                 V106A 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                 E200G 
               
               
                   
                 e 
                   
                   
                   
                 V106A 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                 E200G 
               
               
                   
                 f 
                   
                   
                   
                 V106A 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                 E200G 
               
               
                   
                 g 
                   
                   
                   
                 V106A 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                 E200G 
               
               
                   
                 h 
                   
                   
                   
                 V106A 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                 E200G 
               
               
                 L3 
                 a 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
               
               
                   
                 b 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
               
               
                   
                 c 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
               
               
                   
                 d 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                 Y199H 
                 E200G 
               
               
                   
                 e 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
               
               
                   
                 f 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
               
               
                   
                 g 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
               
               
                   
                 h 
                   
                 S70F 
                   
                   
                 E164K 
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 L1 
                 a 
                   
                 S224I 
                 R240F 
                   
                   
                   
                 R303H 
                 P309T 
                   
                   
                 F360L 
               
               
                   
                 b 
                   
                 S224I 
                 R240F 
                   
                   
                   
                   
                   
                   
                   
                 F360L 
               
               
                   
                 c 
                   
                 S224I 
                 R240F 
                   
                   
                   
                   
                   
                   
                   
                 F360L 
               
               
                   
                 d 
                 E215G 
                   
                 R240L 
                   
                   
                   
                   
                   
                   
                   
                 F360L 
               
               
                   
                 e 
                   
                 S224I 
                 R240F 
                   
                   
                   
                   
                   
                   
                   
                 F360L 
               
               
                   
                 f 
                   
                 S224I 
                 R240F 
                   
                   
                   
                   
                   
                   
                   
                 F360L 
               
               
                   
                 g 
                   
                 S224I 
                 R240F 
                   
                   
                   
                   
                   
                   
                   
                 F360L 
               
               
                   
                 h 
                 E215G 
                   
                 R240L 
                 Y244C 
                   
                   
                   
                   
                   
                   
                 F360L 
               
               
                 L2 
                 a 
                   
                 S224I 
                 R240L 
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                   
                 b 
                   
                 S224I 
                 R240L 
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                   
                 c 
                   
                 S224I 
                 R240L 
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                   
                 d 
                   
                 S224I 
                 R240L 
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                   
                 e 
                   
                 S224I 
                 R240L 
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                   
                 f 
                   
                 S224I 
                 R240L 
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                   
                 g 
                   
                 S224I 
                 R240L 
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                   
                 h 
                   
                 S224I 
                 R240L 
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 L3 
                 a 
                   
                 S224I 
                 R240F 
                   
                   
                   
                   
                   
                 T335S 
                   
                 F360L 
               
               
                   
                 b 
                   
                 S224I 
                 R240F 
                   
                   
                 N276T 
                   
                   
                 T335S 
                   
                 F360L 
               
               
                   
                 c 
                   
                 S224I 
                 R240F 
                   
                   
                   
                   
                   
                 T335S 
                   
                 F360L 
               
               
                   
                 d 
                   
                 S224I 
                 R240F 
                   
                   
                   
                   
                   
                 T335S 
                   
                 F360L 
               
               
                   
                 e 
                   
                 S224I 
                 R240F 
                   
                   
                   
                   
                   
                 T335S 
                   
                 F360L 
               
               
                   
                 f 
                   
                 S224I 
                 R240F 
                   
                 A258S 
                 N276S 
                   
                   
                   
                   
                 F360L 
               
               
                   
                 g 
                   
                 S224I 
                 R240F 
                   
                   
                   
                   
                   
                 T335S 
                   
                 F360L 
               
               
                   
                 h 
                   
                 S224I 
                 R240F 
                   
                   
                   
                   
                   
                   
                   
                 F360L 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                 L1 
                 a 
                   
                 Y372H 
                   
                   
                 P410L 
                   
                 420(AWLRKS*) 
                   
               
               
                   
                   
                 b 
                   
                 Y372H 
                   
                   
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                   
                 c 
                   
                 Y372H 
                   
                   
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                   
                 d 
                 K371E 
                 Y372H 
                   
                   
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                   
                 e 
                   
                 Y372H 
                   
                   
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                   
                 f 
                   
                 Y372H 
                   
                   
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                   
                 g 
                   
                 Y372H 
                   
                   
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                   
                 h 
                 K371E 
                 Y372H 
                   
                   
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                 L2 
                 a 
                   
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                   
                 b 
                   
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                   
                 c 
                   
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                   
                 d 
                   
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                   
                 e 
                   
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                   
                 f 
                   
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                   
                 g 
                   
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                   
                 h 
                   
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                 L3 
                 a 
                   
                 Y372H 
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                 E423K 
               
               
                   
                   
                 b 
                   
                 Y372H 
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                 E423K 
               
               
                   
                   
                 c 
                   
                 Y372H 
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                 E423K 
               
               
                   
                   
                 d 
                   
                 Y372H 
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                 E423K 
               
               
                   
                   
                 e 
                   
                 Y372H 
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                 E423K 
               
               
                   
                   
                 f 
                   
                 Y372H 
                 L375R 
                 N396H 
                 P410L 
                   
                   
                 E423K 
               
               
                   
                   
                 g 
                   
                 Y372H 
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                 E423K 
               
               
                   
                   
                 h 
                   
                 Y372H 
                 L375R 
                 N396H 
                 P410L 
                   
                   
                 E423K 
               
               
                   
                   
               
            
           
         
       
     
     Several VAMP7-cleaving BoNT F variants were expressed in vitro.  FIG. 24  shows protein blot analysis for protein expression of two BoNT F evolved variants (2020 L2A, 2020 L3A). 
     VAMP7-evolved proteases were then used to screen a VAMP8 double mutant substrate panel.  FIG. 25  shows a schematic diagram of an alignment of VAMP1 and VAMP8 amino acid sequences and double mutant accessory plasmids (APs) used in the screen. Data indicates that VAMP7-evolved BoNT F proteases have a broadened activity profile ( FIG. 26 ). 
       FIG. 27  shows an alignment of VAMP1 and VAMP8 amino acid sequences, along with several APs used to evolve VAMP8-cleaving BoNT F variants. Data indicate that VAMP8 APs have high background but BoNT F variants that cleave VAMP8 were identified ( FIG. 27 ). 
     Example 4 
     Evolution of BoNT E by PACE 
     Wild-type BoNT E cleaves SNAP25 protein ( FIG. 28 ). This example describes evolution of BoNT E to cleave the non-native substrates, such as SNAP23 and PTEN proteins. First, cleavage of SNAP25 residues 166-186 was examined by a protease-dependent luminescence assay. Data indicate that mutation of the residue at position 179 (e.g., D179K) of SNAP25 abolished protease activity by BoNT E ( FIG. 29 ). 
     Two different PACE experiments were performed. In the first experiment (Lagoons 1 and 2), BoNT E protein variants were evolved in the presence of a two-substrate gradient (SNAP25 and SNAP25 D179K). In the second experiment (Lagoons 3 and 4), BoNT E protein variants were evolved in the presence of a different two-substrate gradient (SNAP25 and SNAP23). Table 4 below shows mutations of variants produced in each experiment. Data indicate that several BoNT E variants that cleave SNAP 23 were evolved ( FIG. 30 ) 
     
       
         
           
               
               
               
               
               
               
               
               
               
               
               
               
             
               
                 TABLE 4 
               
               
                   
               
             
            
               
                 L1 
                 A 
                 I18V 
                   
                   
                 L89P 
                   
                   
                   
                   
                   
                 K225E 
               
               
                   
                 B 
                 I18V 
                   
                   
                 L89P 
                   
                   
                   
                   
                   
                 K225E 
               
               
                   
                 C 
                 I18V 
                   
                   
                   
                   
                 E154G 
                   
                   
                 I199T 
                 K225E 
               
               
                   
                 D 
                 I18V 
                   
                   
                 L89P 
                   
                   
                   
                   
                   
                 K225E 
               
               
                   
                 E 
                   
                   
                   
                   
                   
                 E154G 
                   
                   
                   
                 K225E 
               
               
                   
                 F 
                 I18V 
                   
                   
                   
                   
                 E154G 
                   
                   
                   
                 K225E 
               
               
                   
                 G 
                   
                 Q27H 
                   
                   
                   
                   
                   
                   
                   
                 K225E 
               
               
                   
                 H 
                   
                 Q27H 
                   
                   
                   
                   
                   
                   
                   
                 K225E 
               
               
                 L2 
                 A 
                   
                   
                 E28K 
                   
                   
                   
                   
                   
                   
                 K225E 
               
               
                   
                 B 
                   
                   
                 E28K 
                   
                   
                   
                   
                   
                   
                 K225E 
               
               
                   
                 C 
                   
                   
                 E28K 
                   
                   
                   
                   
                   
                   
                 K225E 
               
               
                   
                 D 
                   
                   
                 E28K 
                   
                   
                   
                   
                   
                   
                 K225E 
               
               
                   
                 E 
                   
                   
                 E28K 
                   
                   
                   
                   
                   
                   
                 K225E 
               
               
                   
                 F 
                   
                   
                 E28K 
                   
                   
                   
                   
                   
                   
                 K225E 
               
               
                 L3 
                 A 
                 I18V 
                   
                   
                   
                   
                 E154G 
                 R168K 
                   
                   
                 K225E 
               
               
                   
                 B 
                 I18V 
                   
                   
                   
                   
                 E154G 
                   
                   
                   
                 K225E 
               
               
                   
                 C 
                   
                   
                   
                   
                   
                 E154G 
                   
                   
                   
                 K225E 
               
               
                   
                 D 
                 I18V 
                   
                   
                   
                   
                 E154G 
                 R168K 
                   
                   
                 K225E 
               
               
                   
                 E 
                   
                   
                   
                   
                   
                 E154G 
                   
                 S187F 
                   
                 K225E 
               
               
                   
                 F 
                 I18V 
                   
                   
                   
                   
                 E154G 
                 R168K 
                   
                   
                 K225E 
               
               
                   
                 H 
                 I18V 
                   
                   
                   
                   
                 E154G 
                 R168K 
                   
                   
                 K225E 
               
               
                 L4 
                 A 
                   
                   
                 E28K 
                   
                 Q141K 
                 E154G 
                   
                   
                   
                 K225E 
               
               
                   
                 B 
                   
                   
                 E28K 
                   
                 Q141K 
                 E154G 
                   
                   
                   
                 K225E 
               
               
                   
                 C 
                   
                   
                 E28K 
                   
                 Q141K 
                 E154G 
                   
                   
                   
                 K225E 
               
               
                   
                 E 
                   
                   
                 E28K 
                   
                 Q141K 
                 E154G 
                   
                   
                   
                 K225E 
               
               
                   
                 F 
                   
                   
                 E28K 
                   
                 Q141K 
                 E154G 
                   
                   
                   
                 K225E 
               
               
                   
                 G 
                   
                   
                 E28K 
                   
                 Q141K 
                 E154G 
                   
                   
                   
                 K225E 
               
               
                   
                 H 
                   
                   
                 E28K 
                   
                 Q141K 
                 E154G 
                   
                   
                   
                 K225E 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                 L1 
                 A 
                   
                   
                   
                   
                 V265G 
                   
                   
                   
                   
               
               
                   
                   
                 B 
                   
                   
                   
                   
                   
                   
                   
                   
                 L404* 
               
               
                   
                   
                 C 
                   
                   
                 N258S 
               
               
                   
                   
                 D 
               
               
                   
                   
                 E 
               
               
                   
                   
                 F 
               
               
                   
                   
                 G 
                   
                   
                   
                   
                   
                   
                   
                   
                 L404* 
               
               
                   
                   
                 H 
                   
                   
                   
                   
                   
                   
                   
                   
                 L404* 
               
               
                   
                 L2 
                 A 
                 C231R 
                   
                   
                   
                   
                   
                   
                   
                 L404* 
               
               
                   
                   
                 B 
                 C231R 
                   
                   
                   
                   
                   
                   
                   
                 L404* 
               
               
                   
                   
                 C 
                 C231R 
                   
                   
                   
                   
                   
                   
                   
                 L404* 
               
               
                   
                   
                 D 
                 C231R 
               
               
                   
                   
                 E 
                 C231R 
                   
                   
                 N261D 
               
               
                   
                   
                 F 
                 C231R 
                   
                   
                   
                   
                   
                   
                   
                 L404* 
               
               
                   
                 L3 
                 A 
                   
                   
                   
                   
                   
                   
                   
                 P398L 
               
               
                   
                   
                 B 
               
               
                   
                   
                 C 
                   
                   
                   
                   
                   
                 D270N 
               
               
                   
                   
                 D 
                   
                   
                   
                   
                   
                   
                   
                 P398L 
               
               
                   
                   
                 E 
               
               
                   
                   
                 F 
                   
                   
                   
                   
                   
                   
                   
                 P398L 
               
               
                   
                   
                 H 
                   
                   
                   
                   
                   
                   
                   
                 P398L 
               
               
                   
                 L4 
                 A 
                 C231R 
                 I233T 
                   
                   
                   
                   
                 I396S 
               
               
                   
                   
                 B 
                 C231R 
                 I233T 
                   
                   
                   
                   
                 I396S 
               
               
                   
                   
                 C 
                 C231R 
                 I233T 
                   
                   
                   
                   
                 I396S 
               
               
                   
                   
                 E 
                 C231R 
                 I233T 
                   
                   
                   
                   
                 I396S 
               
               
                   
                   
                 F 
                 C231R 
                 I233T 
                   
                   
                   
                   
                 I396S 
               
               
                   
                   
                 G 
                 C231R 
                 I233T 
                   
                   
                   
                   
                 I396S 
               
               
                   
                   
                 H 
                 C231R 
                 I233T 
                   
                   
                   
                   
                 I396S 
               
               
                   
                   
               
            
           
         
       
     
     Next, whether BoNT E variants that cleave a therapeutic target could be evolved was investigated.  FIG. 31  shows a portion of the SNAP25 amino acid sequence and the peptide bonds at which wild-type BoNT A and BoNT E proteases that cleave SNAP25, but not SNAP23.  FIG. 32  shows a stepping stone schematic for PACE of BoNT E to cleave the therapeutic target phosphatase and tensin homolog (PTEN). Tables 5 and 6 below shows mutations of BoNT E variants produced in each experiment. 
     
