Abstract:
Methods are disclosed that enable the construction of families of sequences comprising sequence repeats connected as direct fusions or with linkers. Families are constructed from a series of interchangeable cassettes and consist of related sequences that can be easily and efficiently polymerized to form multimers and polymers ranging from 1 to N, where N is theoretically any integer greater than one.

Description:
CROSS REFERENCE TO RELATED APPLICATONS  
       [0001]    This application claims priority to U.S. provisional application number US 60/396,466, filed Jul. 16, 2002, naming Stuart Bussell as inventor. 
     
    
     
       SEQUENCE LISTING  
         [0002]    A sequence listing is provided in electronic and printed form and as an appendix to this application.  
         BACKGROUND  
         [0003]    The present invention relates generally to recombinant DNA technology and recombinant protein expression, and more specifically, to constructs comprising repeat DNA sequences and to methods of making constructs comprising repeat DNA sequences, including constructs that encode polymer peptides and proteins, in which monomers are either fused directly or with linkers.  
           [0004]    Recombinant proteins have become an important class of therapeutics and diagnostics since their introduction in the 1980s. The first recombinant protein therapeutics replaced products isolated from either animal or human tissue. For example, recombinant human growth hormone (recombinant human GH or rhGH) replaced material isolated from the pituitaries of human cadavers (Jorgenson,  Endocrine reviews  12:189, 1991). The need arose because of the transmission of a rare fatal disease, called Creutzfeldt-Jakob disease (CJD), that is transmitted from impurities in pituitary derived hGH. The level of control possible with the recombinant version enabled production of drug certifiably free of known communicable agents.  
           [0005]    Another example of an early recombinant protein is recombinant human insulin (rhI) (Chien,  Drug Development and Industrial Pharmacy  22:753, 1996). In this case, the recombinant product replaced, or supplemented, insulin isolated from the pancreases from swine and cattle. The recombinant protein exactly matches the one found naturally in humans, in contrast with the animal versions that differ by one to three amino acids.  
           [0006]    More recombinant protein therapeutics followed including interferons, interleukins, hematopoetic factors, monoclonal antibodies, and others.  
           [0007]    In the diagnostic field, antibodies, both natural and engineered, are used to recognize and signal the presence of clinical markers. An advantage of engineered antibody fragments over full-length antibodies is that they are amenable to production in facile expression systems such as  E. coli  or  P. pastoris  (Pennell et al.,  Res Immunol  149:599, 1998).  
           [0008]    Some of the in vivo characteristics of recombinant drugs are described by their pharmacokinetic parameters. The field of pharmacokinetics concerns itself with the absorption, distribution, metabolism, and excretion (ADME) of compounds delivered in vivo. Basically, pharmacokinetic parameters describe the concentration of a drug distributed throughout the body over time.  
           [0009]    Generally, absorption of protein drugs requires delivery by injection. A body&#39;s natural barriers tend to prevent the absorption of intact proteins if any other routes of delivery are used. The digestion system breaks down proteins administered orally, while the body&#39;s various epidermal surfaces prevent absorption throughout the body.  
           [0010]    Once injected, proteins tend to distribute throughout the circulatory system where they can react (part of metabolism) with other molecules or undergo excretion. Mathematical models, of varying complexity, are available to explain experimental measurements of drug concentrations as a function of time. One of the basic pharmacokinetic parameters is a drugs half-life, t 1/2 , which is characteristic of the drug&#39;s duration in the bloodstream.  
           [0011]    A key determinant to a protein&#39;s half-life in the blood is its size, and this is a result of elimination of proteins from the blood by glomerular filtration in the kidneys (Venkatachalam et al.,  Circulation Research  43:337, 1978). Basically, the filtration allows proteins smaller than 60 kilodaltons (kD), and other similarly sized molecules, to pass out of the blood, resulting in urinary excretion, while retaining larger ones. This has a major impact on the dosing regimen for a given protein. Proteins smaller than 60 kD tend to need daily, or more frequent, injections.  
           [0012]    One strategy to minimize the discomfort and inconvenience of daily injections is to prolong the action of proteins once introduced in vivo. Two basic strategies are used. One involves the formulation of the protein into a slow release formulation (Putney et al.,  Nature Biotechnology  16:153, 1998). An example of this technique involves formulating proteins into a biocompatible polymer, poly lactic co-glycolytic acid (PLGA), that dissolves slowly over time, releasing protein during the dissolution process. Recombinant hGH is one protein successfully formulated this way (Johnson et al.,  Nature Medicine  2:795, 1996). A disadvantage of this technique that complicates its widespread application is the challenge of formulating and manufacturing each protein so that it is stable during processing and use. Furthermore, injections of PLGA formulated proteins can be uncomfortable.  
           [0013]    The other strategy to prolong a protein&#39;s in vivo action involves modifying the protein so that it acts like a larger particle and is excreted more slowly through the kidneys. While prolonging the proteins in vivo residence, the modification must avoid adverse consequences such as immunogenicity, toxicity, unwanted changes to the molecules distribution, and unwanted changes to its activity.  
           [0014]    A common technique in protein modification involves conjugating a native protein to polyethylene glycol (PEG) or another protein (Roberts et al.,  Adv Drug Deliv Rev  54:459, 2002). PEG molecules are manufactured at all ranges of molecular weights. They can be attached to reactive chemical groups compatible with chemical conjugation to proteins, and they are safe in vivo. Pegylated proteins have been approved for human use. Pegylated interferon is an example (Sharieff et al.,  Cleve Clin J Med  69:155, 2002). Pegylation effectively enhances the size of the resulting conjugate while avoiding immunogenicity or activity alterations. However, PEG has its own chemical and physical characteristics, and this can alter a conjugates ADME. For example, PEG alters the distribution of IL2 in such a way as to unacceptably increase its toxicity (Chen et al.,  The Journal of Pharmacology and Experimental Therapeutics  293:248, 2000). Also, the chemical conjugation is difficult to completely control, and any resulting conjugate is likely to be a mix of chemical species.  
           [0015]    Another promising technique involves conjugating or fusing proteins to a carrier protein. There are many examples of chimeric molecules formed either through chemical reaction between the parent proteins or through the fusion of their gene sequences. In the case of fusion proteins, experience shows that the separate polypeptides constituting a fusion protein generally fold into their three dimensional conformation independently. In fact, often a recombinant protein that misfolds during expression in  E. coli  by itself will fold properly when fused to a protein that regularly folds correctly. Examples include fusions to commercially available proteins such as GST and NusA (see for example Novagen, Madison, Wis.).  
           [0016]    One technique to make therapeutic fusion proteins is to fuse native therapeutics to human serum albumin (HSA) (U.S. Pat. No. 5,876,969). HSA is a 66 kD protein that is abundant in the human bloodstream. It is non-immunogenic and readily available. Potential problems include changed distribution of any resulting conjugate and the effect of HSA as it is shuttled into cells that normally do not contain it intracellularly.  
           [0017]    Another technique is to make therapeutic homomultimer fusion proteins. In this case, the coding DNA sequence for a functional protein is connected to copies of itself. A dimer of superoxide dismutase (“SOD”) is disclosed in U.S. Pat. No. 5,084,390, whereby the hinge region of an immunoglobin joins two copies of the SOD monomer. The resulting dimer has an extended in vivo half-life. In another example, a dimer of erythropoietin is disclosed in U.S. Pat. No. 6,242,570.  
           [0018]    Methods to manufacture highly polymerized sequences, for example polymers having greater than two units, have been developed in the field of artificial protein polymers. Lewis et al ( Protein Expression and Purification  7:400, 1996) reveal a method utilizing compatible, but nonregenerable, overhang restriction sites that are engineered to allow the polymerization of a monomeric spider silk repeating sequence in a geometric fashion. In similar manner, Elmorani, et al. ( Biochemical and Biophysical Research Communication  239:240, 1997) use compatible, but nonregenerable, blunt end restriction sites to produce a polymeric form of wheat gliadin.  
           [0019]    The techniques disclosed in both cases are predicated on the presence of a pair of compatible, nonregenerable, restriction sites at the end of the polymerizing protein sequence. This requirement severely limits the number of sequences that are amenable to polymerization. Another disadvantage of currently available methods is that once a final polymeric sequence is generated, the researchers must employ additional steps to engineer it with the appropriate 5′ and 3′ sequences for expression.  
         SUMMARY OF THE INVENTION  
         [0020]    The present invention provides methods to easily and quickly generate multimers, such as dimers and higher order multimers, of DNA sequences and their open reading frame protein translations, resulting in constructs for the expression of proteins of greater molecular weight and valency. Methods are described whereby a sequence is attached to one or more versions of itself, either via a direct fusion or with a linker, where each version shares strong homology and is generally considered the same via its sequence and mode of action. In addition, the multimer is attached to terminal functional elements. The monomer can theoretically have any sequence and can consist of elements from one or more genes or synthetic DNA fragments. Thus, although the polymerization employs homomultimers, the fundamental monomers themselves can be generated from heterogeneous sequences. Furthermore, heteromultimers can be produced from monomers previously manipulated with the methods of this invention if the constitutive monomers have compatible ends.  
           [0021]    In one aspect, the present invention comprises multimer assemblies of cassettes that comprise nucleic acid sequences having restriction sites that can be ligated together to form constructs (multimer cassettes) having multiple copies of a sequence of interest (the monomer sequence), such as a sequence that encodes a peptide or protein. Restriction sites used to ligate cassettes of a multimer assembly together to make a multimer cassette comprise restriction pair members that when ligated together, do not regenerate a restriction site. In one embodiment of the present invention, multimer assemblies are used that comprise 1) at least one amplification cassette comprising at least a monomer sequence and 2) at least one 3′-terminal cassette comprising at least one 3′ specific sequence or at least one 5′-terminal cassette comprising at least one 5′ specific sequence. Preferably, the 5′-terminal and/or 3′-terminal cassettes additionally comprise at least a portion of the monomer sequence.  
           [0022]    In some preferred embodiments of this aspect of the invention, component cassettes (such as amplification cassettes, 5′-terminal and/or 3′-terminal cassettes) of a multimer assembly can comprise one or more flanking restriction sites that can facilitate cloning of multimer cassettes.  
           [0023]    In some preferred embodiments, component cassettes (such as amplification cassettes, 5′-terminal and/or 3′-terminal cassettes) can comprise one or more linker sequences, such as linker sequences that encode amino acids or peptides that can be used to link monomers. Such linker sequence can also comprise restriction sites, such as restriction pair members that can be used in making multimer cassettes.  
           [0024]    In another aspect, the present invention provides methods of making multimer cassettes. Such methods include ligation of 3′ and 5′ restriction pair members of component cassettes. In some preferred embodiments, the synthesis of multimer cassettes can optionally make use of flanking restriction sites that can be provided in the component cassettes. In some preferred embodiments, the synthesis of multimer cassettes can optionally make use of restriction sites that can be provided in linker sequences included in one or more component cassettes.  
           [0025]    The protein polymers encoded by DNA multimers of a multimer cassette can be expressed in any suitable gene/protein expression system. For example, prokaryotic or eukaryotic systems are suitable, as are in vitro translation systems. The multimer assembly system described here facilitates the multimerization process and enables the production of multimers of any size and with a variety of N-terminal, linker, and C-terminal elements from a limited number of starting DNA sequences. For example, a gene can be designed for intracellular expression with an N-terminal methionine and for extracellular expression by including a secretory signal sequence after the N-terminal methionine.  
           [0026]    The invention can be used to produce constructs having multimeric or polymeric sequences of increased size and multiplicity.  
       