       
         
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
                 TABLE 5 
               
               
                   
               
             
            
               
                 L1A 
                   
                   
                 K22R 
                   
                 E28K 
                   
                   
                   
                   
                 E154G 
                   
                   
                   
               
               
                 L1B 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 E154G 
                   
                 T160A 
               
               
                 L1C 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 E154G 
                   
                 T160A 
               
               
                 L1D 
                   
                   
                 K22R 
                   
                 E28K 
                   
                   
                   
                   
                 E154G 
               
               
                 L1E 
                   
                   
                 K22R 
                   
                 E28K 
                   
                   
                   
                   
                 E154G 
               
               
                 L1F 
                   
                   
                 K22R 
                   
                 E28K 
                   
                   
                   
                   
                 E154G 
               
               
                 L1G 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 E154G 
                   
                 T160A 
               
               
                 L1H 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 E154G 
               
               
                 L2A 
                   
                   
                   
                   
                 E28K 
                   
                   
                   
                   
                   
                 E159A 
                 T160A 
                 N161H 
               
               
                 L2B 
                   
                   
                   
                   
                 E28K 
                   
                   
                   
                 D128G 
                   
                 E159A 
                 T160A 
                 N161H 
               
               
                 L2C 
                   
                 I21M 
                   
                   
                 E28K 
                   
                   
                   
                   
                   
                 E159A 
                 T160A 
                 N161H 
               
               
                 L2D 
                   
                 I21M 
                   
                   
                 E28K 
                   
                   
                   
                   
                   
                 E159A 
                 T160A 
                 N161H 
               
               
                 L2E 
                   
                   
                   
                   
                 E28K 
                   
                   
                   
                   
                   
                 E159A 
                 T160A 
                 N161H 
               
               
                 L2F 
                   
                   
                   
                 Q27H 
                   
                 S99A 
                 G101S 
                   
                   
                   
                 E159L 
                   
                 N161Y 
               
               
                 L2H 
                   
                   
                   
                   
                 E28K 
                   
                   
                 T119K 
                   
                   
                 E159A 
                 T160A 
                 N161H 
               
               
                 L3A 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 E159C 
                   
                 N161W 
               
               
                 L3B 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 E159C 
                 T160S 
                 N161W 
               
               
                 L3C 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 E159C 
                 T160S 
                 N161W 
               
               
                 L3D 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 E159C 
                   
                 N161W 
               
               
                 L3E 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 E159C 
                 T160S 
                 N161W 
               
               
                 L3F 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 E159C 
                 T160S 
                 N161W 
               
               
                 L3G 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 E159C 
                   
                 N161W 
               
               
                 L3H 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 E159C 
                 T160S 
                 N161W 
               
               
                 L4A 
                   
                   
                   
                   
                 E28K 
               
               
                 L4B 
                   
                   
                   
                   
                 E28K 
               
               
                 L4C 
                   
                   
                   
                   
                 E28K 
               
               
                 L4D 
                   
                   
                   
                   
                 E28K 
               
               
                 L4E 
                   
                   
                   
                   
                 E28K 
               
               
                 L4F 
                   
                   
                   
                   
                 E28K 
               
               
                 L4G 
                 S6G 
                   
                   
                   
                 E28K 
               
               
                 L4H 
                   
                   
                   
                   
                 E28K 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 L1A 
                   
                 S163R 
                   
                   
                   
                   
                 K225H 
                   
                   
                   
               
               
                 L1B 
                   
                   
                   
                   
                   
                   
                 K225H 
                 C231R 
               
               
                 L1C 
                   
                   
                   
                   
                   
                   
                 K225H 
                 C231R 
               
               
                 L1D 
                   
                 S163R 
                   
                   
                   
                   
                 K225H 
               
               
                 L1E 
                   
                 S163R 
                   
                   
                   
                   
                 K225H 
               
               
                 L1F 
                   
                 S163R 
                   
                   
                   
                   
                 K225H 
               
               
                 L1G 
                   
                   
                   
                   
                   
                   
                 K225H 
                 C231R 
               
               
                 L1H 
                 S162A 
                   
                 M172R 
                   
                   
                   
                 K225H 
               
               
                 L2A 
                   
                   
                   
                 S174A 
               
               
                 L2B 
                   
                   
                   
                 S174A 
               
               
                 L2C 
                   
                   
                   
                 S174A 
               
               
                 L2D 
                   
                   
                   
                 S174A 
               
               
                 L2E 
                   
                 S163R 
                   
                 S174A 
                   
                   
                   
                   
                   
                 K245R 
               
               
                 L2F 
                 S162Q 
                   
                   
                   
                   
                   
                   
                   
                 I232T 
               
               
                 L2H 
                   
                   
                   
                 S174A 
               
               
                 L3A 
                   
                   
                   
                   
                   
                   
                 K225L 
                 C231R 
               
               
                 L3B 
                   
                   
                   
                   
                 I199M 
                   
                 K225L 
                 C231R 
               
               
                 L3C 
                   
                   
                   
                   
                   
                   
                 K225L 
                 C231R 
               
               
                 L3D 
                   
                   
                   
                   
                   
                 A224S 
                 K225L 
                 C231R 
                   
                 K245R 
               
               
                 L3E 
                   
                   
                   
                   
                 I199M 
                   
                 K225L 
                 C231R 
               
               
                 L3F 
                   
                   
                   
                   
                   
                   
                 K225L 
                 C231R 
               
               
                 L3G 
                   
                   
                   
                   
                   
                   
                 K225L 
                 C231R 
               
               
                 L3H 
                   
                   
                   
                   
                 I199M 
                   
                 K225L 
                 C231R 
               
               
                 L4A 
                   
                   
                   
                   
                   
                   
                 K225L 
                   
                 I232T 
               
               
                 L4B 
                   
                   
                   
                   
                   
                   
                 K225L 
                   
                 I232T 
               
               
                 L4C 
                   
                   
                   
                   
                   
                   
                 K225L 
                   
                 I232T 
               
               
                 L4D 
                   
                   
                   
                   
                   
                   
                 K225L 
                   
                 I232T 
               
               
                 L4E 
                   
                   
                   
                   
                   
                   
                 K225L 
                   
                 I232T 
               
               
                 L4F 
                   
                   
                   
                   
                   
                   
                 K225L 
                   
                 I232T 
               
               
                 L4G 
                   
                   
                   
                   
                   
                   
                 K225L 
                   
                 I232T 
               
               
                 L4H 
                   
                   
                   
                   
                   
                   
                 K225L 
                   
                 I232T 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                 L1A 
                   
                   
                   
                   
                 I352A 
                   
                   
                 Y357C 
                   
                   
               
               
                   
                 L1B 
                   
                 I263V 
                   
                   
                   
                   
                   
                 Y357C 
               
               
                   
                 L1C 
                   
                 I263V 
                   
                   
                   
                   
                   
                 Y357C 
               
               
                   
                 L1D 
                   
                   
                   
                   
                 I352A 
                   
                   
                 Y357C 
               
               
                   
                 L1E 
                   
                   
                   
                   
                 I352A 
                   
                   
                 Y357C 
               
               
                   
                 L1F 
                   
                   
                   
                   
                 I352A 
                   
                   
                 Y357W 
               
               
                   
                 L1G 
                   
                 I263V 
                   
                   
                   
                   
                   
                 Y357C 
                 I399S 
               
               
                   
                 L1H 
               
               
                   
                 L2A 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
                   
                   
                 L404* 
               
               
                   
                 L2B 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
                   
                   
                 L404* 
               
               
                   
                 L2C 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
                   
                   
                 L404* 
               
               
                   
                 L2D 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
                   
                   
                 L404* 
               
               
                   
                 L2E 
                   
                   
                 V265G 
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 L2F 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 L2H 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
                   
                   
                 L404* 
               
               
                   
                 L3A 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 L3B 
                   
                   
                 V265G 
                   
                   
                 Q354R 
                   
                 Y357C 
               
               
                   
                 L3C 
                   
                   
                   
                   
                   
                   
                   
                 Y357C 
               
               
                   
                 L3D 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 L3E 
                   
                   
                 V265G 
                 A313S 
                   
                 Q354R 
                   
                 Y357C 
               
               
                   
                 L3F 
                   
                   
                   
                   
                   
                   
                   
                 Y357C 
                   
                 L404* 
               
               
                   
                 L3G 
                 I247V 
                   
                 V265G 
                   
                 I352V 
                   
                   
                 Y357C 
               
               
                   
                 L3H 
                   
                   
                 V265G 
                   
                   
                   
                   
                 Y357C 
               
               
                   
                 L4A 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 L4B 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 L4C 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 L4D 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 L4E 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 L4F 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 L4G 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 L4H 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
               
               
               
               
               
               
               
               
               
               
             
               
                 TABLE 6 
               
               
                   
               
             
            
               
                 L2A 
                 Q27H 
                 I35L 
                   
                 S99A 
                 G101S 
                   
                 E159L 
                 N161Y 
                 S162Q 
                 M172R 
               
               
                 L2B 
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                   
                 E159L 
                 N161Y 
                 S162Q 
                 M172K 
               
               
                 L2C 
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                   
                 E159L 
                 N161Y 
                 S162Q 
                 M172K 
               
               
                 L2D 
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 V132G 
                 E159L 
                 N161Y 
                 S162Q 
                 M172K 
               
               
                 L2F 
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                   
                 E159L 
                 N161Y 
                 S162Q 
                 M172K 
               
               
                 L2G 
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                   
                 E159L 
                 N161Y 
                 S162Q 
                 M172K 
               
               
                 L2H 
                 Q27H 
                   
                 D53Y 
                 S99A 
                 G101S 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                   
                 L2A 
                 I199V 
                 I232T 
                 N243S 
                 T273I 
                   
                 Q354R 
                 Y355H 
                 L404* 
               
               
                   
                 L2B 
                   
                 I232T 
                   
                   
                 I302M 
                 Q354R 
                 Y355H 
               
               
                   
                 L2C 
                   
                 I232T 
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 L2D 
                   
                 I232T 
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 L2F 
                   
                 I232T 
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 L2G 
                   
                 I232T 
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 L2H 
                   
                 I232T 
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
               
            
           
         
       
     
     Simultaneous positive (proB stringency) and negative selection (SNAP25, mixing from proA to proB) of BoNT E variants that cleave PTEN was performed. Evolution of several BoNT E variants that cleave PTEN was observed ( FIG. 33 ). Table 7 below shows mutations of BoNT E variants produced in this experiment. 
     
       
         
           
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
                 TABLE 7 
               
               
                   
               
             
            
               
                 L2 
                 A 
                   
                 Q27H 
                 F29L 
                   
                 S99A 
                 G101S 
                 N118D 
                 G127S 
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 L2 
                 B 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 L2 
                 C 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 L2 
                 D 
                 C26Y 
                 Q27H 
                   
                 Y68H 
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 L2 
                 E 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 L2 
                 F 
                   
                 Q27H 
                   
                   
                 S99T 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 L2 
                 G 
                 C26Y 
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 L2 
                 H 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 L2 
                 I 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 L2 
                 J 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 L2 
                 K 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 L2 
                 L 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 L2 
                 M 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 L2 
                 N 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 L2 
                 O 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 L2 
                 P 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                 L2 
                 A 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
                   
                   
               
               
                   
                 L2 
                 B 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 L2 
                 C 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 L2 
                 D 
                 S163R 
                 M172K 
                 I232T 
                 N238S 
                   
                 Q354R 
                 Y357H 
               
               
                   
                 L2 
                 E 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
                   
                 I409T 
               
               
                   
                 L2 
                 F 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
                 L404* 
               
               
                   
                 L2 
                 G 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357H 
               
               
                   
                 L2 
                 H 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 L2 
                 I 
                 S163R 
                 M172K 
                 I232T 
                   
                 Q295R 
                 Q354R 
                 Y357P 
               
               
                   
                 L2 
                 J 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357H 
               
               
                   
                 L2 
                 K 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 L2 
                 L 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 L2 
                 M 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 L2 
                 N 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 L2 
                 O 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 L2 
                 P 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                   
               
            
           
         
       
     
     An evolved BoNT E variant (L2F 031017) was expressed and purified by increasing concentrations of imidazole ( FIG. 34 ). A proteolysis assay was performed, using SNAP25 and PTEN as substrates. Data indicate that BoNT E L2F cleaves both SNAP25 and PTEN ( FIG. 35 ). PACE using negative selection results in improved PTEN cleavage, for example by L2B variant as shown in  FIG. 36 . 
     PACE Evolution Tables 
       
     
       
         
           
               
             
               
                 TABLE 8 
               
               
                   
               
               
                 BoNT E PACE 1 Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 1 
                   
                 Q27K 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 2 
                   
                 Q27K 
               
               
                 3 
                   
                   
                 E28K 
               
               
                 4 
               
               
                 5 
                   
                 Q27K 
               
               
                 6 
                   
                   
                 E28K 
               
               
                 7 
                   
                   
                 E28K 
               
               
                 8 
               
               
                 9 
                   
                   
                   
                   
                   
                   
                   
                   
                 E159A 
                 N161H 
               
               
                 10 
                   
                 Q27K 
                   
                   
                   
                   
                   
                   
                 E159A 
                 N161H 
               
               
                 11 
                   
                 Q27K 
               
               
                 12 
                   
                   
                   
                   
                   
                   
                   
                   
                 E159A 
               
               
                 13 
                   
                   
                   
                   
                   
                   
                   
                   
                 E159A 
               
               
                 14 
                   
                   
                   
                   
                   
                   
                   
                   
                 E159L 
                 N161Y 
               
               
                 15 
                   
                   
                 E28K 
                   
                   
                   
                   
                   
                 E159S 
                 N161H 
               
               
                 16 
                   
                   
                   
                   
                   
                   
                   
                   
                 E159A 
                 N161Y 
               
               
                 17 
                 Y20C 
                   
                 E28K 
                 E78G 
                   
                   
                 D128A 
                 A129Q 
               
               
                 18 
                   
                   
                   
                   
                   
                   
                 D128G 
                 A129G 
               
               
                 19 
                   
                   
                   
                   
                   
                   
                 D128G 
                 A129Q 
               
               
                 20 
                   
                   
                   
                   
                   
                   
                 D128A 
                 A129Q 
               
               
                 21 
                   
                   
                   
                   
                   
                   
                 D128A 
                 A129Q 
               
               
                 22 
               
               
                 23 
               
               
                 24 
                   
                   
                   
                   
                   
                   
                 D128G 
                 A129Q 
               
               
                 25 
               
               
                 26 
                   
                   
                 E28K 
               
               
                 27 
                   
                   
                 E28K 
               
               
                 28 
                   
                   
                   
                   
                   
                 G101S 
               
               
                 29 
                   
                   
                 E28K 
                   
                 L98P 
               
               
                 30 
               
               
                 31 
                   
                   
                 E28K 
               
               
                 32 
                   
                   
                   
                   
                   
                 G101S 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 K225L 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                 K225H 
                   
                 I232S 
               
               
                   
                   
                 K225L 
                   
                   
                   
                 I352T 
               
               
                   
                   
                 K225L 
               
               
                   
                   
                 K225H 
               
               
                   
                   
                 K225H 
                   
                 I232T 
               
               
                   
                   
                 K225H 
               
               
                   
                   
                 K225L 
               
               
                   
                   
                   
                   
                 I232S 
               
               
                   
                   
                   
                   
                 I232S 
               
               
                   
                   
                   
                   
                 I232T 
               
               
                   
                   
                   
                   
                 I232T 
               
               
                   
                   
                   
                   
                   
                 V345I 
                   
                   
                   
                   
                 L404* 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 I399S 
               
               
                   
                   
                   
                 I227T 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 T400A 
               
               
                   
                   
                   
                   
                 I232T 
               
               
                 I165V 
                   
                   
                   
                 I232T 
               
               
                   
                 R168K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F358L 
               
               
                   
                   
                   
                   
                 I232S 
                   
                   
                   
                   
                 T400K 
               
               
                   
                 R168K 
                   
                   
                 I232T 
               
               
                   
               
               
                 Unique amino acid sequences of BoNT E PACE 1 variants are provided in SEQ ID NOs: 1-26. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 9 
               
               
                   
               
               
                 BoNT E PACE 2 Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 1 
                   
                   
                   
                   
                 L98P 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 2 
                   
                   
                 E28K 
               
               
                 3 
                   
                 Q27R 
                   
                   
                 L98P 
                   
                   
                   
                   
                   
                 N197K 
                   
                 I199M 
               
               
                 4 
                   
                   
                 E28K 
               
               
                 5 
                   
                   
                   
                   
                   
                   
                   
                 Y171C 
                   
                   
                   
                   
                   
                 A224S 
               
               
                 6 
                 K22S 
                   
                 E28K 
                   
                   
                   
                   
                   
                   
                 E184G 
               
               
                 7 
               
               
                 8 
               
               
                 9 
               
               
                 10 
                   
                   
                   
                 V47I 
                 L98P 
               
               
                 11 
               
               
                 12 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 N197K 
               
               
                 13 
                   
                   
                 E28K 
                   
                   
                 E159S 
                 N161H 
                   
                 M172S 
               
               
                 14 
                   
                   
                   
                   
                   
                 E159S 
                 N161W 
               
               
                 15 
                   
                 Q27R 
                   
                   
                 L98P 
                 E159W 
                 N161W 
               
               
                 16 
                   
                   
                   
                   
                   
                 E159Q 
                 N161Y 
               
               
                 17 
                   
                   
                   
                   
                   
                 E159A 
                 N161W 
                   
                 M172V 
               
               
                 18 
                   
                 Q27R 
                   
                   
                 L98P 
                 E159A 
                 N161V 
               
               
                 19 
                   
                   
                   
                   
                 L98P 
                 E159C 
                 N161W 
               
               
                 20 
                   
                   
                 E28K 
               
               
                 21 
               
               
                 22 
                   
                   
                 E28K 
               
               
                 23 
                   
                   
                   
                   
                   
                 E159A 
                 N161H 
               
               
                 24 
                   
                   
                 E28K 
                   
                   
                   
                   
                   
                   
                   
                   
                 S198G 
               
               
                 25 
               
               
                 26 
                   
                   
                   
                   
                   
                 E159A 
                 N161H 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 K225H 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 K225L 
               
               
                 K225H 
               
               
                 K225L 
                 I227V 
                   
                   
                   
                   
                   
                 I352T 
                   
                   
                   
                   
                   
                 L404* 
               
               
                 K225H 
               
               
                 K225H 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 Y357C 
               
               
                   
                   
                   
                 I232T 
               
               
                   
                   
                   
                 I232T 
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
                   
                 I232T 
                   
                 V265G 
               
               
                   
                   
                   
                 I232T 
                   
                   
                   
                   
                   
                   
                   
                 S372G 
               
               
                   
                   
                   
                 I232T 
                   
                   
                   
                   
                 Q354R 
                 Y355H 
                   
                   
                 N379K 
               
               
                   
                   
                   
                 I232T 
                   
                   
                 S314A 
                   
                 Q354R 
                 Y355H 
               
               
                 K225H 
                   
                 C231G 
               
               
                 K225L 
                   
                   
                   
                   
                   
                   
                 I352T 
                   
                   
                   
                   
                   
                   
                 I409L 
               
               
                 K225H 
               
               
                 K225H 
               
               
                 K225L 
                   
                 C231R 
               
               
                 K225H 
               
               
                 K225L 
                   
                 C231R 
               
               
                 K225L 
                   
                   
                 I232T 
               
               
                 K225L 
                   
                   
                 I232T 
               
               
                 K225L 
                   
                   
                 I232T 
               
               
                 K225Y 
                   
                   
                 I232T 
                 N242S 
               
               
                 K225L 
                   
                   
                 I232T 
               
               
                 K225L 
                   
                   
                 I232T 
               
               
                 K225H 
               
               
                   
               
               
                 Unique amino acid sequences of BoNT E PACE 2 variants are provided in SEQ ID NOs: 27-49. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 10 
               
               
                   
               
               
                 BoNT E PACE 3 Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 1 
                   
                   
                   
                 E148G 
                   
                 T160A 
                   
                 Y171C 
                   
                   
                   
                   
                   
               
               
                 2 
                   
                   
                   
                 E148G 
               
               
                 3 
                   
                   
                   
                 E148G 
               
               
                 4 
                   
                   
                   
                 E148G 
                   
                   
                   
                   
                   
                   
                 F186L 
               
               
                 5 
                   
                   
                   
                 E148G 
                   
                   
                   
                 Y171C 
               
               
                 6 
                   
                   
                 Q141R 
                   
                 E159A 
                 T160A 
                 N161H 
                   
                   
                 S174A 
               
               
                 7 
                   
                   
                   
                   
                 E159A 
                 T160A 
                 N161H 
                   
                   
                 S174A 
               
               
                 8 
                   
                   
                   
                   
                 E159A 
                 T160A 
                 N161H 
                   
                   
                 S174A 
               
               
                 9 
                   
                   
                   
                   
                 E159A 
                 T160A 
                 N161H 
                   
                   
                 S174A 
               
               
                 10 
                   
                   
                   
                   
                 E159A 
                 T160A 
                 N161H 
                   
                   
                 S174A 
               
               
                 11 
                   
                   
                   
                   
                 E159C 
                 T160S 
                 N161W 
               
               
                 12 
                   
                   
                   
                   
                 E159C 
                   
                 N161W 
                   
                 M172R 
               
               
                 13 
                   
                   
                   
                   
                 E159R 
                   
                 N161Y 
                   
                   
                   
                   
                 N197K 
                 I199M 
               
               
                 14 
                   
                   
                   
                   
                 E159C 
                 T160S 
                 N161W 
               
               
                 15 
                   
                   
                   
                   
                 E159C 
                   
                 N161W 
               
               
                 16 
                   
                   
                   
                   
                 E159C 
                 T160S 
                 N161W 
               
               
                 17 
                   
                   
                   
                   
                 E159S 
                   
                 N161W 
                   
                   
                   
                   
                 N197K 
                 I199M 
               
               
                 18 
               
               
                 19 
               
               
                 20 
                   
                 N138D 
               
               
                 21 
               
               
                 22 
               
               
                 23 
                 S137R 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 K225H 
                 C231Y 
                   
                   
                   
                   
                   
                   
                   
                 Y357C 
                   
                   
               
               
                 K225H 
                   
                   
                   
                   
                   
                   
                   
                   
                 Y357H 
               
               
                 K225H 
                   
                   
                   
                   
                   
                   
                   
                   
                 Y357C 
               
               
                 K225H 
                   
                   
                   
                   
                   
                   
                   
                   
                 Y357C 
               
               
                 K225H 
               
               
                   
                   
                 I232T 
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
                 I232T 
                   
                   
                   
                   
                 Q354R 
                 Y355H 
                   
                   
                 L404* 
               
               
                   
                   
                 I232T 
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
                 I232T 
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
                 I232T 
                   
                   
                 D312N 
                   
                 Q354R 
                 Y355H 
                   
                 A389T 
               
               
                 K225L 
                 C231R 
                   
                   
                   
                   
                   
                   
                   
                 Y357S 
               
               
                 K225L 
                 C231R 
                   
                   
                   
                   
                   
                   
                   
                 Y357C 
               
               
                 K225H 
                   
                   
                   
                 K311E 
               
               
                 K225L 
                 C231R 
                   
                   
                   
                   
                   
                   
                   
                 Y357C 
               
               
                 K225L 
                 C231R 
                   
                   
                   
                   
                   
                   
                   
                 Y357C 
               
               
                 K225L 
                 C231R 
               
               
                 K225H 
               
               
                 K225L 
                   
                 I232T 
               
               
                 K225L 
                   
                 I232T 
                 A266T 
               
               
                 K225L 
                   
                 I232T 
                   
                   
                   
                 K329N 
               
               
                 K225L 
                   
                 I232T 
               
               
                 K225L 
                   
                 I232T 
               
               
                 K225L 
                   
                 I232T 
               
               
                   
               
               
                 Unique amino acid sequences of BoNT E PACE 3 variants are provided in SEQ ID NOs: 50-68. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 11 
               