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0027]    [0027]FIG. 1 is a diagram showing an example of a multimer assembly and its cassettes for monomers having a terminal restriction pair. (A) shows a 5′-terminal cassette with sequence elements coding for protein N-terminal elements. The crosshatched elements are restriction sites, the rectangular segments are portions of the monomer sequence, the looping arrows indicate continuation as a plasmid, straight arrows indicate linker sequences, and ˜ refers to arbitrary DNA sequences. The circle is a start codon, and the square is a 5′ specific sequence. Restriction site 1 can include the start codon and/or can be a flanking restriction site for cloning flexibility. Restriction site 3 is the 3′ restriction pair member, and 2 and 4 are flanking restriction sites for cloning flexibility. (B) shows an amplification cassette with sequence elements coding for a polymerizing sequence. Restriction site 5 is the 5′ restriction pair member. (C) shows a 3′-terminal cassette with sequence elements coding for C-terminal elements. The pentagon represents 3′ specific sequence and the hexagon a stop codon. The restriction site arrangement is preferred, but not the only arrangement for construction of an insert cassette. (D) shows one example of a Linker sequence. As shown here, it can contain elements 5′ and 3′ of the restriction pair formed by ligating restriction sites 5 and 3 together. The left and right arrows represent linker 5′ and 3′ elements, respectively.  
         [0028]    [0028]FIG. 2 is a diagram showing one example of a multimer assembly and its cassettes for a monomer with an internal restriction pair. The crosshatched elements are restriction sites, the rectangular segments are portions of the monomer sequence, the looping arrows indicate continuation as a plasmid, straight arrows indicate linker sequences, and ˜ refers to arbitrary DNA sequences. The circle is a start codon, and the square is a 5′ specific sequence. The pentagon represents 3′ specific sequence and the hexagon a stop codon. (A) shows a 5′-terminal cassette with sequence elements coding for N-terminal elements. (B) shows an amplification cassette with sequence elements coding for the polymerizing sequence. The double arrow represents a linker (optional). (C) shows a 3′-terminal cassette with sequence elements coding for C-terminal elements. (D) shows an alternative 3′-terminal cassette that requires use of sequential ligation to form a multimer expression cassette.  
         [0029]    [0029]FIG. 3 is a diagram showing two examples of pathways that can be used in the polymerization of amplification cassettes. Both procedures depicted involve two generalized cassettes, one with insert sequence b1 and the other with insert sequence b2. For pathway A, the b2 containing cassette is opened by digesting with enzymes 1 and 5. The b1 insert sequence is isolated after digesting the b1 containing cassette with enzymes 1 and 3. For pathway B, the b1 containing cassette is opened by digesting with enzymes 2 and 3. The b2 insert sequence is isolated after digesting the b2 containing cassette with enzymes 2 and 5. The final ligations to generate multimer assemblies are similar for both cases. The crosshatched elements are restriction sites, the rectangular segments are insert sequences, the looping arrows indicate continuation as a plasmid, and ˜ refers to arbitrary DNA sequences.  
         [0030]    [0030]FIG. 4 is a diagram showing examples of sequential ligation of cassettes to create a functional multimer cassette of a desired size. The schematic is a generalization of the sequential ligation procedure necessary for use with a 3′-terminal cassette given in FIG. 2D. Pathway A depicts the insertion of an ‘S’ plasmid fragment into a ‘T’ containing plasmid, while Pathway B depicts the insertion of a ‘T’ plasmid fragment into a ‘S’ containing plasmid. In the figure, S+T=5I+AI, AI+3I, 5IAI+3I, or 5I+AI3I, where 5I≡the insert from a 5′-terminal cassette, AI≡the insert from an amplification cassette, 3I≡the insert from a 3′-terminal cassette, 5IAI≡the insert resulting from the ligation of 5I and AI, AI3I≡the insert resulting from the ligation of AI with 3I, and 5IAI3I=≡the insert resulting from the ligation of 5I with AI3I or 5IAI with 3I. Formation of 5IAI3I requires two sequential ligations and generation of intermediate 5IAI or AI3I cassettes for each polymer size made. The crosshatched elements are restriction sites, the rectangular segments are insert sequences, the looping arrows indicate continuation as a plasmid, and ˜ refers to arbitrary DNA sequences.  
         [0031]    [0031]FIG. 5 is a diagram showing possible methods for generation of an insertion cassette. Pathways A and B are alternative pathways for insertion cassette generation based on different arrangements of flanking restriction sites. Pathway A involves opening the 5′-terminal cassette and inserting a fragment from the 3′-terminal cassette, while Pathway B involves opening the 3′-terminal cassette and inserting a fragment from the 5′-terminal cassette. The crosshatched elements are restriction sites, the rectangular segments are portions of the monomer sequence, the looping arrows indicate continuation as a plasmid, straight arrows indicate linker sequences, and ˜ refers to arbitrary DNA sequences. The circle is a start codon, and the square is a 5′ specific sequence. The pentagon represents 3′ specific sequence and the hexagon a stop codon.  
         [0032]    [0032]FIG. 6 is a diagram showing one possible method of generating a functional multimer cassette of a desired size from an insertion cassette and an amplification cassette. The insertion cassette is opened at both sites of the restriction pair with subsequent ligation of the insert from an amplification cassette, but the insert can ligate in the wrong orientation. Correct inserts must be identified by subsequent analysis. The crosshatched elements are restriction sites, the rectangular segments are portions of the monomer sequence, the looping arrows indicate continuation as a plasmid, straight arrows indicate linker sequences, and ˜ refers to arbitrary DNA sequences. The circle is a start codon, and the square is a 5′ specific sequence. The pentagon represents 3′ specific sequence and the hexagon a stop codon.  
         [0033]    [0033]FIG. 7 is a diagram showing another possible method of generating a functional multimer cassette of a desired size from an insertion cassette and an amplification cassette. The insertion cassette is opened with enzymes 3 and 2 to create an oriented ligation, but an additional step is required. In this case, the amplification cassette has flanking restriction site 2 on the 3′ side of restriction site 3. The crosshatched elements are restriction sites, the rectangular segments are portions of the monomer sequence, the looping arrows indicate continuation as a plasmid, straight arrows indicate linker sequences, and ˜ refers to arbitrary DNA sequences. The circle is a start codon, and the square is a 5′ specific sequence. The pentagon represents 3′ specific sequence and the hexagon a stop codon.  
         [0034]    [0034]FIG. 8 is a diagram showing another possible scheme for generating a functional multimer cassette of a desired size from an insertion cassette and an amplification cassette in similar fashion to FIG. 7, but the amplification cassette has flanking restriction site 2 on the 5′ side of restriction site 5.  
         [0035]    [0035]FIG. 9 is a diagram showing the PCR amplification of the hGH gene, its subsequent ligation to generate p0A0, and the ligation of the OmpA leader sequence to generate p0C0A2.  
         [0036]    [0036]FIG. 10 is a diagram showing the PCR mutagenesis of the hGH gene to generate p0A01. The diagram also shows the ligation of the OmpA sequence into p0A01 to generate p0A11A2 and the ligation of the PstI/BamHI fragment from p0A01 into P0A03 to generate p0A11A1.  
         [0037]    [0037]FIG. 11 is a diagram showing the PCR mutagenesis of the hGH gene to generate p0A11B.  
         [0038]    [0038]FIG. 12 is a diagram showing the ligation of synthetic sequences to generate p0A11C1 and p0A11C2.  
         [0039]    [0039]FIG. 13 is diagram showing the polymerization of a GH direct fusion amplification cassette.  
         [0040]    [0040]FIG. 14 is diagram showing the generation of the GH direct fusion insertion cassette, p0A11D, and subsequent ligation of an amplification cassette to generate a multimer expression cassette.  
         [0041]    [0041]FIG. 15 is a diagram showing the PCR mutagenesis of the hGH gene to generate p0A21B, the base amplification cassette for the GH glycine linker assembly.  
         [0042]    [0042]FIG. 16 is a diagram showing the PCR mutagenesis of the hGH gene to generate the base cassettes, p0A31A, p0A31B, and p0A31C, for the GH SWG 4 S assembly.  
         [0043]    [0043]FIG. 17 is a diagram showing the sequential ligation of the GH SWG 4 S assembly cassettes to generate the multimer expression cassette, p0A31E3.  
         [0044]    [0044]FIG. 18 is a picture of an SDS-PAGE gel showing the separation of proteins by molecular weight from separate lysates from cells expressing different polymers of rhGH. Lane 1 contains molecular weight standards, lane 2 the rhGH monomer, lane 3 the rhGH dimer, lane 4 the rhGH trimer, lane 5 the rhGH pentamer, and lane 6 the rhGH nanamer.  
         [0045]    [0045]FIG. 19 is a diagram showing insertion of synthetic sequences to generate the G 4 S assembly 5′-terminal and amplification cassettes.  
         [0046]    [0046]FIG. 20 is a diagram showing PCR mutagenesis of the hGH gene to generate p0A04 and p0A41C.  
         [0047]    [0047]FIG. 21 is a diagram showing ligation of the insert from p0D13A with p0A04 to generate p0A43B and ligation of the PstI/EcoRI fragment from p0A11A1 to generate p0A43A.  
         [0048]    [0048]FIG. 22 is a diagram showing ligations to generate the base cassettes; p0A51A, p0A51B, and p0A51C, for the GH direct fusion assembly utilizing blunt ended HindIII and NcoI sites for the restriction pair.  
         [0049]    [0049]FIG. 23 is a diagram showing the polymerization of the p0A51B insert to generate p0A51B2.  
     