               
                   
               
               
                 BoNT E PACE 4 Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
               
                 1 
                   
                   
                 K22R 
                   
                 E28K 
               
               
                 2 
               
               
                 3 
               
               
                 4 
                   
                   
                 K22R 
                   
                 E28K 
               
               
                 5 
                   
                   
                 K22R 
                   
                 E28K 
               
               
                 6 
                   
                   
                 K22R 
                   
                 E28K 
               
               
                 7 
               
               
                 8 
               
               
                 9 
                   
                   
                   
                   
                 E28K 
               
               
                 10 
                   
                   
                   
                   
                 E28K 
                   
                   
                   
                 D128G 
               
               
                 11 
                   
                 I21M 
                   
                   
                 E28K 
               
               
                 12 
                   
                 I21M 
                   
                   
                 E28K 
               
               
                 13 
                   
                   
                   
                   
                 E28K 
               
               
                 14 
                   
                   
                   
                 Q27H 
                   
                 S99A 
                 G101S 
               
               
                 15 
                   
                   
                   
                   
                 E28K 
                   
                   
                 T119K 
               
               
                 16 
               
               
                 17 
               
               
                 18 
               
               
                 19 
               
               
                 20 
               
               
                 21 
               
               
                 22 
               
               
                 23 
               
               
                 24 
                   
                   
                   
                   
                 E28K 
               
               
                 25 
                   
                   
                   
                   
                 E28K 
               
               
                 26 
                   
                   
                   
                   
                 E28K 
               
               
                 27 
                   
                   
                   
                   
                 E28K 
               
               
                 28 
                   
                   
                   
                   
                 E28K 
               
               
                 29 
                   
                   
                   
                   
                 E28K 
               
               
                 30 
                 S6G 
                   
                   
                   
                 E28K 
               
               
                 31 
                   
                   
                   
                   
                 E28K 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
               
                 1 
                 E154G 
                   
                   
                   
                   
                 S163R 
               
               
                 2 
                 E154G 
                   
                 T160A 
               
               
                 3 
                 E154G 
                   
                 T160A 
               
               
                 4 
                 E154G 
                   
                   
                   
                   
                 S163R 
               
               
                 5 
                 E154G 
                   
                   
                   
                   
                 S163R 
               
               
                 6 
                 E154G 
                   
                   
                   
                   
                 S163R 
               
               
                 7 
                 E154G 
                   
                 T160A 
               
               
                 8 
                 E154G 
                   
                   
                   
                 S162A 
                   
                 M172R 
               
               
                 9 
                   
                 E159A 
                 T160A 
                 N161H 
                   
                   
                   
                 S174A 
               
               
                 10 
                   
                 E159A 
                 T160A 
                 N161H 
                   
                   
                   
                 S174A 
               
               
                 11 
                   
                 E159A 
                 T160A 
                 N161H 
                   
                   
                   
                 S174A 
               
               
                 12 
                   
                 E159A 
                 T160A 
                 N161H 
                   
                   
                   
                 S174A 
               
               
                 13 
                   
                 E159A 
                 T160A 
                 N161H 
                   
                 S163R 
                   
                 S174A 
               
               
                 14 
                   
                 E159L 
                   
                 N161Y 
                 S162Q 
               
               
                 15 
                   
                 E159A 
                 T160A 
                 N161H 
                   
                   
                   
                 S174A 
               
               
                 16 
                   
                 E159C 
                   
                 N161W 
               
               
                 17 
                   
                 E159C 
                 T160S 
                 N161W 
               
               
                 18 
                   
                 E159C 
                 T160S 
                 N161W 
               
               
                 19 
                   
                 E159C 
                   
                 N161W 
               
               
                 20 
                   
                 E159C 
                 T160S 
                 N161W 
               
               
                 21 
                   
                 E159C 
                 T160S 
                 N161W 
               
               
                 22 
                   
                 E159C 
                   
                 N161W 
               
               
                 23 
                   
                 E159C 
                 T160S 
                 N161W 
               
               
                 24 
               
               
                 25 
               
               
                 26 
               
               
                 27 
               
               
                 28 
               
               
                 29 
               
               
                 30 
               
               
                 31 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 K225H 
                   
                   
                   
                   
                   
                   
                   
                 I352A 
                   
                   
                 Y357C 
                   
                   
               
               
                   
                   
                 K225H 
                 C231R 
                   
                   
                   
                 I263V 
                   
                   
                   
                   
                   
                 Y357C 
               
               
                   
                   
                 K225H 
                 C231R 
                   
                   
                   
                 I263V 
                   
                   
                   
                   
                   
                 Y357C 
               
               
                   
                   
                 K225H 
                   
                   
                   
                   
                   
                   
                   
                 I352A 
                   
                   
                 Y357C 
               
               
                   
                   
                 K225H 
                   
                   
                   
                   
                   
                   
                   
                 I352A 
                   
                   
                 Y357C 
               
               
                   
                   
                 K225H 
                   
                   
                   
                   
                   
                   
                   
                 I352A 
                   
                   
                 Y357W 
               
               
                   
                   
                 K225H 
                 C231R 
                   
                   
                   
                 I263V 
                   
                   
                   
                   
                   
                 Y357C 
                 I399S 
               
               
                   
                   
                 K225H 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
                   
                   
                 L404* 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
                   
                   
                 L404* 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
                   
                   
                 L404* 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
                   
                   
                 L404* 
               
               
                   
                   
                   
                   
                   
                 K245R 
                   
                   
                 V265G 
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
                   
                   
                 I232T 
                   
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
                   
                   
                 L404* 
               
               
                   
                   
                 K225L 
                 C231R 
                   
                   
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                 I199M 
                   
                 K225L 
                 C231R 
                   
                   
                   
                   
                 V265G 
                   
                   
                 Q354R 
                   
                 Y357C 
               
               
                   
                   
                 K225L 
                 C231R 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 Y357C 
               
               
                   
                 A224S 
                 K225L 
                 C231R 
                   
                 K245R 
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                 I199M 
                   
                 K225L 
                 C231R 
                   
                   
                   
                   
                 V265G 
                 A313S 
                   
                 Q354R 
                   
                 Y357C 
               
               
                   
                   
                 K225L 
                 C231R 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 Y357C 
                   
                 L404* 
               
               
                   
                   
                 K225L 
                 C231R 
                   
                   
                 I247V 
                   
                 V265G 
                   
                 I352V 
                   
                   
                 Y357C 
               
               
                 I199M 
                   
                 K225L 
                 C231R 
                   
                   
                   
                   
                 V265G 
                   
                   
                   
                   
                 Y357C 
               
               
                   
                   
                 K225L 
                   
                 I232T 
                   
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
                 K225L 
                   
                 I232T 
                   
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
                 K225L 
                   
                 I232T 
                   
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
                 K225L 
                   
                 I232T 
                   
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
                 K225L 
                   
                 I232T 
                   
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
                 K225L 
                   
                 I232T 
                   
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
                 K225L 
                   
                 I232T 
                   
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
                 K225L 
                   
                 I232T 
                   
                   
                   
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
               
               
                 Unique amino acid sequences of BoNT E PACE 4 variants are provided in SEQ ID NOs: 69-89. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 12 
               
               
                   
               
               
                 BoNT E PACE 5 Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
               
                 1 
                 Q27H 
                 I35L 
                   
                 S99A 
                 G101S 
                   
                 E159L 
                 N161Y 
                 S162Q 
                 M172R 
               
               
                 2 
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                   
                 E159L 
                 N161Y 
                 S162Q 
                 M172K 
               
               
                 3 
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                   
                 E159L 
                 N161Y 
                 S162Q 
                 M172K 
               
               
                 4 
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 V132G 
                 E159L 
                 N161Y 
                 S162Q 
                 M172K 
               
               
                 5 
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                   
                 E159L 
                 N161Y 
                 S162Q 
                 M172K 
               
               
                 6 
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                   
                 E159L 
                 N161Y 
                 S162Q 
                 M172K 
               
               
                 7 
                 Q27H 
                   
                 D53Y 
                 S99A 
                 G101S 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                   
                 Variant 
               
               
                   
                   
               
               
                   
                 1 
                 I199V 
                 I232T 
                 N243S 
                 T273I 
                   
                 Q354R 
                 Y355H 
                 L404* 
               
               
                   
                 2 
                   
                 I232T 
                   
                   
                 I302M 
                 Q354R 
                 Y355H 
               
               
                   
                 3 
                   
                 I232T 
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 4 
                   
                 I232T 
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 5 
                   
                 I232T 
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 6 
                   
                 I232T 
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                 7 
                   
                 I232T 
                   
                   
                   
                 Q354R 
                 Y355H 
               
               
                   
                   
               
               
                   
                 Unique amino acid sequences of BoNT E PACE 5 variants are provided in SEQ ID NOs: 90-93. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 13 
               
               
                   
               
               
                 BoNT E PACE 6 Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
               
                 1 
                   
                 Q27H 
                 F29L 
                   
                 S99A 
                 G101S 
                 N118D 
                 G127S 
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 2 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 3 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 4 
                 C26Y 
                 Q27H 
                   
                 Y68H 
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 5 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 6 
                   
                 Q27H 
                   
                   
                 S99T 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 7 
                 C26Y 
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 8 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 9 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 10 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 11 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 12 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 13 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 14 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 15 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                 16 
                   
                 Q27H 
                   
                   
                 S99A 
                 G101S 
                 N118D 
                   
                 E159L 
                 N161Y 
                 S162Q 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                 Variant 
               
               
                   
                   
               
               
                   
                 1 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 2 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 3 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 4 
                 S163R 
                 M172K 
                 I232T 
                 N238S 
                   
                 Q354R 
                 Y357H 
               
               
                   
                 5 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
                   
                 I409T 
               
               
                   
                 6 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
                 L404* 
               
               
                   
                 7 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357H 
               
               
                   
                 8 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 9 
                 S163R 
                 M172K 
                 I232T 
                   
                 Q295R 
                 Q354R 
                 Y357P 
               
               
                   
                 10 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357H 
               
               
                   
                 11 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 12 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 13 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 14 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 15 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                 16 
                 S163R 
                 M172K 
                 I232T 
                   
                   
                 Q354R 
                 Y357P 
               
               
                   
                   
               
               
                   
                 Unique amino acid sequences of BoNT E PACE 6 variants are provided in SEQ ID NOs: 94-100. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 14 
               
               
                   
               
               
                 BoNT F PACE 1 (1131) Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
               
                 1 
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                 R240L 
                   
                 F360L 
               
               
                 2 
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                 R240L 
                   
                 F360L 
               
               
                 3 
                   
                   
                   
                   
                 S166Y 
                   
                 N184K 
                   
                 R240L 
                   
                 F360L 
               
               
                 4 
                   
                   
                   
                   
                 S166Y 
                   
                 N184K 
                   
                 R240L 
                   
                 F360L 
               
               
                 5 
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                 R240L 
                   
                 F360L 
               
               
                 6 
                   
                   
                 K146R 
                 N165S 
                 S166Y 
                   
                   
                   
                 R240L 
                   
                 F360L 
               
               
                 7 
                   
                 Y113D 
                 K146R 
                   
                 S166Y 
                   
                   
                   
                 R240L 
                   
                 F360L 
               
               
                 8 
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                 R240L 
                   
                 F360L 
               
               
                 9 
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                 R240L 
                 S350G 
                 F360L 
               
               
                 10 
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                 R240L 
                 S350G 
                 F360L 
               
               
                 11 
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                 R240L 
                 S350G 
                 F360L 
               
               
                 12 
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                 R240L 
                 S350G 
                 F360L 
               
               
                 13 
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                 R240L 
                 S350G 
                 F360L 
               
               
                 14 
                   
                   
                   
                   
                 S166Y 
                   
                 N184S 
                   
                 R240L 
                 S350G 
                 F360L 
               
               
                 15 
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                 R240L 
                 S350G 
                 F360L 
               
               
                 16 
                   
                   
                   
                   
                 S166Y 
                 D175A 
                 N184K 
                   
                 R240L 
                   
                 F360L 
               
               
                 17 
                   
                   
                   
                   
                 S166Y 
                   
                 N184K 
                   
                 R240L 
                   
                 F360L 
               
               
                 18 
                   
                   
                   
                   
                 S166Y 
                   
                 N184K 
                 D185A 
                 R240L 
                   
                 F360L 
               
               
                 19 
                 A82V 
                   
                   
                   
                 S166Y 
                   
                 N184K 
                   
                 R240L 
                   
                 F360L 
               
               
                 20 
                 A82V 
                   
                   
                   
                 S166Y 
                   
                 N184K 
                   
                 R240L 
                   
                 F360L 
               
               
                 21 
                   
                   
                   
                   
                 S166Y 
                   
                 N184K 
                   
                 R240L 
                   
                 F360L 
               
               
                 22 
                   
                   
                   