    
     DETAILED DESCRIPTION OF THE INVENTION  
     Introduction  
       [0050]    The current invention discloses methods that extend the polymerization techniques in three important ways. First, it introduces new methods to generate highly polymerized sequences from monomers that are incompatible with previous protein polymerization techniques. Second, it introduces additional linker sequences that, when paired with the monomer sequences, facilitate their use. Third, it introduces methods that facilitate the construction and expression of functional multimers and polymers. Taken together, the new methods enable the generation of large numbers of polymer variants that can differ in sequence and degree of polymerization. These variants can then be tested for desirable traits.  
         [0051]    The disclosed techniques are applicable to any polypeptide sequence and can prove useful for proteins for which increased total molecular weight is deemed advantageous. The disclosed techniques are also useful for proteins for which increased valency is deemed advantageous. For example, expression of single chain antibody fragments fused together as larger multimers have the advantage of high valency and a stable linkage. Furthermore, if cassettes for two different sequences share compatible restriction pair members, they can be co-polymerized to produce heteromultimers.  
       Definitions  
       [0052]    Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Conventional methods are used for these procedures, such as those provided in the art and various general references. Where a term is provided in the singular, the inventors also contemplate the plural of that term. The nomenclature used herein and the laboratory procedures described below are those well known and commonly employed in the art. Where there are discrepancies in terms and definitions used in references that are incorporated by reference, the terms used in this invention shall have the definitions given herein. As employed throughout the disclosure, the following terms, unless otherwise indicated, shall be understood to have the following meanings:  
         [0053]    Monomer. A DNA or amino acid sequence whose polymerization is desirable. A monomer can be a portion of a naturally occurring sequence (for example, a binding domain of an antibody). The sequence can be derived from one or more naturally occurring ones, or can be a synthetic sequence, or can be any combination of sequences of synthetic and natural origins. Monomers of the present invention can comprise linkers. As used herein monomer sequence means a nucleic acid sequence.  
         [0054]    Multimer. A nucleic acid sequence encoding two or more monomers.  
         [0055]    Polymer or Multimeric protein. A functional polypeptide that can be synthesized from a multimer assembly of the present invention. A polymer comprises at least two monomers (where each monomer can optionally comprise one or more linkers), can comprise one or more 5′ translated regions (for example, signal peptides, N-terminal regions, “pro” or “pre” protein sequences, tag sequences, etc.), and can comprise one or more 3′-translated regions (for example, C-terminal regions; tag sequences, etc.)  
         [0056]    Linker. A linker is a DNA or amino acid sequence that connects one DNA sequence with another through covalent bonds or an amino acid or peptide that connects one peptide or protein unit with another peptide or protein unit through peptide bonds. An amino acid or peptide linker can be a single amino acid (for example, glycine) or can be more than one amino acid.  
         [0057]    Restriction Pair. Two restriction sites that have different recognition sequences that are ligation compatible, but when ligated together do not regenerate either of the two original restriction sites. A restriction pair can include two restriction sites that have overhangs, such as BglII and BamHI, or can include any two blunt end restriction sites that do not have the same recognition sequence, such as StuI and NaeI. In a broader application, a restriction pair can also include restriction sites that are initially ligation incompatible but are blunt ended to make them ligation compatible. An example includes blunt ending HindIII and NcoI to make them ligation compatible.  
         [0058]    Restriction pair member or restriction member. A restriction site that is part of a restriction pair. The 5′ and 3′ restriction pair members together make up a restriction pair, and each is the other&#39;s partner.  
         [0059]    5′ restriction pair member or 5′ restriction member or 5′ member. A restriction pair member that is located at the 5′ terminus of a DNA sequence, such as a DNA sequence that, at least in part, encodes a monomer whose multimerization is desired or multimer of the present invention, or is located at the 5′ terminus of a DNA sequence of interest whose ligation to a multimer is desired. The term “5′ restriction pair member” or “5′ member” can be used to refer to an unaltered restriction site (for example, a Bam HI site) or to a restriction site that has been altered, such as, for example, a filled-in 5′ restriction pair member (such as blunt ended Bam HI site), or a fused 5′ restriction pair member (for example, a ligated BamHI/BglII site).  
         [0060]    3′ restriction pair member or 3′ restriction member or 3′ member. A restriction pair member that is located at the 3′ terminus of a DNA sequence, such as a DNA sequence that, at least in part, encodes a monomer whose multimerization is desired or multimer of the present invention, or is located at the 3′ terminus of a DNA sequence of interest whose ligation to a multimer is desired. The term “3′ restriction pair member” or “3′ member” can be used to refer to an unaltered restriction site (for example, a BglII site) or to a restriction site that has been altered, such as, for example, a filled-in 3′ restriction pair member (such as blunt ended BglII site), or a fused 3′ restriction pair member (for example, a ligated BamHI/BglII site).  
         [0061]    Flanking restriction site or flanking site. A restriction site that is not a member of a restriction pair used in the constructs and methods of the present invention. Its location outside of insert sequences and restriction pair members used in the cassettes and methods of the present invention can facilitate manipulation of the insert.  
         [0062]    Insertion restriction site. A specific flanking restriction site that is 3′ of the 3′ restriction pair member of the 5′-terminal cassette and 5′ of the 5′ restriction pair member of the 3′-terminal cassette.  
         [0063]    Amplification cassette. A DNA sequence that includes at least one monomer that is flanked by a restriction pair. An amplification cassette has a 5′ restriction pair member at its 5′ terminus and a 3′ restriction pair member at its 3′ terminus. The restriction pair enables the multimerization of the sequence or the ligation of it to other sequences with ligation compatible restriction sites. An amplification cassette can optionally comprise other sequences as well, such as but not limited to sequences that code for amino acid or peptide linkers.  
         [0064]    5′-terminal cassette. A DNA sequence that comprises a 3′ restriction pair member, at least one 5′-specific sequence, where a 5′-specific sequence is a sequence that, when positioned at the 5′ end of a multimer sequence, can facilitate the use of DNA multimers or the expression, purification, or identification of at least one protein polymer of the present invention, and, preferably, at least a portion of a monomer sequence. The 3′ restriction pair member is ligation compatible with the 5′ terminus of at least one amplification cassette. The 5′-terminal cassette is useful for introducing 5′-terminal DNA sequences that contribute to making a sequence functional. Examples of 5′ specific sequences include, but are not limited to, the translation start codon, secretion sequences, tag sequences, linker sequences, or special restriction sites.  
         [0065]    3′-terminal cassette. A DNA sequence that comprises a 5′ restriction pair member, at least one 3′-specific sequence, where a 3′-specific sequence is a sequence that, when positioned at the 3′ end of a multimer sequence, can facilitate the use of DNA multimers or the expression, purification, or identification of at least one protein polymer of the present invention, and, preferably, at least a portion of a monomer sequence. The 5′ restriction pair member is ligation compatible with the 3′ terminus of at least one amplification cassette. The 3′-terminal cassette is useful for introducing 3′-terminal DNA sequences that contribute to making a sequence functional. Examples of 3′ specific sequences include, but are not limited to, tag sequences, C-terminal sequences, polyadenylation sequences, stop codons, linker sequences, and the like.  
         [0066]    Insert sequence. The functional sequence in a cassette. For the amplification cassette, the functional sequence includes both restriction pair members and all sequence in between, including the monomer sequence. For the 5′-terminal cassette, the functional sequence includes the 3′ restriction pair member, all 5′-specific sequences, and its portion of a monomer sequence, if present. For the 3′-terminal cassette, the functional sequence includes the 5′ restriction pair member, all 3′-specific sequences, and its portion of a monomer sequence, if present. For multimer cassettes, the functional sequence includes the functional sequences of the constitutive cassettes.  
         [0067]    Multimer assembly. The collection of all cassettes that, in combination, after ligation, yields functional multimer DNA sequences or polymer protein sequences of a starting monomer. A multimer assembly comprises one or more 5′-terminal cassettes and one or more amplification cassettes; one or more amplification cassettes and one or more 3′-terminal cassettes; or one or more 5′-terminal cassettes, one or more amplification cassettes, and one or more 3′-terminal cassettes that can be fused using 3′ and 5′ restriction pair members.  
         [0068]    Multimer cassette. A cassette resulting from the ligation of two or more cassettes from the same multimer assembly.  
         [0069]    Insertion Cassette. A multimer cassette generated from the ligation of a 5′-terminal and 3′-terminal cassette of a multimer assembly that is ligation compatible with any of said assembly&#39;s amplification cassettes to generate a multimer cassette.  
         [0070]    Multimer expression cassette. A multimer cassette that, when transcribed and translated in a suitable expression system, produces a polymer protein sequence of a starting monomer.  
         [0071]    Segment of a monomer sequence. A segment of a monomer sequence is a portion of monomer sequence, that is, a nucleic acid sequence that encodes a portion of a monomer.  
       I. Methods of Making Multimer Assemblies  
       [0072]    The present invention includes methods of fusing two or more nucleic acid sequences. The nucleic acid sequences can encode for peptide or protein sequences, such that when the nucleic acid sequences are expressed, a polymeric protein is produced. Preferably, in the methods of the present invention, the peptide or protein monomers encoded by the nucleic acid sequences are identical peptide or protein monomers. However, this is not a requirement of the present invention. The nucleic acid sequence, whose polymerization is desired is called a monomer sequence.  
         [0073]    Monomer sequences can encode proteins or peptides whose function is known or unknown. Preferably, however, the identity and function of the peptide or protein encoded by a monomer sequence is known. Of particular interest are peptides and proteins that can have diagnostic or therapeutic value (for example, human growth hormone, hGH), although the invention is not limited to these protein sequences.  
         [0074]    For example, monomer sequences can encode at least a portion of one or more receptors, receptor ligands, enzymes, inhibitors, transcription factors, translation factors, DNA replication factors, activators, chaperonins, or antibodies. Monomer sequences can also encode at least a portion of one or more cytokines, growth factors, or hormones such as, but not limited to, Interferon-alpha, Interferon-beta, Interferon-gamma, Interleukin-1, Interleukin-2, Interleukin-3, Interleukin-4, Interleukin-5, Interleukin-6, Interleukin-7, Interleukin-8, Interleukin-9, Interleukin-10, Interleukin-11, Interleukin-12, Interleukin-13, Interleukin-14, Interleukin-15, Interleukin-16, Erythropoietin, Colony-Stimulating Factor-1, Granulocyte Colony-stimulating Factor, Granulocyte-Macrophage Colony-Stimulating Factor, Leukemia Inhibitory Factor, Tumor Necrosis Factor, Lymphotoxin, Platelet-Derived Growth Factor, Fibroblast Growth Factors, Vascular Endothelial Cell Growth Factor, Epidermal Growth Factor, Transforming Growth Factor-beta, Transforming Growth Factor-alpha, Thrombopoietin, Stem Cell Factor, Oncostatin M, Amphiregulin, Mullerian-Inhibiting Substance, B-Cell Growth Factor, Macrophage Migration Inhibiting Factor, Endostatin, and Angiostatin. Descriptions of these proteins can be found in Human Cytokines: Handbook for Basic and Clinical Research, Aggarwal, B. B. and Gutterman, J. U. Eds., Blackwell Scientific Publications, Boston, Mass., (1992), which is herein incorporated by reference in its entirety.  
         [0075]    The monomer encoding sequences are polymerized together by ligation of compatible, nonregenerable restriction sites, called restriction pair members. Unlike previous methodologies, the present invention employs cassettes with sequences other than those encoding the original monomer itself in the construction process. For example:  
         [0076]    In the methods of the present invention, multimer assemblies are used that comprise at least one amplification cassette and at least one of the following: at least one 3′-terminal cassette or at least one 5′-terminal cassette. An amplification cassette comprises an insert sequence that includes a monomer sequence whose polymerization is desired, a 5′ restriction pair member at its 5′ terminus, and a 3′ restriction pair member at its 3′ terminus. A 3′-terminal cassette comprises an insert sequence that includes at least one 3′ specific sequence and a 5′ restriction pair member site that can be fused to a 3′ restriction pair member site of at least one of the one or more amplification cassettes. A 5′-terminal cassette, comprises an insert sequence that includes at least one 5′ specific sequence and a 3′ restriction pair member site that can be fused to a 5′ restriction pair member site of at least one of the one or more amplification cassettes. Preferably, the 5′-terminal and/or 3′-terminal cassettes additionally comprise at least a portion of the monomer sequence.  
         [0077]    5′ specific sequences can be, but are not limited to, sequences that enhance transcription, translation, secretion, protein folding, protein solubility, or binding of the protein to specific binding members such as antibodies. 3′ specific sequences can be, but are not limited to, stop codons or sequences that enhance RNA stability, protein folding, protein solubility, or binding of the protein to specific binding members such as antibodies.  
         [0078]    In the multimer assemblies of the present invention, 5′ and 3′ restriction pair members are used to fuse amplification cassettes, and preferably, where applicable, 3′-terminal cassettes to amplification cassettes and 5′-terminal cassettes to amplification cassettes. 5′ and 3′ restriction pair members are preferably unique restriction sites that are ligation compatible, and said ligation destroys each member. In the alternative, 5′ and 3′ restriction pair members can be ligation incompatible sites that are made ligation compatible by blunt ending.  
         [0079]    One aspect of the present invention is construction of cassettes comprising one or more flanking restriction sites that aid their use, but this is not a requirement of the present invention. Preferably, 3′-terminal cassettes and 5′-terminal cassettes, if present, comprise 3′ and 5′ flanking restriction sites. Flanking restriction sites can be any restriction site (except restriction pair member sites used in the same construct), and preferably aid the use of cassettes by increasing the facility of making multimer cassettes. For example, the flanking sites facilitate the manipulation of the insert sequences, including their isolation and ligation. For example, some preferred methods employ an insertion restriction site, which is a specific flanking restriction site that is 3′ of the 3′ restriction pair member of the 5′-terminal cassette and 5′ of the 5′ restriction pair member of the 3′-terminal cassette. Flanking restriction sites can also optionally be used to transfer constructs and assemblies to different expression vectors  
         [0080]    In some preferred methods of the invention, sequences encoding linkers are employed. Multimer assembly cassettes can comprise one or more linker sequences. Multimer assembly cassettes can have linker sequences 5′ of one or more insert sequences, 3′ of one or more insert sequences, or both 5′ and 3′ of one or more insert sequences. Linker sequences can be part of amplification cassettes, 5′-terminal cassettes, 3′-terminal cassettes, or any combination thereof. In preferred aspects of the present invention, nucleic acid sequences that encode amino acid or peptide linkers that are used to link monomers can also comprise restriction sites, such as 3′ or 5′ restriction pair member sites that can facilitate construction of multimer assemblies. This provides a convenient means for introducing restriction pair members for efficient polymerization of monomer sequences through amplification cassettes and optionally 5′-terminal cassette or 3′-terminal cassette ligations. Alternatively, or in addition, amino acid or peptide linkers can be used to provide optimal spacing or folding of translated monomers or a polymer.  
         [0081]    Where more than one linker sequence is used in a single multimer assembly cassette, they may or may not occur between each and every monomer sequence. Where more than one linker sequence is used in a single multimer assembly cassette, they can encode the same or different amino acid or peptide linkers.  
         [0082]    Peptide linkers are well known in the art. Preferably linkers are between one and twenty amino acids in length, and more preferably between one and ten amino acids in length, although length is not a limitation in the linkers of the present invention. Preferably linkers comprise amino acid sequences that do not interfere with the conformation and activity of peptides or proteins encoded by monomers of the present invention. Some preferred linkers of the present invention are those that include the amino acid glycine. Examples include those disclosed in Table 1.  
         [0083]    In an expressed protein polymer, such amino acid or peptide sequences join peptide or protein monomer sequences. If a linker is part of the insert sequence of the amplification cassette, it becomes part of the monomer that is to be multimerized. The linker sequence can comprise at least one restriction pair member.  
         [0084]    The present invention also introduces several methods to expand the use of restriction pair member sites. For example:  
         [0085]    In some methods of the present invention, restriction pair members that are used to join monomer sequences are internal to a monomer sequence. In these embodiments, an amplification cassette comprises a 5′ segment of a monomer sequence and a 3′ segment of a monomer sequence that together comprise the sequence of a complete monomer. The 5′ segment is positioned 3′ of the 3′ segment, the 5′terminus of the 3′ segment is a 5′ restriction pair member, and the 3′ terminus of the 5′ segment is a 3′ restriction pair member. In this case, in making a multimer cassette, ligation of the 3′ restriction pair member of the 5′ segment of one amplification cassette with the 5′ restriction pair member of the 3′ segment of another amplification cassette can form a complete monomer sequence. In order to complete the polymer sequences, a multimer assembly preferably comprises a 5′-terminal cassette that comprises the 5′ monomer segment and a 3′-terminal cassette that comprises the 3′ monomer segment. In this way, monomer sequences provided in the amplification cassettes can be provided in non-contiguous segments. In some preferred methods of the present invention, the amplification cassette further comprises a linker that is positioned between the 5′ segment and the 3′ segment of the monomer sequence.  
         [0086]    In some methods of the present invention, restriction pair members can be overhang restriction sites. In some methods of the present invention, restriction pair members can be blunt end restriction sites. In some other methods of the present invention, restriction pair members are incompatible “overhang” restriction sites that are converted to blunt end restriction sites through the use of polymerases or nucleases.  
         [0087]    In some preferred methods of the present invention, restriction pair members are conveniently provided in one or more linker sequences. In these embodiments, linker sequences comprising a restriction pair member can be engineered onto the 3′, 5′, or both ends of an insert sequence.  
         [0088]    In some preferred methods of the present invention, the 3′-restriction pair member codes for a stop codon that is destroyed upon ligation to the 5′-restriction pair member.  
         [0089]    In one aspect of the present invention, the assembly methodology consists of the following four steps:  
         [0090]    1. Generate or Obtain the DNA for the Monomer.  
         [0091]    Techniques familiar to those skilled in the art include, but are not limited to:  
         [0092]    a. Amplification of a sequence from a DNA library, optionally including any additions or mutations to the sequence in PCR primers.  
         [0093]    b. Chemical synthesis of the sequence  
         [0094]    c. Splicing of sequences together from pre-existing DNA  
         [0095]    2. Decide What Linker Sequence, if any, to Use Between Monomers and Construct a Multimer Assembly.  
         [0096]    Options for the linker include none (direct fusion of monomers), a linker encompassing a restriction pair member within its sequence, a linker with restriction pair members at one or more termini, or a linker lacking a restriction pair member. Once a linker is added, it becomes part of the monomer sequence.  
         [0097]    For each option, three basic cassettes can be generated: one or more 5′-terminal cassettes, at least one amplification cassette, and one or more 3′-terminal cassettes. However, in some instances, all three cassettes are not required. A multimer assembly comprises at least one amplification cassette, and one or more 5′-terminal cassettes or one or more 3′-terminal cassettes, or can have at least one amplification cassette, one or more 5′-terminal cassettes, and one or more 3′-terminal cassettes. In some cases, multiple versions of each cassette may be desirable. Furthermore, the amplification cassette can be polymerized to produce new higher order (multimeric) amplification cassettes.  
         [0098]    The ends of the monomers determine the characteristics of the cassettes. The current invention discloses the use of linkers to introduce ends containing a restriction pair as well the construction of 5′-terminal and/or 3′-terminal cassettes to facilitate their use.  
         [0099]    As an alternative to engineering the ends of a monomer with a restriction pair, then the cassettes can be constructed with a restriction pair internal to the monomer sequence. The construction of the cassettes is modified to accommodate the presence of an noncontiguous monomer in each.  
         [0100]    Finally, a method is disclosed in which the constructions for a restriction pair either at the ends or internal to the monomer is extended to use with a pair of incompatible restriction sites. This method is less preferred, as the method requires that blunt ends for ligation are created for each ligation step (by nuclease digestion or polymerase fill-in, or both), decreasing the efficiency of the procedure.  
         [0101]    The following are the general steps for construction of the assemblies for each possible restriction pair case:  
         [0102]    a. Using a monomer sequence with a terminal restriction pair.  
         [0103]    The scheme shown in FIG. 1 is applicable for any monomer sequence that can be engineered with a terminal restriction pair. The steps to engineer the assembly can include the following:  
         [0104]    (1) Engineer 5′-terminal cassettes containing one or more 5′ specific DNA sequences (for example, start codon, secretion sequence, etc.), preferably the monomer sequence, linker sequence, if present, and the 3′ member of the restriction pair.  
         [0105]    (2) Engineer an amplification cassette containing a 5′ restriction member, optionally a first linker sequence, at least one monomer sequence, optionally a second linker sequence, and a 3′ restriction member.  
         [0106]    (3) Engineer 3′-terminal cassettes containing a 5′ restriction member, optionally a linker sequence, preferably the monomer sequence, and one or more 3′-terminal specific DNA sequences (specific recognition sequences, stop codon, etc.).  
         [0107]    An alternative formulation involves 5′-terminal and/or 3′-terminal cassettes that do not include any monomer sequence. The utility of including the monomer sequence in both terminal cassettes lies in utilizing the restriction pair members to join each terminal cassette to an amplification cassette, however, this is not a requirement of the present invention.  
         [0108]    b. Using a monomer sequence with an internal restriction pair.  
         [0109]    The scheme shown in FIG. 2 is applicable for any monomer sequence that can be engineered with an internal restriction pair. The steps to engineer the assembly include the following:  
         [0110]    (1) Engineer 5′-terminal cassettes containing one or more 5′ specific DNA sequences (start codon, secretion sequence, etc.), the portion of a monomer sequence that occurs on the 5′ side of the restriction pair (the 5′ monomer segment), and finally the 3′ restriction pair member.  
         [0111]    (2) Engineer an amplification cassette containing a 5′ restriction pair member, DNA encoding the portion of a monomer sequence that occurs 3′ of the restriction pair (the 3′ monomer segment), optionally a linker sequence, DNA encoding the portion of a monomer that occurs 5′ of the restriction pair (the 5′ monomer segment), and a 3′ restriction pair member.  
         [0112]    (3) Engineer 3′-terminal cassettes containing the 5′ restriction pair member, the portion of a monomer sequence that occurs 3′ of the restriction pair (the 3′ monomer segment), and one or more 3′-terminal specific DNA sequences (specific recognition sequences, stop codon, etc.).  
         [0113]    c. Using a monomer sequence with a pair of incompatible restriction sites made compatible by blunt ending.  
         [0114]    Either scheme shown in FIG. 1 or FIG. 2 are applicable, but in this case the restriction pair consists of restriction sites that are blunt ended to make them compatible.  
         [0115]    Once constructed, the amplification cassette enables generation of a sequence containing any number of monomers fused together.  
         [0116]    3. Polymerize the Amplification Cassette in an Arithmetic, Geometric, or Mixed Progression (see FIG. 3).  
         [0117]    A series of amplification cassettes are generated from the original amplification cassette. The technique involves digesting a first construct comprising an amplification cassette at two 5′ or two 3′ sites of an insert, one of which is a restriction pair member site and the other of which is an external flanking site (external to the restriction pair member site), to open up the construct. This is followed by digesting a second construct comprising an amplification cassette at the same flanking site, but with the opposite restriction pair member, to release the amplification sequence from the plasmid as a fragment. This sequence is then ligated into the opened first plasmid construct from before. Both restriction sites used in the ligation are destroyed, but the resulting cassette has intact flanking restriction sites and an intact restriction pair on the ends that enable further polymerizations.  
         [0118]    Mixing and matching the cassettes used to open a construct that comprises an amplification cassette and to generate an insert from a construct that comprises an amplification cassette enables new cassettes of any size to be made in an arithmetic, geometric, or mixed progression. For example, if the monomer is used to both open the plasmid and create insert, a dimer cassette is made. If the resulting dimer is used for both, then a tetramer is made. If this tetramer is used for both, then an octamer is made, and continuation leads to a binomial geometric progression. On the other hand, if the monomer is always used as the insert and the newest cassette is used to receive the insert, an arithmetic progression of one is produced. For instance, when a dimer construct is opened and a monomer fragment inserted, then a trimer is produced. When a trimer construct is opened and a monomer fragment is inserted, then a tetramer is produced. In general, any new cassette can be mixed with any previously generated cassette to allow rapid generation of a polymer of any desired size. For example, if a polymer of size 20 is desired, the 16 mer is generated geometrically, and ligating the 16 mer to the tetramer generates the 20 mer in a total of only 5 ligations.  
         [0119]    Subsequent ligation to 5′- and 3′-terminal cassettes can enable production of a functional multimer. The multimer&#39;s size, based on actual molecular weight, is approximately a whole number multiple of the original. In addition, the composition of the multimer is almost identical to the monomer, differing only because of any linker sequences or terminal flanking regions that are used.  
         [0120]    It is important to note that the polymerization does not require flanking sites. Without flanking sites, the ligations can occur with the fragments joined in either orientation, and more laborious subsequent analysis is needed to identify the correct constructs. In contrast, use of flanking sites facilitates the process by enabling oriented ligations.  
         [0121]    4. Ligate the Cassettes Together to Give a Full Length, Functional, Multimer.  
         [0122]    The cassettes can be ligated sequentially as shown in FIG. 4, or an insertion cassette can be created from the 5′- and 3′-terminal cassettes as diagramed in FIG. 5 with subsequent insertion of the polymerized amplification cassette as shown in FIGS. 6, 7, and  8 . The use of an insertion cassette expedites the creation of a series of multimers with the same 5′ and 3′ terminal elements. FIG. 6 illustrates a technique for the ligation of the fragment from an amplification cassette into an insertion cassette using only the restriction pair restriction sites. However, the ligation is not oriented, necessitating additional analysis to identify correct constructs. FIGS. 7 and 8 show equivalent oriented ligations that result from different arrangements of flanking sequences.  
         [0123]    [0123]FIG. 4 illustrates a method of making a multimer cassette from two cassettes from a multimer assembly utilizing flanking sites comprising a first cassette comprising either a 5′-restriction pair member or a 3′-restriction pair member and a second cassette comprising both a 5′-restriction pair member and a 3′-restriction pair member and further comprising:  
         [0124]    1) providing the first cassette with a first flanking restriction site at one end, either 5′ or 3′, of its insert sequence;  
         [0125]    2) providing the second cassette with a second flanking restriction site that is, or is made, ligation compatible with the first flanking site and is on the same side, either 5′ or 3′, of its insert sequence as the first flanking restriction site is relative to the first cassette&#39;s insert sequence;  
         [0126]    3) digesting the first cassette at its restriction pair member and the first flanking site and isolating the first fragment containing the insert sequence;  
         [0127]    4) digesting the second cassette at its restriction pair member partner to the first cassette&#39;s restriction pair member and at the second flanking site and isolating the second fragment containing the insert sequence;  
         [0128]    5) ligating the first fragment with the second fragment to generate a multimer cassette.  
         [0129]    The identities of the first and second cassettes can vary. For example, the first cassette can be a 3′-terminal cassette and the second cassette an amplification cassette, the first cassette can be a 5′ terminal cassette and the second cassette an amplification cassette, the first cassette can be a 3′-terminal cassette and the second cassette a multimer 20 cassette constructed from a 5′-terminal cassette and an amplification cassette, or the first cassette can be a 5′-terminal cassette and the second cassette a multimer cassette constructed from a 3′-terminal cassette and an amplification cassette.  
         [0130]    For the case when the first cassette is a 3′-terminal cassette and the second cassette is an amplification cassette, if the amplification cassette is digested at its 3′ restriction pair member and a flanking restriction site on the 5′ side of its 5′ restriction member to generate a ligatable fragment, then the 3′-terminal cassette is digested at its 5′ restriction pair member and a flanking restriction site on the 5′ side of this member to generate a ligatable cassette. Alternatively, if the amplification cassette is digested at its 3′ restriction pair member and a flanking restriction site on the 3′ side of this member to generate a ligatable cassette, then the 3′-terminal cassette is digested at its 5′ restriction pair member and a flanking restriction site on the 3′ side of its complete insert to generate a ligatable fragment.  
         [0131]    It is important to note that the ligation of cassettes together does not require flanking sites. However, flanking sites enable oriented ligations. For example, if flanking sites are absent, a method of making a multimer cassette from two cassettes from a multimer assembly comprising a first cassette comprising either a 5′-restriction pair member or a 3′-restriction pair member and a second cassette comprising both a 5′-restriction pair member and a 3′-restriction pair member comprises:  
         [0132]    1) digesting the first cassette at its restriction pair member and isolating the first fragment containing the insert sequence;  
         [0133]    2) digesting the second cassette at both its restriction pair member sites and isolating the second fragment containing the insert sequence;  
         [0134]    3) ligating the first fragment with the second fragment and screening for correct ligation orientation to generate a multimer cassette.  
         [0135]    Again, the identities of the first and second cassettes can vary. The first cassette can be a 3′-terminal cassette and the second cassette an amplification cassette, the first cassette can be a 5′-terminal cassette and the second cassette an amplification cassette, the first cassette can be a 3′-terminal cassette and the second cassette a multimer cassette constructed from a 5′-terminal cassette and an amplification cassette, or the first cassette can be a 5′-terminal cassette and the second cassette a multimer cassette constructed from a 3′-terminal cassette and an amplification cassette.  
         [0136]    [0136]FIG. 5 illustrates a method of making an insertion cassette from the 5′-terminal cassette and the 3′-terminal cassette when each shares an insertion restriction site. The method comprises:  
         [0137]    1) providing the 5′-terminal cassette with a first flanking restriction site, independent of the insertion restriction site, that is outside of the sequence including the insert sequence and insertion restriction site of the 5′-terminal cassette;  
         [0138]    2) providing the 3′-terminal cassette with a second flanking restriction site, independent of the insertion restriction site, that is outside of the sequence including the insert sequence and insertion restriction site of the 3′-terminal cassette and is, or is made, ligation compatible with the first flanking site and is on the same side, either 5′ or 3′, of its insert sequence as the first flanking restriction site is relative to the 5′-terminal cassette&#39;s insert sequence;  
         [0139]    3) digesting the 5′-terminal cassette at its insertion restriction site and the first flanking site and isolating the first fragment containing the insert sequence;  
         [0140]    4) digesting the 3′-terminal cassette at its insertion restriction site and the second flanking site and isolating the second fragment containing the insert sequence;  
         [0141]    5) ligating the first fragment with the second fragment to generate an insertion cassette.  
         [0142]    [0142]FIG. 6 illustrates a method of making a multimer cassette comprising an insertion cassette and an amplification cassette from a multimer assembly comprising:  
         [0143]    1) digesting the insertion cassette at both its restriction pair member sites and isolating the first fragment containing the insert sequence;  
         [0144]    2) digesting the amplification cassette at both its restriction pair member sites and isolating the second fragment containing the insert sequence;  