                   
                 S166Y 
                   
                 N184K 
                   
                 R240L 
                   
                 F360L 
               
               
                 23 
                   
                   
                   
                   
                 S166Y 
                   
                 N184K 
                   
                 R240L 
                   
                 F360L 
               
               
                 24 
                   
                   
                   
                   
                 S166Y 
                   
                 N184K 
                   
                 R240L 
                   
                 F360L 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
               
                 1 
                 Y372H 
                   
                 N396H 
                 P410L 
               
               
                 2 
                 Y372H 
                   
                   
                 P410L 
                 K411N 
               
               
                 3 
                 Y372H 
                   
                   
                 P410L 
               
               
                 4 
                 Y372H 
                   
                   
                 P410L 
               
               
                 5 
                 Y372H 
                   
                   
                 P410L 
               
               
                 6 
                 Y372H 
                   
                 N396H 
                 P410L 
               
               
                 7 
                 Y372H 
                   
                   
                 P410L 
               
               
                 8 
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                   
                   
                   
                 E423K 
               
               
                 9 
                 Y372H 
                 N379D 
                   
                 P410L 
               
               
                 10 
                 Y372H 
                 N379D 
                 N396H 
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 11 
                 Y372H 
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 12 
                 Y372H 
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 13 
                 Y372H 
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 14 
                 Y372H 
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 15 
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 16 
                 Y372H 
                   
                   
                 P410L 
               
               
                 17 
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                   
                   
                   
                 E423K 
               
               
                 18 
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                   
                   
                   
                 E423K 
               
               
                 19 
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                 G420V 
                   
                   
                 E423K 
               
               
                 20 
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                   
                   
                   
                 E423K 
               
               
                 21 
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                   
                   
                   
                 E423K 
               
               
                 22 
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                   
                   
                   
                 E423K 
               
               
                 23 
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                   
                   
                   
                 E423K 
               
               
                 24 
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                   
                   
                   
                 E423K 
               
               
                   
               
               
                 Unique amino acid sequences of BoNT F PACE 1 variants are provided in SEQ ID NOs: 101-116. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 15 
               
               
                   
               
               
                 BoNT F PACE 2 (1207) Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
               
                 1 
                   
                   
                   
                   
                 V155I 
                   
                   
                 S166Y 
                 N184K 
                 R240L 
               
               
                 2 
                   
                   
                 V106A 
                   
                   
                   
                 E164G 
                 S166Y 
                 N184K 
                 R240L 
               
               
                 3 
                 R41H 
                 D60E 
                   
                   
                   
                   
                   
                 S166Y 
                 N184K 
                 R240L 
               
               
                 4 
                   
                   
                   
                   
                   
                   
                   
                 S166Y 
                 N184K 
                 R240L 
               
               
                 5 
                   
                   
                   
                   
                 V155I 
                   
                   
                 S166Y 
                 N184K 
                 R240L 
               
               
                 6 
                   
                   
                 V106A 
                   
                   
                   
                 E164G 
                 S166Y 
                 N184K 
                 R240L 
               
               
                 7 
                   
                   
                   
                   
                 V155I 
                   
                   
                 S166Y 
                 N184K 
                 R240L 
               
               
                 8 
                   
                   
                   
                 Y113H 
                   
                 D161N 
                 E164G 
                 S166Y 
                 N184K 
                 R240L 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                   
                 Variant 
               
               
                   
                   
               
               
                   
                 1 
                   
                 F360L 
                 Y372H 
                   
                 N396H 
                 P410L 
                 K411E 
                 E423K 
               
               
                   
                 2 
                   
                 F360L 
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                 E423K 
               
               
                   
                 3 
                 N339T 
                 F360L 
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                 E423K 
               
               
                   
                 4 
                   
                 F360L 
                 Y372H 
                 I385V 
                 N396H 
                 P410L 
                   
                 E423K 
               
               
                   
                 5 
                   
                 F360L 
                 Y372H 
                   
                 N396H 
                 P410L 
                 K411E 
                 E423K 
               
               
                   
                 6 
                   
                 F360L 
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                 E423K 
               
               
                   
                 7 
                   
                 F360L 
                 Y372H 
                   
                 N396H 
                 P410L 
                 K411E 
                 E423K 
               
               
                   
                 8 
                   
                 F360L 
                 Y372H 
                   
                 N396H 
                 P410L 
                   
                 E423K 
               
               
                   
                   
               
               
                   
                 Unique amino acid sequences of BoNT F PACE 2 variants are provided in SEQ ID NOs: 117-121. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 16 
               
               
                   
               
               
                 BoNT F PACE 3 (1216) Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 1 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                   
                   
               
               
                 2 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 3 
                   
                   
                   
                   
                   
                   
                   
                   
                 N165S 
                 S166Y 
                   
                   
                 N184K 
               
               
                 4 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 5 
                   
                   
                   
                   
                   
                   
                   
                   
                 N165S 
                 S166Y 
                   
                   
                 N184K 
               
               
                 6 
                   
                   
                   
                   
                   
                   
                   
                   
                 N165S 
                 S166Y 
                   
                   
                 N184K 
               
               
                 7 
                 S30N 
                   
                   
                   
                 E66D 
                   
                   
                 Y113D 
                   
                 S166Y 
                   
                   
                 N184K 
                   
                   
                 T214A 
               
               
                 8 
                   
                   
                   
                   
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
               
               
                 9 
                   
                   
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
               
               
                 10 
                   
                   
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 11 
                   
                   
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 12 
                   
                   
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                 S189I 
               
               
                 13 
                   
                   
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                 S167I 
               
               
                 14 
                   
                   
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                 S167I 
               
               
                 15 
                   
                   
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                 S167I 
               
               
                 16 
                   
                   
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 17 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 18 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                 D185A 
               
               
                 19 
                   
                   
                   
                   
                   
                 A82V 
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 20 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 21 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 22 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 23 
                   
                   
                 R41H 
                 D60E 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 24 
                   
                 I39V 
                   
                   
                   
                 A82V 
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                 S350G 
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                 S350G 
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                 S350G 
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                 S350G 
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                 S350G 
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                 F341L 
                   
                 S350G 
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                 S350G 
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                 S350G 
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                 R240L 
                 Y294H 
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                 Y316N 
                   
                 K342R 
                   
                 F360L 
                 Y372H 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425T 
                 V426* 
               
               
                   
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                   
               
               
                   
                 Unique amino acid sequences of BoNT F PACE 3 variants are provided in SEQ ID NOs: 122-137. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 17 
               
               
                   
               
               
                 BoNT F PACE 4 (1279) Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 1 
                   
                   
                   
                   
                 N165S 
                 S166Y 
                   
                   
                 N184K 
                   
                   
                 R240L 
               
               
                 2 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                   
                 R240L 
               
               
                 3 
                   
                   
                   
                 D161N 
                 N165T 
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                 R240L 
               
               
                 4 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                   
                 R240L 
               
               
                 5 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                   
                 R240L 
               
               
                 6 
                   
                   
                   
                   
                 N165S 
                 S166Y 
                   
                   
                 N184K 
                   
                   
                 R240L 
               
               
                 7 
                   
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                 R240L 
               
               
                 8 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                   
                 R240L 
               
               
                 9 
                   
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                 R240L 
               
               
                 10 
                   
                   
                   
                   
                   
                 S166Y 
                 S167I 
                 K172R 
                 N184K 
                   
                   
                 R240L 
               
               
                 11 
                   
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                 R240L 
               
               
                 12 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                   
                 R240L 
               
               
                 13 
                   
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 N184K 
                 P197S 
                   
                 R240L 
               
               
                 14 
                 T79S 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                 R240L 
               
               
                 15 
                   
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                 R240L 
               
               
                 16 
                   
                   
                   
                 D161N 
                   
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                 R240L 
               
               
                 17 
                   
                 V106A 
                   
                 D161N 
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
                 R240L 
               
               
                 18 
                   
                 V106A 
                 S136I 
                 D161N 
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
                 R240L 
               
               
                 19 
                   
                 V106A 
                   
                 D161N 
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
                 R240L 
               
               
                 20 
                   
                 V106A 
                   
                 D161N 
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
                 R240L 
               
               
                 21 
                   
                 V106A 
                   
                 D161N 
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
                 R240L 
               
               
                 22 
                   
                 V106A 
                   
                 D161N 
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
                 R240L 
               
               
                 23 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                   
                 R240L 
               
               
                 24 
                   
                 V106A 
                   
                 D161N 
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
                 R240L 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 T335S 
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                 I370V 
                 Y372H 
                 L375R 
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                 T335S 
                   
                 N358T 
                 F360L 
                   
                 Y372H 
                 L375R 
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                 T335S 
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                 A292S 
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                   
                 D382Y 
                 N396H 
                 P410L 
                 E423K 
               
               
                 K283E 
                   
                   
                   
                 N339T 
                   
                   
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                 A291V 
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                 K347N 
                   
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                 R303H 
                   
                   
                   
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
                 P410L 
                 E423K 
               
               
                   
               
               
                 Unique amino acid sequences of BoNT F PACE 4 variants are provided in SEQ ID NOs: 138-154. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 18 
               
               
                   
               
               
                 BoNT F PACE 5 (1285) Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 1 
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
                   
                 R240L 
                   
                   
               
               
                 2 
                   
                 E121D 
                 D161N 
                 N165T 
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                   
                 R240L 
               
               
                 3 
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
                   
                 R240L 
               
               
                 4 
                   
                 E121D 
                 D161N 
                 N165T 
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                   
                 R240L 
               
               
                 5 
                   
                 E121D 
                 D161N 
                 N165T 
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                   
                 R240L 
               
               
                 6 
                   
                   
                 D161N 
                 N165S 
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                   
                 R240L 
               
               
                 7 
                   
                   
                 D161N 
                 N165T 
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                   
                 R240L 
               
               
                 8 
                   
                   
                 D161N 
                 N165T 
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                   
                 R240L 
               
               
                 9 
                   
                   
                 D161N 
                 N165T 
                 S166Y 
                 S167I 
                 K172R 
                 N184K 
                   
                   
                   
                 R240L 
               
               
                 10 
                   
                 E121K 
                 D161N 
                 N165T 
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                   
                 R240L 
               
               
                 11 
                   
                 E121K 
                 D161N 
                 N165T 
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                   
                 R240L 
               
               
                 12 
                   
                 E121K 
                 D161N 
                 N165T 
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                   
                 R240L 
                   
                 D274E 
               
               
                 13 
                   
                 E121K 
                 D161N 
                 N165T 
                 S166Y 
                 S167I 
                   
                 N184K 
                   
                   
                   
                 R240L 
               
               
                 14 
                   
                 E121K 
                 D161N 
                 N165T 
                 S166Y 
                 S167I 
                   
                 N184K 
                 V193M 
                   
                   
                 R240L 
                 Y253S 
               
               
                 15 
                 V106A 
                   
                 D161N 
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
                   
                 R240L 
               
               
                 16 
                 V106A 
                   
                 D161N 
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
                   
                 R240L 
               
               
                 17 
                 V106A 
                   
                 D161N 
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
                   
                 R240L 
               
               
                 18 
                 V106A 
                   
                 D161N 
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
                 Y210C 
                 R240L 
               
               
                 19 
                 V106A 
                   
                 D161N 
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
                   
                 R240L 
               
               
                 20 
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                   
                   
                 R240L 
               
               
                 21 
                 V106A 
                   
                 D161N 
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
                   
                 R240L 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
                 L375R 
                   
                 F392Y 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
                 L375R 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
                 L375R 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
                 L375R 
                 S389N 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
                 L375R 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
                 L375R 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
                 L375R 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
                 L375R 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
                 L375R 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                   
                   
                 T335S 
                   
                 F360L 
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                   
                   
                   
                 K347N 
                 F360L 
                 Y372H 
               
               
                 T279C 
                   
                   
                   
                 F360L 
                 Y372H 
               
               
                   
                 T299M 
                   
                   
                 F360L 
                 Y372H 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
                   
                   
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
                   
               
               
                   
                 Unique amino acid sequences of BoNT F PACE 5 variants are provided in SEQ ID NOs: 155-170. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 19 
               
               
                   
               
               
                 BoNT F PACE 6 (160508) Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 1 
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                 T214S 
                   
                   
                   
               
               
                 2 
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                 T214S 
               
               
                 3 
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                   
                   
                 Y237S 
               
               
                 4 
                   
                   
                   
                   
                   
                   
                 S166Y 
               
               
                 5 
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
               
               
                 6 
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                 T214S 
               
               
                 7 
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 Y201D 
                 N211D 
               
               
                 8 
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                   
                   
                   
                   
                 T214S 
               
               
                 9 
                   
                   
                   
                   
                   
                   
                 S166Y 
                 S176N 
               
               
                 10 
                 R41H 
                   
                   
                   
                   
                   
                 S166Y 
               
               
                 11 
                 R41H 
                   
                   
                   