         [0145]    3) ligating the first fragment with the second fragment and screening for correct ligation orientation to generate a multimer cassette.  
         [0146]    [0146]FIGS. 7 and 8 illustrate a method of making a multimer cassette comprising an insertion cassette and an amplification cassette comprising:  
         [0147]    1) digesting the amplification cassette at the insertion restriction site and its restriction pair member on the opposite side, either 5′ or 3′, of the insert sequence and isolating the first fragment containing the insert sequence;  
         [0148]    2) digesting the insertion cassette at the insertion restriction site and the restriction pair member partner to the digested amplification cassette&#39;s restriction pair member and isolating the second fragment containing the insert sequence;  
         [0149]    3) ligating the first fragment with the second fragment to generate a multimer cassette precursor;  
         [0150]    4) digesting the multimer cassette precursor at both restriction pair members, isolating the fragment containing the insert sequence, and ligating it with itself to generate a multimer cassette.  
         [0151]    Once constructed, the gene for the multimer can be used as an insert to construct other cassettes or to express it in a suitable transcription and translation system. Once isolated in the correct conformation and with the necessary degree of purity, polymeric polypeptides are available for applications in the fields of medicine, veterinary care, research and development, diagnostics, etc. The present invention comprises proteins made from multimer assemblies of the present invention.  
         [0152]    Each cassette can involve a fusion of any of a number of functional elements. For example, any construction involving a linker is by nature a heteromultimer, because the monomer contains at least two functional elements. A particularly expeditious method to produce these fusions is to treat each functional element as a nested assembly. In other words, each element itself is an assembly that consists of individual cassettes.  
         [0153]    The current methods are easily extended to heteromultimers if two sequences share compatible restriction sites. For instance, two distinct monomer amplification cassettes, A and B, can be ligated together if they share the same restriction pair. Subsequent polymerization of this new “monomer” results in an alternating sequence, ABAB . . . . Any pattern of alternating sequences can theoretically be constructed from any number of initial monomers. For example, the pattern ABBCABBC . . . is just one possibility.  
       II Multimer Assemblies and Multimer Cassettes  
       [0154]    The present invention includes multimer assemblies made using the methods of the present invention and novel cassettes incorporating novel restriction pair members. In some preferred aspects of the present invention, a multimer assembly of the present invention comprises two or more amplification cassettes, in which fused 5′ and 3′ restriction pair member sites join the amplification cassettes. An amplification cassette can comprise any practical number of monomer sequences.  
         [0155]    Multimer assemblies of the present invention comprise component constructs having 5′ restriction pair members, 3′ restriction pair members, or both 5′ restriction pair members and 3′ restriction pair members that can be used to make multimer cassettes, including multimer expression cassettes. Such cassettes are synthesized by joining component cassettes (such as 5′-terminal cassettes, 3′-terminal cassettes, and amplification cassettes) by ligating a 3′ restriction pair member site of one component cassette to a 5′ restriction pair member site of another component cassette.  
         [0156]    One multimer assembly of the present invention comprises one or more amplification cassettes and at least one 3′-terminal cassette. Another multimer assembly of the present invention comprises one or more amplification cassettes and at least one 5′-terminal cassette. Another multimer assembly of the invention comprises one or more amplification cassettes, at least one 3′-terminal cassette, and at least one 5′-terminal cassette.  
         [0157]    Multimer expression cassettes made from multimer assemblies of the present invention include, for example, multimer cassettes in which a 5′-terminal cassette is fused to an amplification cassette comprising a single monomer, multimer cassettes in which a 5′-terminal cassette is fused to a multimer amplification cassette constructed from multiple amplification cassettes, and multimer cassettes in which a 5′-terminal cassette is fused to a multimer cassette comprising one or more amplification cassettes and at least one 3′-terminal cassette. Multimer expression cassettes made from multimer assemblies of the present invention also include, for example, multimer cassettes in which a 3′-terminal cassette is fused to an amplification cassette, multimer cassettes in which a 3′-terminal cassette is fused to a multimer amplification cassette constructed from multiple amplification cassettes, and multimer cassettes in which a 3′-terminal cassette is fused to a multimer cassette comprising one or more amplification cassettes and at least one 5′-terminal cassette.  
         [0158]    The present invention also includes novel amplification cassettes. In one aspect of the present invention, an amplification cassette comprises at least one linker, in which at least one of the one or more linkers comprises at least one restriction pair partner. Amplification cassettes can be fused using restriction pair partners, at least one of which is introduced in the linker, to form a multimer amplification cassette. The method of making the multimer amplification cassette is by joining two or more amplification cassettes by ligating the first restriction pair partner of at least one of the two or more amplification cassettes to the second restriction pair partner of at least one other of the two or more amplification cassettes to generate a multimer cassette. The present invention includes multimer amplification cassettes comprising component amplification cassettes that incorporate linkers, and multimer assemblies and multimer expression cassettes that include such multimer amplification cassettes.  
         [0159]    Also included as amplification cassettes of the present invention are amplification cassettes that comprise monomer sequences in noncontiguous orientation. For example, an amplification cassette can comprise a 5′ segment of a monomer sequence and a 3′ segment of a monomer sequence that together comprise the sequence of a complete monomer, in which the 5′ segment is positioned 3′ of the 3′ monomer segment. In these embodiments, the 5′terminus of the 3′ monomer segment is preferably a 5′ restriction pair member and the 3′ terminus of the 5′ monomer segment is preferably a 3′ restriction pair member. The present invention also includes multimer amplification cassettes comprising two or more amplification cassettes that comprise monomer sequence in noncontiguous orientation. Such multimer cassettes comprising multiple amplification cassettes can be made by ligating a 3′ restriction member of at least one of the two or more amplification cassettes to a 5′ restriction member of at least one other of the two or more amplification cassettes. The present invention also includes multimer assemblies and multimer expression cassettes that include such amplification and multimer amplification cassettes.  
         [0160]    In yet another aspect, the present invention includes amplification cassettes that comprise 3′ and 5′ restriction pair members comprising restriction sites that are initially ligation incompatible but are blunt ended to make them ligation compatible. The present invention also includes multimer amplification cassettes comprising two or more amplification cassettes that comprise noncompatible sites that have been blunt-ended and then ligated to join the two or more amplification cassettes. The present invention also includes multimer assemblies and multimer expression cassettes that include such amplification and multimer amplification cassettes.  
         [0161]    The invention includes multimer assembly cassettes in vectors, including cloning and expression vectors, where expression vectors can be designed for in vitro or in vivo expression. The vectors can be designed for in vivo expression in prokaryotes or eukaryotes, including but not limited to, bacterial cells, fungal cells, algal cells, plant cells, insect cells, avian cells, and mammalian cells. The present invention also encompasses cells that include such vectors and polymeric proteins made using vectors that comprise multimeric expression vectors of the present invention. The present invention also encompasses polymeric proteins expressed from the multimeric assemblies of the present invention.  
         [0162]    The disclosed invention also encompasses the construction of different multimer assemblies involving multimeric hGH, and multimer cassettes made using the methods of the present invention that comprise multimerized hGH sequences or multimerized portions of hGH. Sequences encoding hGH or portions thereof that are part of multimer cassettes and multimer assemblies of the present invention include sequences that encode hGH taking into account the redundancy of the genetic code. Sequences encoding hGH or portions thereof that are part of multimer cassettes and multimer assemblies of the present invention include sequences that encode hGH can also comprise sequence changes with respect to the human GH gene sequence that change the amino acid sequence where such changes do not detrimentally affect the activity of the protein or portion thereof.  
         [0163]    The hGH assemblies can differ in the functional elements included, such as those provided by 3′- or 5′-terminal elements. The ease of producing these assemblies, and the resulting multimers and polymers, demonstrates the utility of the methods disclosed. In the examples below, restriction sites outside, and flanking, the restriction pair sites are engineered in order to facilitate the manipulation of the cassettes.  
         [0164]    Endogenous hGH appears in several forms in vivo as a result of expression from more than one gene, as well as alternative gene splicing. The predominant mature form of hGH is a single polypeptide chain consisting of 191 amino acids. The DNA and protein sequences for this predominant form are given as SEQ ID NO: 1 and SEQ ID NO: 2, respectively.  
         [0165]    In the following paragraphs, the term “engineer” refers to using standard techniques of molecular biology generally known to those skilled in the art. Standard techniques include, but are not restricted to, restriction digestion and ligation, PCR amplification and mutagenesis, DNA synthesis, DNA isolation and purification, etc., as described in Sambrook et al. (2000), which are hereby incorporated by reference. As such, the details are only described if they bear directly on the present invention or deviate from common practice.  
       EXAMPLES  
       [0166]    A drawback to rhGH therapy is the need for once daily injections. Understandably, patient preference is for a minimum of injections. In an attempt to overcome this, rhGH has been formulated with PLGA in microspheres, chemically linked to PEG, and fused to HSA in order to produce longer acting versions. Here we describe the construction of families of multimeric rhGHs, according to the steps below using the general procedures shown in FIGS.  1  to  8 .  
       Example 1  
       [0167]    The first example involves isolation of the GH gene. Steps to isolate the hGH gene are summarized in FIG. 9. hGH is highly expressed in the anterior pituitary gland. As a result, mRNA of hGH is abundantly found in lysates of human pituitary. The gene for hGH is PCR amplified from human pituitary cDNA (Human Pituitary Gland Quick-CloneTM cDNA, BD Biosciences Clontech, Palo Alto, Calif., catalog #7173-1) using SEQ ID NO: 3 as the 5′ primer and SEQ ID NO: 4 as the 3′ primer. The 5′ primer has an NdeI restriction enzyme site coding for an N-terminal methionine, and the 3′ primer has a BamHI restriction enzyme site immediately after the TAG stop codon. The resulting PCR fragment is isolated from the reaction mix using standard techniques, as are all subsequent ones.  
         [0168]    The purified PCR fragment is ligated into parent plasmid pET41a (Novagen, Madison, Wis.) after both insert and plasmid are digested with NdeI and BamHI and purified, again using standard techniques. This plasmid ligation mixture, and all others unless otherwise indicated, is transformed into DH5α cells and plated on LB/antibiotic plates. Single colonies are sub-cultured and plasmid DNA is isolated from each. Restriction enzyme analysis is used to confirm the presence of an insert into the plasmid, and plasmids with insert are sent for DNA sequencing using SEQ ID NO: 5 and SEQ ID NO: 6 (Novagen, Madison, Wis.) as amplification primers for the 5′ and 3′ ends, respectively. Plasmid with correct insert is identified as p0A0, and the DNA coding region and corresponding open reading frame (ORF) translation are listed in SEQ ID NO: 7 and SEQ ID NO: 8, respectively. The convention for the sequences is that the restriction sites are included at the termini of DNA sequences and only translated amino acids that eventually appear in an expressed insert are given. Expression of protein from p0A0 yields a 192 amino acid protein consisting of full length hGH with an additional N-terminal methionine.  
         [0169]    It is convenient to engineer a high copy number plasmid that contains the hGH gene and enables digestion of the hGH gene in its interior so that 5′ or 3′ elements can be swapped in and out. The gene for hGH contains a convenient PstI site, CTGCAG. The plasmid p04 (SEQ ID NO: 9), a derivative of pUC19 (New England Biolabs) containing the same multi-cloning site as pET41a, is first readied by digesting with PstI, followed by Mung Bean Nuclease, and subsequent re-ligation to destroy the internal PstI site to create p04A1. Finally, the NdeI/BamHI hGH fragment from p0A0 is ligated into similarly digested p04A1 to yield p0A03.  
         [0170]    Several examples are now given to generate assemblies for GH multimers with different linkers. Variation in the linker sequence, as well as the degree of monomer polymerization, may alter the polymers ease of production, conformation, in vitro activity, in vivo activity, immunogenicity, etc.  
       Example 2  
       [0171]    The second example involves generation of an assembly for the direct fusion multimer of GH.  
         [0172]    There is not a convenient restriction pair at the termini of rhGH, so this example uses the methods for a monomer sequence with an internal restriction pair. A direct fusion assembly for hGH is constructed with the features diagrammed in FIG. 2. Disclosed are two 5′-terminal cassettes, the amplification cassettes, and two 3′-terminal cassettes. The 3′-terminal cassette is engineered to enable construction of an insertion cassette, as shown in FIG. 5. This facilitates insertion of amplification cassettes to generate expressible genes for different size homopolymeric GHs.  
         [0173]    Two 5′-terminal cassettes for the GH fusion protein assembly are disclosed. The first is a direct start 5′-terminal cassette, and the second is an OmpA start 5′-terminal cassette. The direct start results in an N-terminal methionine at the N-terminus of the final expressed GH polymer. Its construction is straight forward because the insert in p0A0 and p0A03 already has the N-terminal methionine fused to the GH gene. In contrast, the OmpA start codes for an N-terminal leader sequence that targets the polymer to the periplasmic space of  E. Coli , resulting in the cleavage of the leader from the polymer. There are many other 5′-terminal cassettes that can easily be generated by those skilled in the art.  
         [0174]    A pre-5′-terminal cassette is disclosed that enables fusion of the OmpA sequence to any other blunt end or HindIII digested sequence. SEQ ID NO: 10 is a synthetic DNA fragment that contains the coding sequence for the OmpA leader peptide, and its ORF translation is listed in SEQ ID NO: 11. The fragment has a 5′ NdeI site, the OmpA leader coding region, a 3′ HindIII site for HindIII ligation or blunt end ligation after filling in the HindIII 5′ overhang with T4 DNA polymerase, and a BamHI site for cloning flexibility. Plasmid p04 is readied by digestion to destroy an internal site, this time the HindIII site. The plasmid is digested with HindIII, followed by Mung Bean Nuclease, and subsequently ligated back together to create p04A2. Both p04A2 and insert DNA are digested with NdeI and BamHI and ligated together to yield the plasmid p0C0A2 as shown in FIG. 9.  
         [0175]    For the current use, a GH sequence is needed that contains a 5′ blunt end or HindIII site, along with a 3′ restriction site that is the 3′ member of a restriction pair. The 5′ terminus is engineered with a HindIII site. Digestion with Mung Bean Nuclease after digestion with HindIII results in a blunt 5′ end that leaves the 5′-terminal codon of GH, TTC, intact. Although the blunt end is not needed for the current example, in general it is necessary for ligation to other hypothetical cassettes.  
         [0176]    There are several choices for the restriction site pair, and we choose to use GH amino acids 187 and 188, glycine and serine, that are compatible with, among other enzymes, BamHI and BclI. The two enzymes recognize sequences GGATCC and TGATCA, respectively. BamHI is assigned as the 3′ member, and BclI is assigned as the 5′ member.  
         [0177]    The desired DNA sequence is generated by PCR using p0A03 as template, as shown in FIG. 10. The 5′ and 3′ primers are listed in SEQ ID NO: 12 and SEQ ID NO: 13, respectively, and the DNA coding region for the insert between the 5′ flanking NdeI and 3′ BamHI sites is listed in SEQ ID NO: 14. The fragment is digested with HindIII and BamHI and inserted into similarly digested p04B1 to yield p0A01. Plasmid p04B1 is prepared by destroying the HindIII site in p04A1 as described for the preparation of p04A2. The result is a parent plasmid with the PstI and HindIII sites destroyed.  
         [0178]    The 5′-terminal cassettes are now constructed from the generated sequences as shown in FIG. 10. The XbaI/HindIII fragment from p0C0A2 is inserted into plasmid p0A01 to generate p0A11A2. The result is the OmpA 5′-terminal cassette for the GH direct fusion assembly. It contains the OmpA sequence fused directly to the 5′ coding region of GH. The resulting DNA insert between NdeI and BamHI is listed in SEQ ID NO: 15, with corresponding ORF listed in SEQ ID NO: 16. The direct translation start 5′-terminal cassette is constructed by ligating fragments from existing sequences. The PstI/BamHI 5′ GH fragment and plasmid backbone that results from digesting p0A03 is ligated with the PstI/BamHI 3′ GH fragment that results from digesting p0A01 to yield p0A11A1. The resulting DNA sequence between NdeI and BamHI, and the corresponding ORF, for p0A11A1 are listed in SEQ ID NO: 17 and SEQ ID NO: 18, respectively.  
         [0179]    As shown in FIG. 2, the amplification cassette must contain several components. First, it must have both the 5′ and 3′ members of the restriction pair to enable polymerization. In between must be the entire continuous GH sequence. Finally, if convenient, there should be flanking restriction sites for insertion and extraction of the sequence from a plasmid backbone.  
         [0180]    The amplification cassette for the current direct fusion of GH is generated by PCR, as shown in FIG. 11. The 5′ primer is listed in SEQ ID NO: 19. It contains an NdeI site, the 5′ restriction pair member BclI, followed by the codons that together code for GH amino acids 187-191, and finally codons to anneal to the GH 5′-terminal codons. The 3′ primer is one previously used and listed in SEQ ID NO: 13. The PCR template is p0A03. The resulting insert DNA sequence between NdeI and BamHI is listed in SEQ ID NO: 20, with ORF sequence listed in SEQ ID NO: 21. The DNA sequence is inserted into plasmid p04A1 to yield p0A11B.  
         [0181]    Two simple 3′-terminal cassettes are disclosed, as shown in FIG. 12. Both code for the 3′ terminus of GH, starting at the glycine and serine codons within the BclI site, amino acids 187 and 188, and ending with the translation stop codon, TAG. The first cassette, given in SEQ ID NO. 22, is a direct translation stop. The double stranded DNA is synthesized and contains an EcoRI site flanking the 5′ terminus, a BclI site to ligate to BamHI, the 3′ terminus of GH, a stop codon, and a SalI site for cloning flexibility. It is inserted into p04A1 by digesting the synthetic DNA and p04A1 with EcoRI and SalI and ligating the large fragments together to yield plasmid p0A11C1. The C-terminal ORF protein sequence contributed by this cassette to subsequent GH multimer constructs is given in SEQ ID NO: 23.  
         [0182]    The second 3′-terminal cassette, given in SEQ ID NO: 24, is a synthetic DNA fragment similar to the first, except it contains the codons for a 3 amino acid polylysine tail before the stop codon. It is analogously inserted into p04A1 to yield plasmid p0A11C2. The polylysine tail is potentially useful for chemical conjugation with other molecules. SEQ ID NO: 25 is the C-terminal ORF sequence contributed by the new insert to subsequent GH multimer constructs.  
         [0183]    Once the basic cassettes are complete, the amplification cassette can be polymerized, the 5′-terminal and 3′-terminal cassettes can be joined to form an insertion cassette, and finally amplification cassettes can be ligated to the insertion cassette to generate expressible multimers.  
       Example 3  
       [0184]    The polymerization of the GH direct fusion amplification cassettes is performed as shown in general in FIG. 3 and specifically in FIG. 13. The first polymerization is formation of the dimer. Plasmid p0A11B is digested with NdeI and BclI and the plasmid isolated. In a separate reaction, p0A11B is digested with NdeI/BamHI and the insert isolated. The two fragments are then ligated together to yield plasmid p0A11B2. Its insert DNA sequence is listed in SEQ ID NO: 26, and the corresponding ORF translation is listed in SEQ ID NO: 27. This process is repeated, changing the identity, and thus the size, of amplification cassettes 1 and 2 in FIG. 13 to construct polymer inserts of different sizes. The size of new constructs is increased fastest if the polymerization is done geometrically, each time using the most recent construct for both cassettes 1 and 2. The size is increased by one if the monomer amplification cassette, p0A11B, is used either as cassette 1 or 2. The generalized sequences for the resulting amplification cassettes are given in SEQ ID NO: 28 and SEQ ID NO: 29 for the DNA and protein, respectively.  
       Example 4  
       [0185]    The cassettes for the GH direct fusion assembly are designed to enable construction of insertion cassettes to facilitate generation of a variety of expressible polymers. The general procedures are shown in FIGS. 5 and 7 and the specifics in FIG. 14. Different insertion cassettes can be generated with the various 5′-terminal and 3′-terminal cassettes. However, only the one involving p0A11A1 and p0A11C1 is described here. Others are constructed in exactly the same way.  
         [0186]    Plasmid p0A11A1 is digested with EcoRI and SalI and the opened plasmid is isolated. Plasmid p0A11C1 is digested with the same enzyme pair and the insert isolated. The two fragments are ligated together to generate the insertion cassette, p0A11D, and the resulting DNA sequence is listed in SEQ ID NO: 30. Plasmid p0A11D is compatible with ligation of any of the amplification cassettes for this assembly. It need be prepared only once for all subsequent ligations, as long as the supply is sufficient.  
       Example 5  
       [0187]    Either of the two schemes shown in FIGS.  6  and 7 can be used to ligate amplification cassettes into the insertion cassette. The example given here utilizes the oriented ligation shown in FIG. 7 and subsequent digestion and re-ligation to generate final products as shown in FIG. 14.  
         [0188]    Plasmid p0A11D is digested with BamHI and EcoRI, and the plasmid is isolated. An amplification cassette is digested with BclI and EcoRI and the insert isolated. Ligation of the two fragments yields an intermediate that is converted to the multimer expression cassette after digestion with BamHI and BclI, purification, and subsequent re-ligation. The result is an expression ready insert for the direct fusion growth hormone multimer. When performed with the Nmer amplification cassette, the result is an N+1 multimer expression cassette. The insert has general DNA sequence listed in SEQ ID NO: 31 and corresponding ORF translation listed in SEQ ID NO: 32. The production of different size multimers is controlled by the size of the ligated amplification cassette.  
         [0189]    Protein expression is achieved by digesting and ligating the multimer expression cassette insert into an appropriate expression system. For example, the insert can be liberated with NdeI and SalI and ligated into similarly digested pET41a, followed by transformation into  E. coli  strain BL21(DE3) (Novagen).  
         [0190]    One utility of the invention is the ease of production of different size multimers and different variations once the basic cassettes, p0A11A1, p0A11A2, p0A11B, p0A11C1, and p0A11C2, for example, are constructed. Those skilled in the art can easily see how substituting p0A11C2 for p0A11C1 when generating the insertion cassette generates a polylysine tail variant.  
       Example 6  
       [0191]    The next example involves generation of a GH multimer with a linker without a convenient restriction pair. The one amino acid linker, glycine, is used as an example. The construction of GH multimers with a glycine linker is analogous to the construction of the fusion protein. In fact, the GH glycine linker assembly shares the same 5′- and 3′-terminal cassettes with the GH fusion protein assembly. This is one advantage of the assembly construction scheme given in FIG. 2. Assemblies differing only in the linker region only need different amplification cassettes, while sharing the same 5′- and 3′-terminal cassettes.  
         [0192]    Use p0A11A1 and p0A11A2 as before for the direct start and OmpA 5′-terminal cassettes for the direct fusion assembly. Use p0A11C1 and p0A11C2 as before for direct stop and poly lysine 3′-terminal cassettes.  
         [0193]    The only difference is the amplification cassettes that contain a glycine codon between the ending and starting codons for GH. The glycine linker amplification cassette is made in the same way as the one for the direct fusion homomultimer except for some necessary substitutions of sequences, as shown in FIG. 15. SEQ ID NO: 33 is substituted for SEQ ID NO: 19 as the 5′ PCR primer. It contains the same elements as before, as well as the glycine codon between the sequence for amino acids 191 and 1. The resulting PCR fragment is inserted into parent plasmid p04A1 by digesting both the parent plasmid and the PCR fragment with NdeI and BamHI and ligating the appropriate fragments together. The resulting plasmid is labeled p0A21B. The DNA sequence and ORF translation for the insert sequence between NdeI and BamHI are listed in SEQ ID NO: 34 and SEQ ID NO: 35, respectively.  
         [0194]    The construction of additional amplification assemblies, the insertion cassette, and multimer expression cassettes for the GH glycine linker assembly is identical in practice to the one for the GH direct fusion assembly, FIGS. 13 and 14, except for the substitution of p0A21B for p0A11B. The corresponding generalized amplification cassette insert DNA and ORF sequences are listed in SEQ ID NO: 36 and SEQ ID NO: 37, and the general formulas for the multimer expression cassettes are listed in SEQ ID NO: 38 and SEQ ID NO: 39.  
         [0195]    The previous examples have demonstrated, among other things, the ease at which multiple 5′- and 3′-terminal cassettes can be used to introduce variations in the N- and C-termini of a polymer. In the case of the 5′-terminal cassettes, cassettes with either a direct translation start or one introducing a leader sequence are disclosed. In the case of the 3′-terminal cassettes, ones with either a direct stop or one introducing a polylysine tail are disclosed. Each demonstrates the ease at which functional elements can be added to the beginning or end of a polymer sequence. These methods are easily extended to other examples by those skilled in the art. Therefore, subsequent examples will be limited to the presentation of only a single 5′- and 3′-terminal cassette for each assembly.  
         [0196]    The next examples involve generation of GH multimers utilizing linkers that result in monomers with a terminal restriction pair. FIG. 1 details the general features for these assemblies.  
       Example 7  
       [0197]    This example involves a linker that is noteworthy because it contains a 3′ restriction pair member with a functional stop codon that is destroyed upon polymerization. Use of this linker makes it possible to express functional multimers using just the 5′-terminal and amplification cassettes. However, a 3′-terminal cassette is necessary to express homomultimers without any residual linker at the 3′ terminus of the protein.  
         [0198]    The 5′ restriction pair member is NcoI, C{circumflex over ( )}CATGG, while the 3′ restriction pair member is RcaI, T{circumflex over ( )}CATGA. Therefore, the resulting linker sequence is A-Ser-Trp-B, where A and B are arbitrary protein sequences. For the given example, A is a null sequence, and B is G 4 S, where the single letter amino acid abbreviations are used.  
         [0199]    For this example, only one 5′-terminal cassette is disclosed, with a direct ATG start codon and no leader sequence, as shown in FIG. 16. The PCR primers for the 5′-terminal cassette are listed in SEQ ID NO: 3 and SEQ ID NO: 40, for the 5′ and 3′ ends, respectively. The 5′ primer maintains the NdeI site and its start codon, while the 3′ primer introduces a stop codon within an RcaI (or BspHI) restriction site, immediately followed by a BamHI site. The template for the reaction is p0A0.  
         [0200]    Because the RcaI restriction site also contains the codon TCA immediately 5′ of the stop codon, it also introduces a C-terminal serine residue. The resulting PCR fragment is purified and ligated into pET41a in an analogous manner for the generation of p0A0. The sequence verified plasmid is labeled p0A31A, and the DNA coding region, from the NdeI to the BamHI site, and the resulting ORF protein sequence are listed in SEQ ID NO: 41 and SEQ ID NO: 42, respectively. Expression of protein from the gene for p0A31A yields a 193 amino acid protein consisting of full length hGH with an additional N-terminal methionine and C-terminal serine.  
         [0201]    The PCR primers for the amplification cassette are listed in SEQ ID NO: 43 and SEQ ID NO: 40, for the 5′ and 3′ ends, respectively. The 5′ primer introduces an NcoI site followed by the linker region. The NcoI site is ligation compatible with the 3′ RcaI site, and any such ligation destroys the TGA stop codon by altering it to a TGG codon. The resulting PCR fragment is purified and ligated into pET41a after the PCR product and plasmid are cut with NcoI and BamHI, as shown in FIG. 16. The sequence verified plasmid is labeled p0A31B, and the DNA coding region from the NcoI to the BamHI site is listed in SEQ ID NO: 44. The ORF protein sequence coded by the insert is given in SEQ ID NO: 45.  
         [0202]    Again, for this example, only one 3′-terminal cassette is disclosed, with a direct TAG-stop codon and no other 3′-specific sequences. The 3′-terminal cassette is constructed using PCR with p0A0 as template and SEQ ID NO: 43 and SEQ ID NO: 4 as 5′ and 3′ primers, respectively. This creates a cassette with a 5′ linker and a 3′ stop codon immediately following the last amino acid from the parent monomer. The PCR fragment is inserted into pET41a as before and shown in FIG. 16 to create p0A31C. The resulting DNA and protein fragments between the NdeI and BamHI sites are listed in SEQ ID NO: 46 and SEQ ID NO: 47, respectively.  
         [0203]    The scheme for the polymerization of the amplification cassettes is shown in FIG. 3. Additional care is necessary because the parent plasmid contains RcaI sites. One way to unambiguously liberate the insert sequence for polymerization is to first digest the flanking BamHI site, isolate the insert, and then digest with RcaI. The general formulas for the Nmer amplification cassette are listed in SEQ ID NO: 48 and SEQ ID NO: 49 for the DNA and corresponding ORF translation, respectively.  
       Example 8  
       [0204]    The ligation of the multimer assembly cassettes must be done sequentially, as shown in FIG. 4, because the arrangement of the restriction sites in the 3′-terminal cassette is like FIG. 2 d . The first ligation involves the 5′-terminal and amplification cassettes, rather than the 3′-terminal and amplification cassettes, to take advantage of the stop codon in the 3′-restriction member to produce expression ready inserts. The specifics are shown in FIG. 17 using procedures already described. Use of the monomeric amplification cassette, p0A31B, results in the dimeric cassette, p0A31F2, with insert DNA and corresponding ORF translation listed in SEQ ID NO: 50 and SEQ ID NO: 51. The general formulas for the N+1 mer produced after ligation between the Nmer amplification and the 5′-terminal cassettes are listed in SEQ ID NO: 52 and SEQ ID NO: 53. Transfer of the insert into an appropriate expression system yields expression of the N+1 GH polymer with the SWG4S linker and C-terminal S residue.  
         [0205]    Completion of the ligation scheme shown in FIG. 17 results in an insert with an additional monomer and the natural C-terminus of GH. If the insert from p0A31F2 is ligated into p0A31C, then the trimer expression cassette p0A31E3 is generated. In general, the formulas for the insert DNA and corresponding ORF translation when the Nmer amplification cassette is used are listed in SEQ ID NO: 54 and SEQ ID NO: 55. For p0A31E3, the monomer amplification cassette is used and N=1.  
       Example 9  
       [0206]    The plasmids containing the inserts generated with the ligation scheme shown in FIG. 17 are capable of expressing rhGH polymers following standard techniques (see for example, user manuals from Novagen, Madison, Wis.). DNA sequences listed in SEQ ID NO: 52 with N=0, 1, 2, 4, and 8 and prepared according to Example 8 are ligated into pET41a. The resulting plasmids are separately transformed into BL21(DE3) and separately grown in Luria Browth medium and induced to express the polymer protein by adding IPTG to a concentration of 1 mM.  
         [0207]    Following 3 hours of induction, each culture is harvested by centrifugation and treated with SDS-PAGE sample buffer. Proteins from the samples for each culture are separated according to their molecular weights on a standard SDS-PAGE gel (Invitrogen, Carlsbad, Calif.). The resulting gel is stained with coomasie blue stain to visualize the protein bands Results for the monomer (SEQ ID NO: 42), dimer (N=1 in SEQ ID NO: 53), trimer (N=2 in SEQ ID NO: 53), pentamer (N=4 in SEQ ID NO: 53), and nanamer (N=8 in SEQ ID NO: 53) are given in FIG. 18. As the figure demonstrates, large amounts of each polymeric rhGH are produced except for the nanamer.  
       Example 10  
       [0208]    Linkers with convenient restriction sites offer the engineering option to generate a multitude of assemblies with cassettes that can be attached to monomers using restriction/ligation techniques. The utility of this formulation lies in the breadth of assemblies that can be constructed relatively easily. This is especially apparent when the linkers themselves are treated as assemblies nested within the construction of the multimers. Once constructed, these linker assemblies and cassettes, like any other, can be reused to produce new assemblies.  
         [0209]    Nested linker assemblies are constructed having a slightly different function than the multimer assemblies. They still need an amplification cassette for the polymerization of the linker. However, the other cassettes in the assembly enable attachment of the linker to either a 5′or 3′ terminus, whichever is appropriate.  
         [0210]    The example given here is a series of linkers, having amino acid sequence GZGS, where Z is an arbitrary sequence of arbitrary length. The series of linkers in Table 1 below share features that enable them to be treated similarly in terms of their engineering. All but one has a Glycine at the N-terminus of the linker that can be coded by an NaeI restriction site at the 5′ end for blunt end ligation of a 5′-terminal cassette to a monomer pre-cassette. For the other linker, GS, a synthetic DNA fragment must be ligated to the monomer pre-cassette without propagation within a plasmid. Each of the linkers ends in the protein sequence GS, so that the restriction pair is identical to earlier examples utilizing the BclI and BamHI sites.  
                                         TABLE 1                       Linker protein   5′-terminal cassette DNA   Amplification cassette DNA       monomer unit   sequence   sequence                                GS   GGATCC   TGATCAGGATCC                   GGS   GCCGGCGGATCC   TGATCAGGCGGATCC               GGGS   GCCGGCGGCGGATCC   TGATCAGGCGGCGGATCC               GGGGS   GCCGGCGGCGGCGGATCC   TGATCAGGCGGCGGCGGATCC               GZGS   GCCGGCYGGATCC   TGATCAGGCYGGATCC           Z is an arbitrary protein           sequence, and Y is its DNA coding           sequence.                  
 