                   
                   
                 S166Y 
               
               
                 12 
                   
                   
                   
                   
                   
                   
                 S166Y 
               
               
                 13 
                   
                 D55N 
                 K96N 
                   
                   
                 Y113S 
                 S166Y 
               
               
                 14 
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                 N184H 
               
               
                 15 
                   
                   
                   
                   
                   
                   
                 S166Y 
               
               
                 16 
                   
                   
                   
                   
                   
                   
                 S166Y 
               
               
                 17 
                 R41H 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                   
                 R240L 
               
               
                 18 
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                   
                 R240L 
               
               
                 19 
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                   
                   
                   
                   
                   
                 R240L 
               
               
                 20 
                 R41H 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                   
                 R240L 
                   
                 E259G 
               
               
                 21 
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                   
                 R240L 
                   
                 E259G 
               
               
                 22 
                 R41H 
                   
                   
                   
                 Q109H 
                   
                 S166Y 
                   
                   
                   
                   
                   
                 R240L 
               
               
                 23 
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                   
                 R240L 
               
               
                 24 
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                   
                 R240L 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 T299M 
                   
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 A330E 
               
               
                   
                   
                   
                   
                   
                 N305T 
               
               
                   
                   
                   
                   
                   
                   
                 N329D 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 E338K 
               
               
                 F267L 
               
               
                   
                 I286N 
               
               
                 F267L 
                   
                   
                 R300H 
                 R303H 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                   
                   
                 S415P 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                 S415P 
               
               
                   
                   
                   
                   
                   
                 N379D 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 I416V 
               
               
                   
                   
                   
                   
                   
                   
                   
                 S415P 
               
               
                   
                   
                   
                 Y372H 
                   
                 N379D 
               
               
                   
                   
                   
                   
                   
                 N379D 
               
               
                   
                   
                   
                   
                   
                 N379D 
               
               
                   
                   
                   
                   
                   
                 N379D 
               
               
                   
                   
                   
                   
                   
                 N379D 
               
               
                   
                   
                   
                   
                   
                   
                 D414G 
               
               
                   
                   
                   
                   
                   
                 N379D 
               
               
                   
                   
                   
                   
                 F374L 
                 N379D 
               
               
                   
                   
                   
                   
                   
                 N379D 
               
               
                   
                 F360L 
                   
                 Y372H 
                   
                   
                 D414G 
               
               
                   
                   
                   
                 Y372H 
               
               
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                 F360L 
                   
                 Y372H 
                   
                   
                 D414G 
               
               
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                 F360L 
                 V362A 
                 Y372H 
                   
                   
                 D414G 
               
               
                   
                   
                   
                 Y372H 
               
               
                   
                   
                   
                 Y372H 
               
               
                   
                   
               
               
                   
                 Unique amino acid sequences of BoNT F PACE 6 variants are provided in SEQ ID NOs: 171-189. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 20 
               
             
            
               
                   
               
               
                 BoNT F PACE 7 (160520) Variants 
               
            
           
           
               
               
            
               
                 Variant 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 1 
                   
                   
                   
                 S166Y 
                 N184D 
                   
                   
                   
                   
                   
                   
                 F374C 
                   
               
               
                 2 
                 S100I 
                   
                   
                 S166Y 
               
               
                 3 
                   
                   
                 E121D 
                 S166Y 
                   
                   
                 F267L 
                   
                   
                   
                 D355G 
               
               
                 4 
                   
                   
                   
                 S166Y 
               
               
                 5 
                   
                   
                   
                 S166Y 
               
               
                 6 
                   
                   
                   
                 S166Y 
               
               
                 7 
                   
                 A103V 
                   
                 S166Y 
               
               
                 8 
                   
                   
                   
                 S166Y 
               
               
                 9 
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                   
                 A330E 
               
               
                 10 
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                   
                   
                   
                   
                 F428S 
               
               
                 11 
                   
                   
                   
                 S166Y 
               
               
                 12 
                   
                   
                   
                 S166Y 
               
               
                 13 
                   
                   
                   
                 S166Y 
               
               
                 14 
                   
                   
                   
                 S166Y 
                   
                 P221R 
               
               
                 15 
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                 A307S 
               
               
                 16 
                   
                   
                   
                 S166Y 
                   
                   
                   
                 A292S 
               
               
                   
               
               
                 Unique amino acid sequences of BoNT F PACE 7 variants are provided in SEQ ID NOs: 190-198. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 21 
               
             
            
               
                   
               
               
                 BoNT F PACE 8 (160720) Variants 
               
            
           
           
               
               
            
               
                 Variant 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 1 
                   
                   
                   
                 S166Y 
                 N184D 
                   
                   
                   
                   
                   
                   
                 F374C 
                   
               
               
                 2 
                 S100I 
                   
                   
                 S166Y 
               
               
                 3 
                   
                   
                 E121D 
                 S166Y 
                   
                   
                 F267L 
                   
                   
                   
                 D355G 
               
               
                 4 
                   
                   
                   
                 S166Y 
               
               
                 5 
                   
                   
                   
                 S166Y 
               
               
                 6 
                   
                   
                   
                 S166Y 
               
               
                 7 
                   
                 A103V 
                   
                 S166Y 
               
               
                 8 
                   
                   
                   
                 S166Y 
               
               
                 9 
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                   
                 A330E 
               
               
                 10 
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                   
                   
                   
                   
                 F428S 
               
               
                 11 
                   
                   
                   
                 S166Y 
               
               
                 12 
                   
                   
                   
                 S166Y 
               
               
                 13 
                   
                   
                   
                 S166Y 
               
               
                 14 
                   
                   
                   
                 S166Y 
                   
                 P221R 
               
               
                 15 
                   
                   
                   
                 S166Y 
                   
                   
                   
                   
                 A307S 
               
               
                 16 
                   
                   
                   
                 S166Y 
                   
                   
                   
                 A292S 
               
               
                   
               
               
                 Unique amino acid sequences of BoNT F PACE 8 variants are provided in SEQ ID NOs: 199-208. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 22 
               
             
            
               
                   
               
               
                 BoNT F PACE 9 (160829) Variants 
               
            
           
           
               
               
            
               
                 Variant 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 1 
                   
                 S166Y 
                 K283R 
               
               
                 2 
                   
                 S166Y 
               
               
                 3 
                 N101T 
                 S166Y 
               
               
                   
               
               
                 Unique amino acid sequences of BoNT F PACE 9 variants are provided in SEQ ID NOs: 209-211. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 23 
               
               
                   
               
               
                 BoNT F PACE 10 (2012a) Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 1 
                   
                   
                   
                   
                 V106A 
                 Y113S 
                   
                 S166Y 
                   
                   
                 S176G 
                 N184S 
                 E200K 
                   
                   
               
               
                 2 
                   
                 A63T 
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                   
                 N184K 
                 E200G 
               
               
                 3 
                   
                   
                   
                   
                 V106A 
                 Y113D 
                   
                 S166Y 
                   
                   
                   
                   
                 E200K 
               
               
                 4 
                   
                   
                 E66K 
                   
                 V106A 
                 Y113D 
                   
                 S166Y 
                   
                   
                   
                 N184H 
                 E200K 
               
               
                 5 
                   
                   
                   
                   
                 V106A 
                 Y113S 
                   
                 S166Y 
                   
                   
                 S176G 
                   
                 E200K 
               
               
                 6 
                   
                   
                   
                   
                 V106A 
                 Y113S 
                   
                 S166Y 
                   
                   
                   
                 N184H 
                 E200K 
                 G209D 
               
               
                 7 
                 N11S 
                   
                   
                   
                 V106A 
                 Y113D 
                   
                 S166Y 
                   
                   
                   
                   
                 E200K 
                   
                 T214A 
               
               
                 8 
                   
                   
                   
                   
                 V106A 
                 Y113D 
                 H129Y 
                 S166Y 
                   
                   
                   
                   
                 E200K 
               
               
                 9 
                   
                   
                   
                   
                   
                 Y113D 
                   
                 S166Y 
                   
                   
                   
                 N184H 
                 E200K 
               
               
                 10 
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                   
                   
                   
                 N184S 
                 E200K 
               
               
                 11 
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                   
                   
                   
                 N184S 
                 E200K 
               
               
                 12 
                   
                   
                   
                   
                   
                 Y113D 
                   
                 S166Y 
                 S167C 
                   
                   
                 N184H 
                 E200K 
               
               
                 13 
                   
                   
                   
                   
                   
                 Y113D 
                   
                 S166Y 
                   
                   
                   
                 N184H 
                 E200K 
               
               
                 14 
                   
                   
                   
                   
                   
                 Y113D 
                   
                 S166Y 
                   
                   
                   
                 N184H 
                 E200K 
               
               
                 15 
                   
                   
                   
                 N76D 
                 V106A 
                 Y113D 
                   
                 S166Y 
                   
                   
                   
                 N184H 
                 E200K 
               
               
                 16 
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                   
                   
                   
                 N184S 
                 E200K 
               
               
                 17 
                   
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                 N184K 
                 E200K 
               
               
                 18 
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                   
                   
                   
                 N184K 
                 E200K 
               
               
                 19 
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                   
                   
                   
                 N184K 
                 E200K 
               
               
                 20 
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                   
                   
                   
                 N184K 
                 E200K 
               
               
                 21 
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                   
                   
                   
                 N184K 
                 E200K 
               
               
                 22 
                   
                   
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                   
                 N184K 
                 E200K 
               
               
                 23 
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                   
                   
                   
                 N184K 
                 E200K 
               
               
                 24 
                   
                   
                   
                   
                 V106A 
                   
                   
                 S166Y 
                   
                   
                   
                 N184K 
                 E200K 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 R240L 
                   
                   
                 F267L 
                   
                   
                 F360L 
                 Y372H 
                   
                 P410L 
                   
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
                   
                 P410L 
                 G420V 
               
               
                 R240L 
                 T243A 
                   
                   
                   
                   
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                 G420V 
                   
                 V422I 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
                   
                 P410L 
                   
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
                   
                 P410L 
                   
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
                   
                 P410L 
                 G420V 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
                   
                 P410L 
                 G420V 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 R240L 
                   
                   
                   
                   
                 S350G 
                 F360L 
                 Y372H 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 R240L 
                   
                   
                   
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 R240L 
                   
                   
                   
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 R240L 
                   
                   
                   
                 I277L 
                 S350G 
                 F360L 
                 Y372H 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 R240L 
                   
                   
                   
                   
                   
                 F360L 
                 Y372H 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 R240L 
                   
                   
                   
                   
                 S350G 
                 F360L 
                 Y372H 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 R240L 
                   
                   
                   
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                 R240L 
                   
                 Y244C 
                   
                   
                 S350I 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                 R240L 
                   
                 Y244C 
                   
                   
                   
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                 R240L 
                   
                 Y244C 
                   
                   
                   
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                 R240L 
                   
                 Y244C 
                   
                   
                   
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                 R240L 
                   
                 Y244C 
                   
                   
                   
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                 R240L 
                   
                 Y244C 
                   
                   
                   
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                 R240L 
                   
                 Y244C 
                   
                   
                   
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                 R240L 
                   
                 Y244C 
                   
                   
                   
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                   
                   
                 E423K 
               
               
                   
               
               
                 Unique amino acid sequences of BoNT F PACE 10 variants are provided in SEQ ID NOs: 212-228. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 24 
               
               
                   
               
               
                 BoNT F PACE 11 (2020a) Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
               
                 1 
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
               
               
                 2 
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
               
               
                 3 
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
               
               
                 4 
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
               
               
                 5 
                   
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
               
               
                 6 
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
               
               
                 7 
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
               
               
                 8 
                 E66D 
                   
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
               
               
                 9 
                   
                   
                   
                 V106A 
                   
                 S166Y 
                 S167I 
               
               
                 10 
                   
                   
                   
                 V106A 
                   
                 S166Y 
                 S167I 
               
               
                 11 
                   
                   
                   
                 V106A 
                   
                 S166Y 
                 S167I 
               
               
                 12 
                   
                   
                 N76D 
                 V106A 
                   
                 S166Y 
                 S167I 
               
               
                 13 
                   
                   
                   
                 V106A 
                   
                 S166Y 
                 S167I 
               
               
                 14 
                   
                   
                   
                 V106A 
                   
                 S166Y 
                 S167I 
               
               
                 15 
                   
                   
                   
                 V106A 
                   
                 S166Y 
                 S167I 
               
               
                 16 
                   
                   
                   
                 V106A 
                   
                 S166Y 
                 S167I 
               
               
                 17 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 18 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 19 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 20 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 21 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 22 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 23 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
               
               
                 24 
                   
                 S70F 
                   
                   
                 E164K 
                 S166Y 
                   
                   
                 N184K 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                   
                 Variant 
               
               
                   
                   
               
               
                   
                 1 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                   
                 2 
                   