         [0211]    As a single example of the engineering of the linker assembly, we construct the (G 4 S) x  linker, where x indicates the degree of polymerization of the monomer sequence. The assembly is engineered like any other, and it falls into the scheme shown in FIG. 1. The specifics are shown in FIG. 19. Two synthetic DNA sequences are needed, SEQ. ID NO: 56 and SEQ ID NO: 57.  
         [0212]    The first, the 5′-terminal cassette labeled as p0D11A in FIG. 19, is the sequence enabling addition of the linker sequence to other cassettes. It is flanked by a NcoI site, and thus with an upstream NdeI site, for cloning flexibility at the 5′ terminus, contains the NaeI site to create the blunt end ligation with the glycine codon at the 5′ terminus, the linker sequence, and finally the BamHI site within the GS codons. Plasmid p04 is prepared by digestion with NgoMIV, digestion with Mung Bean Nuclease, and finally re-ligation to destroy the internal NaeI site, creating plasmid p04A3. This altered plasmid, along with the insert, is digested with NcoI and BamHI and the appropriate fragments are ligated together. The resulting plasmid is labeled p0D11A. The open reading frame translation between the cleaved NaeI and the entire BamHI sites is G 4 S.  
         [0213]    SEQ ID NO: 57 is the sequence for the amplification cassette to create multimers of the G 4 S linker. It is flanked by an NcoI site, again for cloning flexibility. It has the 5′ BclI site from the restriction pair, followed by the G 4 S coding sequence that ends with the BamHI site. It is inserted into p04 by cutting both plasmid and insert with NcoI and BamHI and ligating the appropriate fragments together, as shown in FIG. 19. The resulting plasmid is labeled p0D11B.  
         [0214]    Amplification cassette p0D11B is polymerized by the scheme shown in FIG. 3, left hand side, to create a dimer. In this instance the decision to follow the left hand side scheme results in larger fragments that are easier to isolate. Plasmid p0D11B is digested with NdeI and BclI and the large fragment is isolated. Separately, the same parent plasmid is digested with NdeI and BamHI, this time isolating the small fragment. The two isolated fragments are then ligated together, destroying the internal BclI and BamHI sites, but preserving the flanking ones. The resulting plasmid is labeled p0D11B2, the DNA insert is listed in SEQ ID NO: 58, and the ORF translation is listed in SEQ ID NO: 59. The sequence codes for the dimer (G 4 S) 2 . The process can be repeated with different starting cassettes to generate any (G 4 S) x  linker. In this manner, (G 4 S) 4  can be generated by digesting p0D12B with NdeI and BclI and saving the large fragment and ligating in the small fragment generated by digesting it with NdeI and BamHI.  
         [0215]    The engineering of the G 4 S assembly enables the construction of a GH multimer assembly with the (G 4 S) 3  linker. The (G 4 S) 3  5′-terminal cassette for ligation to the GH sequences is generated following the general scheme shown in FIG. 4. Plasmid p0D11B2 is digested with NdeI and BclI, and the large fragment is isolated. The small fragment resulting from digestion of p0D11A with NdeI and BamHI is ligated in, creating plasmid p0D13A. The DNA and ORF sequences for the insert are listed in SEQ ID NO: 60 and SEQ ID NO: 61, respectively. The insert in p0D13A enables ligation of the (G 4 S) 3  linker to the 3′ end of any sequence ending in a blunt end.  
       Example 11  
       [0216]    Engineering of the GH (G 4 S) 3  assembly requires two new ends to the GH gene. The BclI 5′ restriction pair member is needed on the 5′ terminus of the amplification and 3′-terminal cassettes, and a blunt end immediately after the last codon of GH is needed on the 3′ terminus of the 5′-terminal and amplification cassettes for ligation of the (G 4 S) 3  linker. There are many ways to get a blunt end at the 3′ terminus of GH. Disclosed here is the use of an NcoI site that is made blunt after digestion with Mung Bean Nuclease. In addition, it is convenient to introduce a stop codon flanked by the SalI restriction site at the 3′ terminus of the GH gene for construction of an insertion cassette, as shown in general in FIG. 5.  
         [0217]    Three new primers are used to generate the new termini on two new GH inserts by PCR using P0A03 as template, as shown in FIG. 20. The 5′ primer is listed in SEQ ID NO: 62. It contains a flanking NdeI site, the BclI 5′ restriction pair member, and sequence complementary to the GH 5′ terminus. It is used for both PCR reactions. The 3′ primers are listed in SEQ ID NO: 63 and SEQ ID NO: 64. Both contain sequence complementary to the 3′ terminus of GH. The first codes for the NcoI site at the 3′ terminus for creation of a blunt end after the last GH base pair and a flanking EcoRI site, while the second introduces a stop codon followed by a SalI restriction site.  
         [0218]    The PCR fragments are ligated into plasmid backbones as shown in FIG. 20. The PCR fragment resulting from use of the primers listed in SEQ ID NO: 62 and SEQ ID NO: 63 is digested with NdeI and EcoRI and ligated into similarly digested p04A1 to yield p0A04, while the fragment resulting from use of the primers listed in SEQ ID NO: 62 and SEQ ID NO: 64 is digested with BclI and SalI and ligated into similarly digested p0A11C1 to give p0A41C. The insert in p0A04 between the BclI and blunt ended NcoI sites has the DNA sequence listed in SEQ ID NO: 65 and corresponding ORF translation listed in SEQ ID NO: 66. Likewise, the insert in p0A41C, the 3′-terminal cassette, between the BclI and SalI sites has the DNA sequence listed in SEQ ID NO: 67 and ORF translation listed in SEQ ID NO: 68.  
         [0219]    The amplification cassette is generated first by ligating the (G 4 S) 3  linker from plasmid p0D13A with the insert in p0A04, as shown in FIG. 21. Plasmid p0D13A is digested with NaeI and HindIII, and the small fragment is isolated. It is ligated into p0A04 after digestion first with NcoI, then Mung Bean Nuclease, and finally HindIII to yield p0A43B. The resulting DNA sequence for the amplification cassette between the BclI and BamHI sites is listed in SEQ ID NO: 69, with corresponding ORF translation in SEQ ID NO: 70.  
         [0220]    The direct start 5′-terminal cassette is generated by combining the 5′ elements from p0A11A1 with the 3′ elements from p0A43B, as shown in FIG. 21. The small fragment resulting from digesting p0A43B with PstI and EcoRI is isolated. It is ligated to the large fragment resulting from digestion of p0A11A1 with the same enzymes to yield p0A43A. The DNA sequence for the insert between NdeI and BamHI is listed in SEQ ID NO: 71, with corresponding ORF translation in SEQ ID NO: 72.  
         [0221]    The polymerization of the amplification cassettes again follows the scheme in FIG. 3. The general formulas for the insert DNA and corresponding ORF translation for the Nmer amplification cassette are listed in SEQ ID NO: 73 and SEQ ID NO: 74.  
         [0222]    The ligation of the cassettes for the GH (G 4 S) 3  linker assembly to create a multimer expression cassette follows the previously described scheme shown in FIG. 7 and demonstrated in Example 4. The insertion cassette is first generated with the 5′- and 3′-terminal cassettes using EcoRI and SalI digestions. An amplification cassette insert is first isolated after digestion with BclI and EcoRI and then spliced into the insertion cassette after digestion using BamHI and EcoRI. The resultant construct is subsequently digested with BamHI and BclI and re-ligated. The resulting N+2 multimer expression cassette, where N is the degree of polymerization of the amplification cassette used, has DNA and corresponding ORF translation sequences listed in SEQ ID NO: 75 and SEQ ID NO: 76. Transfer of the insert into a suitable expression system yields multimeric GH with (G 4 S) 3  linker.  
       Example 12  
       [0223]    The last example is an alternative construction for a GH direct fusion assembly. It involves the use of an incompatible restriction pair that is blunt ended for ligation. Construction of this new assembly is done by ligating together fragments from earlier cassettes, since they already contain the needed elements. The construction scheme is shown in FIG. 22.  
         [0224]    The 5′-terminal cassette is labeled p0A51A. It is generated by combining elements from p0A11A1 and p0A04. Plasmid p0A11A1 is digested with PstI and EcoRI and the open plasmid isolated. This is ligated with the insert isolated after digesting p0A04 with the same enzymes. The result, p0A51A, has DNA and corresponding ORF translation listed in SEQ ID NO: 77 and SEQ ID NO: 78.  
         [0225]    The amplification and 3′-terminal cassettes are constructed in exactly the same manner as the 5′-terminal cassette, except for substituting which plasmids are digested. For the amplification cassette, plasmid p0A01 is ligated with the insert from p0A04. The insert DNA and corresponding ORF sequences are listed in SEQ ID NO: 79 and SEQ ID NO. 80. Likewise, for the 3′-terminal cassette, plasmid p0A01 is ligated with the insert from p0A03. Its insert DNA and corresponding ORF translation are listed in SEQ ID NO: 81 and SEQ ID NO: 82.  
         [0226]    The polymerization of amplification cassettes still follows the scheme in FIG. 3. However, digestion at a restriction pair member now requires the additional blunt ending of its overhang. FIG. 23 shows the specifics for the current assembly. The digestions of the cassette are done sequentially so that the restriction pair is blunt ended, but the flanking restriction sites are left intact. The general formulas for the amplification cassettes are listed in SEQ ID NO: 83 and SEQ ID NO: 84.  
         [0227]    The ligation of the multimer assembly cassettes is done sequentially as shown in FIG. 4. The digestion of any plasmid is performed as described above with blunt ending of the restriction pair member first. The general formulas for the resulting multimer expression cassette insert, using the Nmer amplification cassette, are listed in SEQ ID NO: 85 and SEQ ID NO: 86.  
         [0228]    In practice, ligations of cassettes from this assembly involves more steps, but the technique&#39;s almost universal applicability may make it the method of choice in some instances. For the current case, the assembly given in Examples 1-4 is easier to manipulate.  
         [0229]    Those skilled in the art will recognize many equivalents to the examples presented herein, using different monomers, linkers, restriction pairs, flanking restriction sites, 5′ specific sequences, 3′ specific sequences, and ligation strategies. For example, the methods are flexible as to the order of ligating 5′-terminal cassettes, 3′-terminal cassettes, and amplification cassettes, and in ligating amplification cassettes to one another to form higher order amplification cassettes. Combining elements of the following claims presented here and in the description, including the examples, is within the scope of the invention and are encompassed in the following claims.  
         [0230]    All references cited herein, including the bibliography, are incorporated by reference in their entireties.  
       References Cited  
     U.S. PATENT DOCUMENTS  
       [0231]    [0231]                                                           5,084,390   January 1992 Hallewell et al.   435/188           5,876,969   March 1999 Fleer et al.   435/69.7           6,242,570   June 2001 Sytkowski   530/350                        
       OTHER PUBLICATIONS  
       [0232]    Jorgensen, J. O. L., “Human Growth Hormone Replacement Therapy: Pharmacological and Clinical Aspects,”  Endocrine Reviews , 12(3):189, (1991).  
         [0233]    Chien, Y. W., “Human Insulin: Basic Sciences to Therapeutic Uses,”  Drug Development and Industrial Pharmacy , 22(8):753-789 (1996).  
         [0234]    Putney, S. D., and P. Burke, “Improving Protein Therapeutics with Sustained-release Formulations,”  Nature Biotechnology , 16:153, (1998).  
         [0235]    Johnson, O. L., et al., “A Month-long Effect from a Single Injection of Microencapsulated Human Growth Hormone,”  Nature Medicine , 2(7):795, (1996).  
         [0236]    Chen, S. A., et al., “Plasma and Lymph Pharmacokinetics of Recombinant Human Interleukin-2 and Polyethylene Glycol-modified Interleukin-2 in Pigs,”  The Journal of Pharmacology and Experimental Therapeutics , 293(1):248, (2000).  
         [0237]    Venkatachalam, M. A., and H. Rennke, “The Structural and Molecular Basis of Glomerular Filtration,”  Circulation Research , 43(3):337-347, (1978).  
         [0238]    Roberts, M. J., et al., “Chemistry for Peptide and Protein PEGylation,”  Adv Drug Deliv Rev , 54(14):459-76, (2002).  
         [0239]    Sharieff, K. A., et al., “Advances in Treatment of Chronic Hepatitis C: ‘Pegylated’ Interferons,”  Cleve Clin J Med , 69(2):155-9, (2002).  
         [0240]    Lewis, R. V., et al., “Expression and Purification of a Spider Silk Protein: A New Strategy for Producing Repetitive Proteins,”  Protein Expression and Purification , 7:400-6, (1996).  
         [0241]    Elmorjani, K., et al., “Synthetic Genes Specifying Periodic Polymers Modeled on the Repetitive Domain of Wheat Gliadins: Conception and Expression,”  Biochemical and Biophysical Research Communication , 239:240-6, (1997).  
         [0242]    Pennell, C. A., and P. Eldin, “In vitro Production of Recombinant Antibody Fragments in  Pichia pastoris,” Res Immunol , 149(6):599-603, (1998).  
         [0243]    Sambrook, et al., “Molecular Cloning: A Laboratory Manual,” Cold Spring Harbor Laboratory Press, NY, (2000).  
         [0244]    Aggarwal, B. B. and Gutterman, J. U. Eds., “Human Cytokines: Handbook for Basic and Clinical Research,” Blackwell Scientific Publications, Boston, Mass., (1992).  
     