                 E200G 
                 Y210H 
                   
                   
                 S224I 
                 R240F 
               
               
                   
                 3 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                   
                 4 
                   
                 E200G 
                   
                   
                 E215G 
                   
                 R240L 
               
               
                   
                 5 
                   
                 E200G 
                   
                 T214I 
                   
                 S224I 
                 R240F 
               
               
                   
                 6 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                   
                 7 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                   
                 8 
                   
                 E200G 
                   
                   
                 E215G 
                   
                 R240L 
                 Y244C 
               
               
                   
                 9 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
                 10 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
                 11 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
                 12 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
                 13 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
                 14 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
                 15 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
                 16 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
                 17 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                   
                 18 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                   
                 19 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                   
                 20 
                 Y199H 
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                   
                 21 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                   
                 22 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                   
                 23 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                   
                 24 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 R303H 
                 P309T 
                   
                   
                 F360L 
                   
                 Y372H 
                   
               
               
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                   
                   
                 F360L 
                 K371E 
                 Y372H 
               
               
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                   
                   
                 F360L 
                 K371E 
                 Y372H 
               
               
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                 T335S 
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                 N276T 
                   
                   
                 T335S 
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                 T335S 
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                 T335S 
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                 T335S 
                   
                 F360L 
                   
                 Y372H 
               
               
                 A258S 
                 N276S 
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
               
               
                   
                   
                   
                   
                 T335S 
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                   
                 E423K 
               
               
                   
                   
               
               
                   
                 Unique amino acid sequences of BoNT F PACE 11 variants are provided in SEQ ID NOs: 229-241. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 25 
               
               
                   
               
               
                 BoNT F PACE 11 (2020b) Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 1 
                   
                 E66D 
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                 E200G 
                   
                   
                   
                 R240F 
               
               
                 2 
                   
                 E66D 
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                 E200G 
                   
                   
                 S224I 
                 R240F 
               
               
                 3 
                   
                 E66D 
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                 E200G 
                   
                   
                 S224I 
                 R240F 
               
               
                 4 
                   
                 E66D 
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                 E200G 
                   
                   
                 S224I 
                 R240F 
               
               
                 5 
                   
                 E66D 
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                 E200G 
                   
                   
                 S224I 
                 R240F 
               
               
                 6 
                   
                 E66D 
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                 E200G 
                   
                   
                 S224I 
                 R240F 
               
               
                 7 
                   
                 E66D 
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                 E200G 
                   
                 T214G 
                   
                 R240L 
               
               
                 8 
                   
                   
                   
                   
                 V106A 
                 S166Y 
                 S167I 
                   
                   
                 E200G 
                 N211H 
                   
                 S224I 
                 R240L 
               
               
                 9 
                   
                   
                   
                   
                 V106A 
                 S166Y 
                 S167I 
                   
                   
                 E200G 
                   
                   
                 S224I 
                 R240L 
               
               
                 10 
                   
                   
                   
                   
                 V106A 
                 S166Y 
                 S167I 
                   
                   
                 E200G 
                   
                   
                 S224I 
                 R240L 
               
               
                 11 
                   
                   
                   
                 N76D 
                 V106A 
                 S166Y 
                 S167I 
                   
                   
                 E200G 
                   
                   
                 S224I 
                 R240L 
               
               
                 12 
                   
                   
                   
                   
                 V106A 
                 S166Y 
                 S167I 
                   
                   
                 E200G 
                   
                   
                 S224I 
                 R240L 
               
               
                 13 
                   
                   
                   
                   
                 V106A 
                 S166Y 
                 S167I 
                   
                   
                 E200G 
                   
                   
                 S224I 
                 R240L 
               
               
                 14 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                 E200G 
                   
                   
                 S224I 
                 R240F 
               
               
                 15 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                 E200G 
                   
                   
                   
                 R240F 
               
               
                 16 
                   
                   
                 S70F 
                   
                   
                 S166Y 
                   
                   
                 N184K 
                 E200G 
                   
                   
                 S244I 
                 R240F 
               
               
                 17 
                 A63D 
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                 E200G 
                   
                   
                 S244I 
                 R240L 
               
               
                 18 
                   
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                 E200G 
                   
                   
                   
                 R240F 
               
               
                 19 
                   
                   
                 S70F 
                   
                   
                 S166Y 
                   
                   
                 N184K 
                 E200G 
                   
                   
                 S244I 
                 R240L 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                   
                   
                   
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                   
                 N379D 
               
               
                   
                   
                   
                   
                 F360L 
                 K371E 
                 Y372H 
               
               
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
               
               
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
               
               
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
               
               
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
               
               
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
               
               
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
               
               
                   
                   
                 T335S 
                   
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
                   
                 N396H 
               
               
                   
                 G241S 
                   
                   
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                   
                   
                 N396H 
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
                   
                 N396H 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                   
                 P410L 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 P410L 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 P410L 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 P410L 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 P410L 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 P410L 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 P410L 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 P410L 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 P410L 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 P410L 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 P410L 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 P410L 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 P410L 
                   
                 P410L 
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 P410L 
                   
                   
                   
                   
                   
                 E423K 
               
               
                   
                 P410L 
                 D418Y 
                   
                   
                   
                   
                 E423K 
               
               
                   
                 P410L 
                   
                   
                   
                   
                   
                 E423K 
               
               
                   
                 P410L 
                   
                   
                   
                   
                   
                 E423K 
               
               
                   
                 P410L 
                   
                   
                   
                   
                   
                 E423K 
               
               
                   
                 P410L 
                   
                   
                   
                   
                   
                 E423K 
               
               
                   
                   
               
               
                   
                 Unique amino acid sequences of BoNT F PACE 11 variants are provided in SEQ ID NOs: 242-253. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 26 
               
               
                   
               
               
                 BoNT F PACE 11 (2020c) Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 1 
                 E66D 
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
                   
                   
                   
                   
                 R240L 
               
               
                 2 
                 E66D 
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
                   
                   
                   
                   
                 R240F 
               
               
                 3 
                 E66D 
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
                   
                   
                   
                   
                 R240F 
               
               
                 4 
                 E66D 
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                 5 
                 E66D 
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
                   
                   
                   
                   
                 R240F 
               
               
                 6 
                 E66D 
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                 7 
                 E66D 
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                 8 
                 E66D 
                   
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
                   
                   
                   
                   
                 R240L 
               
               
                 9 
                   
                   
                   
                 Y113D 
                 S166Y 
                 S167C 
                   
                 N184H 
                   
                 E200G 
                   
                   
                   
                   
                 R240L 
               
               
                 10 
                   
                   
                 V106A 
                 Y113D 
                 S166Y 
                 S167I 
                   
                 N184H 
                   
                 E200G 
                   
                 Y210H 
                   
                   
                 R240L 
               
               
                 11 
                   
                   
                   
                 Y113D 
                 S166Y 
                 S167C 
                   
                 N184H 
                   
                 E200G 
                   
                   
                   
                   
                 R240L 
               
               
                 12 
                   
                 N101I 
                   
                   
                 S166Y 
                 S167C 
                   
                 N184H 
                   
                 E200G 
                   
                   
                   
                   
                 R240L 
               
               
                 13 
                   
                   
                 V106A 
                 Y113D 
                 S166Y 
                   
                   
                 N184S 
                   
                 E200G 
                   
                   
                   
                   
                 R240L 
               
               
                 14 
                   
                   
                   
                 Y113D 
                 S166Y 
                 S167C 
                   
                 N184H 
                   
                 E200G 
                   
                   
                   
                   
                 R240L 
               
               
                 15 
                   
                   
                 V106A 
                   
                 S166Y 
                 S167I 
                   
                 N184H 
                   
                 E200A 
                   
                   
                   
                   
                 R240L 
               
               
                 16 
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                 17 
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
                 S207N 
                   
                   
                 S224I 
                 R240F 
               
               
                 18 
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                 19 
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240L 
               
               
                 20 
                   
                   
                 V106A 
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
                   
                   
                 E215K 
                 S224I 
                 R240L 
               
               
                 21 
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                 I190V 
                 E200G 
                   
                   
                   
                 S224I 
                 R240L 
               
               
                 22 
                   
                   
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
                   
                   
                   
                 S224I 
                 R240F 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 F360L 
                 K371E 
                 Y372H 
                   
                   
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                 K371E 
                 Y372H 
               
               
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
                   
                 N379D 
               
               
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                 S350G 
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                 T335S 
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                   
                   
                 F360L 
                   
                 Y372H 
                 L375R 
               
               
                   
                   
                 T335S 
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                   
                 T335S 
                   
                 F360L 
                   
                 Y372H 
               
               
                 R244C 
                   
                   
                   
                 F360L 
                 K371E 
                 Y372H 
               
               
                   
                   
                 T335S 
                   
                 F360L 
                   
                 Y372H 
               
               
                   
                 E310G 
                 T335S 
                   
                 F360L 
                   
                 Y372H 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                   
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                   
                 G420A 
                 L421W 
                 V422L 
                 E423R 
                 I425S 
                 V426* 
               
               
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                   
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                   
                   
                 E423K 
               
               
                   
                 N396H 
                 P410L 
                 D418Y 
                   
                   
                   
                 E423K 
               
               
                   
                   
               
               
                   
                 Unique amino acid sequences of BoNT F PACE 11 variants are provided in SEQ ID NOs: 254-268. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 27 
               
               
                   
               
               
                 BoNT F PACE 11 (2020d) Variants 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Variant 
               
               
                   
               
               
                 1 
                   
                   
                   
                 E66D 
               
               
                 2 
                   
                   
                   
                 E66D 
               
               
                 3 
                   
                   
                   
                 E66D 
               
               
                 4 
                   
                   
                 A63T 
                 E66D 
               
               
                 5 
                   
                   
                   
                 E66D 
               
               
                 6 
                   
                   
                   
                 E66D 
               
               
                 7 
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                 8 
                   
                   
                   
                   
                   
                   
                   
                   
                 Y113D 
               
               
                 9 
                   
                   
                   
                   
                   
                 N76D 
                   
                   
                 Y113D 
               
               
                 10 
                   
                   
                   
                   
                   
                   
                   
                   
                 Y113D 
               
               
                 11 
                   
                   
                   
                   
                   
                   
                   
                 V106A 
                 Y113D 
               
               
                 12 
                   
                 D60Y 
                   
                   
                   
                   
                   
                 V106A 
               
               
                 13 
                   
                   
                   
                   
                   
                   
                   
                   
                 Y113D 
               
               
                 14 
               
               
                 15 
                 N40T 
                   
                   
                   
                   
                   
                 E105A 
               
               
                 16 
                   
                   
                   
                   
                   
                   
                 E105A 
               
               
                 17 
               
               
                 18 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 H129Y 
               
               
                 19 
               
               
                 20 
                   
                   
                   
                   
                 S70F 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                   
                 Variant 
               
               
                   
                   
               
               
                   
                 1 
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
               
               
                   
                 2 
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
               
               
                   
                 3 
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
               
               
                   
                 4 
                 I139V 
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
               
               
                   
                 5 
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
               
               
                   
                 6 
                   
                   
                 S166Y 
                   
                 D175G 
                 N184K 
                   
                 E200G 
               
               
                   
                 7 
                   
                   
                 S166Y 
                   
                   
                 N184S 
                   
                 E200K 
               
               
                   
                 8 
                   
                   
                 S166Y 
                 S167C 
                   
                 N184H 
                   
                 E200K 
               
               
                   
                 9 
                   
                   
                 S166Y 
                 S167C 
                   
                 N184H 
                   
                 E200K 
               
               
                   
                 10 
                   
                   
                 S166Y 
                 S167C 
                   
                 N184H 
                   
                 E200K 
               
               
                   
                 11 
                   
                   
                 S166Y 
                 S167C 
                   
                 N184H 
                   
                 E200K 
               
               
                   
                 12 
                   
                   
                 S166Y 
                   
                   
                 N184S 
                   
                 E200K 
               
               
                   
                 13 
                   
                   
                 S166Y 
                   
                   
                 N184H 
                   
                 E200K 
               
               
                   
                 14 
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
               
               
                   
                 15 
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
               
               
                   
                 16 
                   
                   
                 S166Y 
                   
                   
                 N184K 
                 Y199H 
                 E200K 
               
               
                   
                 17 
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
               
               
                   
                 18 
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
               
               
                   
                 19 
                   
                   
                 S166Y 
                   
                   
                 N184K 
                   
                 E200K 
               
               
                   
                 20 
                   
                 E164K 
                 S166Y 
                   
                   
                 N184K 
                   
                 E200G 
               
               
                   
                   
               
               
                   
                 Unique amino acid sequences of BoNT F PACE 11 variants are provided in SEQ ID NOs: 269-285. 
               
            
           
         
       
     
     Example 5 
     Characterization of Evolved BoNT Proteases 
     This example describes expression and isolation of evolved BoNT F proteases. An expression construct comprising a nucleic acid encoding PACE-2020 BoNT F protease variant L2A was produced. The expression construct also included an N-terminal maltose binding protein (MBP) tag and a poly-histidine C-terminal tag. Transformed cells were subjected to cell disruptor lysis, following by primary purification using Ni-NTA and secondary purification by amylose column (which binds to the MBP). 
       FIGS. 38A-38B  show protease expression and isolation of evolved BoNT proteases.  FIG. 38A  shows a Western blot of evolved BoNT F protease m2020-L2A (“m” indicates a maltose-binding protein tag on the N-terminus of the protein).  FIG. 38B  shows a Western blot of Ni-NTA (top) purified BoNT F proteases m2020-L2A and m2020-L3A and subsequent Amylose-purification of BoNT F proteases m2020-L2A and m2020-L3A. 
     In vitro activity of BoNT F variant 2020-L2A was tested using a VAMP7 cleavage assay.  FIG. 39  shows data indicating that BoNT F variant 2020-L2A protease is active in vitro.  FIG. 40  shows data indicating that the cleavage site of BoNT F variant 2020-L2A protease in VAMP7 has shifted relative to the predicted cleavage site, as measured by MS. 
     PACE experiments using high stringency positive selection were performed in order to improve activity of evolved BoNT F proteases.  FIG. 41  shows the selection strategy used during PACE-3401. Table 29 describes mutations present in clones isolated from PACE-3401. 
     
       
         
           
               
               
               
               
               
               
               
               
               
               
               
               
             
               
                 TABLE 29 
               
               
                   
               
             
            
               
                 L1 
                 a 
                   
                   
                   
                   
                   
                 V106A 
                   
                   
                   
                   
               
               
                   
                 b 
                   
                   
                 Y72H 
                   
                   
                 V106A 
                   
                   
                 S141T 
               
               
                   
                 c 
                   
                   
                 Y72H 
                   
                   
                 V106A 
                   
                   
                 S141T 
               
               
                   
                 d 
                   
                   
                 Y72H 
                   
                   
                 V106A 
                   
                   
                 S141T 
               
               
                   
                 e 
                   
                 K31N 
                 Y72H 
                 N99S 
                   
                 V106A 
               
               
                   
                 f 
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 g 
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 h 
                   
                   
                   
                 N99S 
                   
                 V106A 
               
               
                 L2 
                 a 
                   
                   
                 Y72H 
                   
                   
                 V106A 
                 Y113C 
                 V131G 
                 S141T 
               
               
                   
                 b 
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 c 
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 d 
                 K29E 
                   
                   
                   
                   
                 V106A 
                   
                   
                   
                 I150T 
               
               
                   
                 e 
                   
                   
                   
                 N99S 
                 N101D 
                 V106A 
                   
                   
                   
                 I150T 
               
               
                   
                 f 
                   
                   
                 Y72H 
                   
                   
                 V106A 
                   
                 V131G 
                 S141T 
               
               
                   
                 g 
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 h 
                   
                   
                 Y72H 
                   
                   
                 V106A 
                   
                 V131G 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                 L1 
                 a 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                   
                   
                 E200G 
               
               
                   
                   
                 b 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                 N184T 
                   
                 E200G 
               
               
                   
                   
                 c 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                   
                   
                 E200G 
               
               
                   
                   
                 d 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                 N184T 
                   
                 E200G 
               
               
                   
                   
                 e 
                   
                 S166Y 
                 S167I 
                   
                   
                 G178A 
                   
                   
                 E200G 
               
               
                   
                   
                 f 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                   
                 V193M 
                 E200G 
               
               
                   
                   
                 g 
                   
                 S166Y 
                   
                   
                   
                   
                 N184K 
                   
                 E200G 
               
               
                   
                   
                 h 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                   
                   
                 E200G 
               
               
                   
                 L2 
                 a 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                   
                   
                 E200G 
               
               
                   
                   
                 b 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                   
                 V193M 
                 E200G 
               
               
                   
                   
                 c 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                   
                   
                 E200G 
               
               
                   
                   
                 d 
                   
                 S166Y 
                 S167I 
                 M174T 
                   
                   
                   
                   
                 E200G 
               
               
                   
                   
                 e 
                   
                 S166Y 
                 S167I 
                   
                 G177A 
                   
                   
                   
                 E200G 
               
               
                   
                   
                 f 
                   
                 S166Y 
                 S167I 
                 M174T 
                   
                   
                   
                   
                 E200G 
               
               
                   
                   
                 g 
                 V155I 
                 S166Y 
                 S167I 
                 M174T 
                   
                   
                   
                   
                 E200G 
               
               
                   
                   
                 h 
                   
                 S166Y 
                 S167I 
                   
                   
                   
                   
                 V193M 
                 E200G 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                 S224I 
                 R240L 
                   
                   
                   
                   
                   
                   
               
               
                 Y210H 
                   
                   
                 S224I 
                 R240L 
               
               
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
                   
                 E215G 
                 S224I 
                 R240L 
               
               
                   
                 T214I 
                   
                 S224I 
                 R240L 
               
               
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
                   
                   
                 S224V 
                 R240F 
                   
                   
                   
                 I297L 
                   
                 T335S 
               
               
                   
                   
                 E215G 
                 S224I 
                 R240L 
                 F267I 
                 F270V 
               
               
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
                   
                   
                 S224I 
                 R240L 
                   
                   
                   
                   
                 R303C 
               
               
                   
                   
                   
                 S224I 
                 R240L 
                   
                 F270V 
                 N293D 
               
               
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
                   
                   
                 S224I 
                 R240L 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
                   
               
               
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                   
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                 D418Y 
                 F420S 
                 E423K 
               
               
                   
                   
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                 D418Y 
                   
                 E423K 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                 20(AWLRKSRSSNNGDFQHGLAQP* 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
                   
               
               
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                 S350G 
                 F360L 
                 Y372H 
                 N396H 
                 P410L 
                   
                 420(AWLRKS*) 
               
               
                   
                   
               
            
           
         
       
     
     Luciferase assays were performed to investigate the activity of PACE-3041 BoNT F variants. It was observed that PACE-3041 protease variants have improved apparent activity relative to the parental PACE-2020 protease variants from which they were evolved. For example,  FIG. 42  shows luciferase assay data comparing 2020-L2A and 2020-L3A BoNT F protease-containing phage to PACE-3041 clones. 
       FIG. 43  shows luciferase assay data comparing 2020-L2A and 2020-L3A BoNT F protease-containing phage to PACE-3041 clones isolated from 314-092QS-proB lagoons. 
     It was observed that BoNT F PACE-3041 variants were difficult to express and isolated; however, two clones, 3041-L2F and 3041-L2D were successfully isolated and recombinantly expressed. See,  FIG. 44 . In vitro validation of m3041-L2F was subsequently performed.  FIG. 45  shows data indicating that m3041-L2F retained catalytic activity in vitro. 
     Selectivity of BoNT F 2020 protease variants was also tested. See,  FIG. 46 . Data indicated that off-target cleavage was observed for BoNT F 2020-L2A samples. 
     In order to improve selectivity of BoNT F protease variants, a dual-selection approach was used. Briefly, positive selection for VAMP7 cleavage was combined with negative selection for VAMP1 cleavage (referred to as PACE-2300). Genotypes of BoNT F protease variants isolated from PACE-2300 are shown in Table 29. 
     
       
         
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
                 TABLE 30 
               
               
                   
               
             
            
               
                 L1 
                 a 
                 N6S 
                   
                   
                   
                 I52V 
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
                   
               
               
                   
                 b 
                 N6S 
                   
                   
                   
                   
                   
                 D58Y 
                 E60D 
                   
                   
                 S70H 
                   
                   
                 V106A 
               
               
                   
                 c 
                 N6S 
                   
                   
                   
                   
                   
                   
                   
                 A63V 
                   
                   
                 A63V 
                   
                 V106A 
               
               
                   
                 d 
                 N6S 
                   
                   
                   
                 I52V 
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 e 
                 N6S 
                   
                   
                   
                   
                   
                   
                   
                 A63V 
                   
                   
                 A63V 
                   
                 V106A 
               
               
                   
                 f 
                 N6S 
                   
                   
                   
                   
                   
                   
                   
                 A63V 
                   
                   
                 A63V 
                   
                 V106A 
               
               
                   
                 g 
                 N6S 
                   
                   
                   
                   
                   
                   
                   
                 A63V 
                   
                   
                 A63V 
                   
                 V106A 
               
               
                   
                 h 
                 N6S 
                   
                   
                   
                   
                   
                 D58Y 
                 E60D 
                   
                   
                 S70H 
                   
                 T90I 
                 V106A 
               
               
                 L2 
                 a 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 b 
                   
                 Y10C 
                   
                 R49L 
               
               
                   
                 c 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 d 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 e 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
                 T132I 
               
               
                   
                 f 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 g 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
                 T132I 
               
               
                   
                 h 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
                 T132I 
               
               
                 L3 
                 a 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 b 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 c 
                   
                   
                 D16N 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 d 
                   
                   
                   
                   
                   
                 G53S 
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 e 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 f 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 g 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 h 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                 L4 
                 a 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 b 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 c 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 d 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 e 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 E66K 
                   
                   
                   
                 V106A 
               
               
                   
                 f 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 g 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
                 h 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 V106A 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 L1 
                 a 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                 N211S 
                   
                 S224I 
                   
                   
               
               
                   
                 b 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 c 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 d 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                 N211S 
                   
                 S224I 
               
               
                   
                 e 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 f 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 g 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 h 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                 L2 
                 a 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 b 
                   
                   
                   
                   
                 S166Y 
                   
                 N184K 
                 E200G 
                   
                   
                 S224I 
                 A226S 
               
               
                   
                 c 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 d 
                   
                   
                   
                 D161G 
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 e 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 f 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                 F217L 
                 S224I 
               
               
                   
                 g 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
                   
                 A232T 
               
               
                   
                 h 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                 L3 
                 a 
                   
                   
                 G159S 
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 b 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 c 
                 T123M 
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 d 
                   
                 V145I 
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 e 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
                 A226S 
               
               
                   
                 f 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 g 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
                   
                 A232T 
               
               
                   
                 h 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                 Y201H 
                   
                 S224I 
               
               
                 L4 
                 a 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 b 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 c 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 d 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 e 
                 T123S 
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
                 A226S 
               
               
                   
                 f 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
               
               
                   
                 g 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                   
                   
                 S224I 
                   
                 A232T 
               
               
                   
                 h 
                   
                   
                   
                   
                 S166Y 
                 S167I 
                   
                 E200G 
                 Y201H 
                   
                 S224I 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 R240L 
                   
                   
                   
                 N314S 
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                   
                   
                 N339S 
                 S350G 
                 F360I 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                 N314S 
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                   
                 T335I 
                 N339S 
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240F 
                   
                   
                   
                   
                   
                   
                   
                 T335S 
                   
                   
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                 G325S 
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                 I262T 
                   
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                 S333F 
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240F 
                   
                   
                   
                   
                   
                   
                 S333F 
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                 S333F 
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                 S333F 
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                   
                   
                   
                   
                 S333F 
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                   
                 D274M 
                   
                   
                 D331G 
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                 L264M 
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                 L264M 
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                 L264M 
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240F 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                 L264M 
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                 L264M 
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                 R240L 
                   
                 L264M 
                   
                   
                   
                   
                   
                   
                   
                 S350G 
                 F360L 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
                   
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                 T367S 
                   
                 Y372H 
                   
                   
                 N396Y 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                 F369F 
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 D418Y | | 
                 E423K 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(DWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(DWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                 V377I 
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                 V377I 
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                 V377I 
                   
                 N396H 
                   
                 P410L 
                 420(DWLRKS*) 
               
               
                   
                 T367S 
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                 F369F 
                 Y372H 
                   
                   
                 N396H 
                 N409D 
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                 N409D 
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                 N409D 
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                 N409D 
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                 N409D 
                 P410L 
                 420(DWLRKS*) 
               
               
                   
                 T367S 
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                 F369F 
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                 N379H 
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(AWLRKS*) 
               
               
                   
                   
                   
                 Y372H 
                   
                   
                 N396H 
                   
                 P410L 
                 420(DWLRKS*) 
               
               
                   
                   
               
            
           
         
       
     
     A single clone with apparent activity was isolated from PACE-2300, which was cloned and recombinantly expressed. Luciferase assay data indicated that BoNT F 2300-L3B is active in vitro ( FIG. 48 ). Further in vitro characterization experiments were performed.  FIG. 49  shows data relating to the in vitro characterization of m2300-L3B. m2300-L3B protease retains activity on VAMP7. Improvements in in vitro selectivity for VAMP1/7 for m2300-L3B versus m2020-L2A were observed. Substantial improvements in in vitro selectivity for VAMP1/7 for m2300-L3B versus m2020-L3A were observed.