       
       
         1 
         
           
             86  
           
           
             1  
             573  
             DNA  
             Homo sapiens  
           
            1 

ttcccaacca ttcccttatc caggcttttt gacaacgcta tgctccgcgc ccatcgtctg     60 

caccagctgg cctttgacac ctaccaggag tttgaagaag cctatatccc aaaggaacag    120 

aagtattcat tcctgcagaa cccccagacc tccctctgtt tctcagagtc tattccgaca    180 

ccctccaaca gggaggaaac acaacagaaa tccaacctag agctgctccg catctccctg    240 

ctgctcatcc agtcgtggct ggagcccgtg cagttcctca ggagtgtctt cgccaacagc    300 

ctggtgtacg gcgcctctga cagcaacgtc tatgacctcc taaaggacct agaggaaggc    360 

atccaaacgc tgatggggag gctggaagat ggcagccccc ggactgggca gatcttcaag    420 

cagacctaca gcaagttcga cacaaactca cacaacgatg acgcactact caagaactac    480 

gggctgctct actgcttcag gaaggacatg gacaaggtcg agacattcct gcgcatcgtg    540 

cagtgccgct ctgtggaggg cagctgtggc ttc                                 573 

 
           
             2  
             191  
             PRT  
             Homo sapiens  
             
               mat_peptide  
               (1)..()  
             
           
            2 

Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg 
1               5                   10                  15 

Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu 
            20                  25                  30 

Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro 
        35                  40                  45 

Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg 
    50                  55                  60 

Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu 
65                  70                  75                  80 

Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val 
                85                  90                  95 

Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp 
            100                 105                 110 

Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu 
        115                 120                 125 

Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser 
    130                 135                 140 

Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr 
145                 150                 155                 160 

Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe 
                165                 170                 175 

Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

 
           
             3  
             38  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            3 

ggaattccat atgttcccaa ccattccctt atccaggc                             38 

 
           
             4  
             36  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            4 

cgcggatccc tagaagccac agctgccctc cacaga                               36 

 
           
             5  
             19  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            5 

taatacgact cactatagg                                                  19 

 
           
             6  
             21  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            6 

tgctagttat tgctcagcgg t                                               21 

 
           
             7  
             588  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            7 

catatgttcc caaccattcc cttatccagg ctttttgaca acgctatgct ccgcgcccat     60 

cgtctgcacc agctggcctt tgacacctac caggagtttg aagaagccta tatcccaaag    120 

gaacagaagt attcattcct gcagaacccc cagacctccc tctgtttctc agagtctatt    180 

ccgacaccct ccaacaggga ggaaacacaa cagaaatcca acctagagct gctccgcatc    240 

tccctgctgc tcatccagtc gtggctggag cccgtgcagt tcctcaggag tgtcttcgcc    300 

aacagcctgg tgtacggcgc ctctgacagc aacgtctatg acctcctaaa ggacctagag    360 

gaaggcatcc aaacgctgat ggggaggctg gaagatggca gcccccggac tgggcagatc    420 

ttcaagcaga cctacagcaa gttcgacaca aactcacaca acgatgacgc actactcaag    480 

aactacgggc tgctctactg cttcaggaag gacatggaca aggtcgagac attcctgcgc    540 

atcgtgcagt gccgctctgt ggagggcagc tgtggcttct agggatcc                 588 

 
           
             8  
             192  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            8 

Met Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

 
           
             9  
             2907  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            9 

ccggatatag ttcctccttt cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg     60 

ggttatgcta gttattgctc agcggtggca gcagccaact cagcttcctt tcgggctttg    120 

tttagcagcc taggtattaa tcaattagtg gtggtggtgg tggtggtggt gctcgagtgc    180 

ggccgcaagc ttgtcgacgg agctcgcctg caggcgcgcc aaggcctgta cagaattcgg    240 

atccccgata tcccatggga ctcttgtcgt cgtcatcacc ggagccacca ccggtaccca    300 

gatctgggct gtccatgtgc tggcgttcga atttagcagc agcggtttct ttcataccaa    360 

ttgcagtact accgcgtggc accagacccg cggagtgatg gtgatggtga tgaccagaac    420 

cactagtaca cacatatgta tatctccttc ttaaagttaa acaaaattat ttctagaggg    480 

gaattgttat ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga    540 

tcgatctcga tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg    600 

gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg    660 

ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg    720 

ggcgccatct ccttgcatgc atggcgtaat catggtcata gctgtttcct gtgtgaaatt    780 

gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg    840 

gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt    900 

cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt    960 

tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc   1020 

tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg   1080 

ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg   1140 

ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac   1200 

gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg   1260 

gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct   1320 

ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg   1380 

tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct   1440 

gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac   1500 

tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt   1560 

tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc   1620 

tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca   1680 

ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat   1740 

ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac   1800 

gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt   1860 

aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc   1920 

aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg   1980 

cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg   2040 

ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc   2100 

cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta   2160 

ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg   2220 

ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct   2280 

ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta   2340 

gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg   2400 

ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga   2460 

ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt   2520 

gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca   2580 

ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt   2640 

cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt   2700 

ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga   2760 

aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt   2820 

gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc   2880 

gcacatttcc ccgaaaagtg ccacctg                                       2907 

 
           
             10  
             73  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            10 

cgccatatga aaaagacagc tatcgcgatt gcagtggcac tggctggttt cgctaccgta     60 

gcgcaagctt gag                                                        73 

 
           
             11  
             21  
             PRT  
             Escherichia coli  
             
               SIGNAL  
               (1)..(21)  
             
           
            11 

Met Lys Lys Thr Ala Ile Ala Ile Ala Val Ala Leu Ala Gly Phe Ala 
1               5                   10                  15 

Thr Val Ala Gln Ala 
            20 

 
           
             12  
             42  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            12 

ggacatatgc tgaagctttc ccaaccattc ccttatccag gc                        42 

 
           
             13  
             28  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            13 

cgcggatccc tccacagagc ggcactgc                                        28 

 
           
             14  
             578  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            14 

catatgctga agctttccca accattccct tatccaggct ttttgacaac gctatgctcc     60 

gcgcccatcg tctgcaccag ctggcctttg acacctacca ggagtttgaa gaagcctata    120 

tcccaaagga acagaagtat tcattcctgc agaaccccca gacctccctc tgtttctcag    180 

agtctattcc gacaccctcc aacagggagg aaacacaaca gaaatccaac ctagagctgc    240 

tccgcatctc cctgctgctc atccagtcgt ggctggagcc cgtgcagttc ctcaggagtg    300 

tcttcgccaa cagcctggtg tacggcgcct ctgacagcaa cgtctatgac ctcctaaagg    360 

acctagagga aggcatccaa acgctgatgg ggaggctgga agatggcagc ccccggactg    420 

ggcagatctt caagcagacc tacagcaagt tcgacacaaa ctcacacaac gatgacgcac    480 

tactcaagaa ctacgggctg ctctactgct tcaggaagga catggacaag gtcgagacat    540 

tcctgcgcat cgtgcagtgc cgctctgtgg agggatcc                            578 

 
           
             15  
             630  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            15 

catatgaaaa agacagctat cgcgattgca gtggcactgg ctggtttcgc taccgtagcg     60 

caagctttcc caaccattcc cttatccagg ctttttgaca acgctatgct ccgcgcccat    120 

cgtctgcacc agctggcctt tgacacctac caggagtttg aagaagccta tatcccaaag    180 

gaacagaagt attcattcct gcagaacccc cagacctccc tctgtttctc agagtctatt    240 

ccgacaccct ccaacaggga ggaaacacaa cagaaatcca acctagagct gctccgcatc    300 

tccctgctgc tcatccagtc gtggctggag cccgtgcagt tcctcaggag tgtcttcgcc    360 

aacagcctgg tgtacggcgc ctctgacagc aacgtctatg acctcctaaa ggacctagag    420 

gaaggcatcc aaacgctgat ggggaggctg gaagatggca gcccccggac tgggcagatc    480 

ttcaagcaga cctacagcaa gttcgacaca aactcacaca acgatgacgc actactcaag    540 

aactacgggc tgctctactg cttcaggaag gacatggaca aggtcgagac attcctgcgc    600 

atcgtgcagt gccgctctgt ggagggatcc                                     630 

 
           
             16  
             208  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            16 

Met Lys Lys Thr Ala Ile Ala Ile Ala Val Ala Leu Ala Gly Phe Ala 
1               5                   10                  15 

Thr Val Ala Gln Ala Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp 
            20                  25                  30 

Asn Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr 
        35                  40                  45 

Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser 
    50                  55                  60 

Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro 
65                  70                  75                  80 

Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu 
                85                  90                  95 

Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln 
            100                 105                 110 

Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp 
        115                 120                 125 

Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr 
    130                 135                 140 

Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe 
145                 150                 155                 160 

Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala 
                165                 170                 175 

Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp 
            180                 185                 190 

Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly 
        195                 200                 205 

 
           
             17  
             570  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            17 

catatgttcc caaccattcc cttatccagg ctttttgaca acgctatgct ccgcgcccat     60 

cgtctgcacc agctggcctt tgacacctac caggagtttg aagaagccta tatcccaaag    120 

gaacagaagt attcattcct gcagaacccc cagacctccc tctgtttctc agagtctatt    180 

ccgacaccct ccaacaggga ggaaacacaa cagaaatcca acctagagct gctccgcatc    240 

tccctgctgc tcatccagtc gtggctggag cccgtgcagt tcctcaggag tgtcttcgcc    300 

aacagcctgg tgtacggcgc ctctgacagc aacgtctatg acctcctaaa ggacctagag    360 

gaaggcatcc aaacgctgat ggggaggctg gaagatggca gcccccggac tgggcagatc    420 

ttcaagcaga cctacagcaa gttcgacaca aactcacaca acgatgacgc actactcaag    480 

aactacgggc tgctctactg cttcaggaag gacatggaca aggtcgagac attcctgcgc    540 

atcgtgcagt gccgctctgt ggagggatcc                                     570 

 
           
             18  
             188  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            18 

Met Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly 
            180                 185 

 
           
             19  
             52  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            19 

taccatatga catgatcatg tggcttcttc ccaaccattc ccttatccag gc             52 

 
           
             20  
             588  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            20 

catatgacat gatcatgtgg cttcttccca accattccct tatccaggct ttttgacaac     60 

gctatgctcc gcgcccatcg tctgcaccag ctggcctttg acacctacca ggagtttgaa    120 

gaagcctata tcccaaagga acagaagtat tcattcctgc agaaccccca gacctccctc    180 

tgtttctcag agtctattcc gacaccctcc aacagggagg aaacacaaca gaaatccaac    240 

ctagagctgc tccgcatctc cctgctgctc atccagtcgt ggctggagcc cgtgcagttc    300 

ctcaggagtg tcttcgccaa cagcctggtg tacggcgcct ctgacagcaa cgtctatgac    360 

ctcctaaagg acctagagga aggcatccaa acgctgatgg ggaggctgga agatggcagc    420 

ccccggactg ggcagatctt caagcagacc tacagcaagt tcgacacaaa ctcacacaac    480 

gatgacgcac tactcaagaa ctacgggctg ctctactgct tcaggaagga catggacaag    540 

gtcgagacat tcctgcgcat cgtgcagtgc cgctctgtgg agggatcc                 588 

 
           
             21  
             191  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            21 

Ser Cys Gly Phe Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn 
1               5                   10                  15 

Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr 
            20                  25                  30 

Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe 
        35                  40                  45 

Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr 
    50                  55                  60 

Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu 
65                  70                  75                  80 

Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe 
                85                  90                  95 

Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser 
            100                 105                 110 

Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu 
        115                 120                 125 

Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys 
    130                 135                 140 

Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu 
145                 150                 155                 160 

Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys 
                165                 170                 175 

Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly 
            180                 185                 190 

 
           
             22  
             42  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            22 

tacgaattcc attgatcatg tggcttctag taggtcgacg at                        42 

 
           
             23  
             4  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            23 

Ser Cys Gly Phe 
1 

 
           
             24  
             51  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            24 

tacgaattcc attgatcatg tggcttcaaa aagaaatagt aggtcgacga t              51 

 
           
             25  
             7  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            25 

Ser Cys Gly Phe Lys Lys Lys 
1               5 

 
           
             26  
             1161  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            26 

catatgacat gatcatgtgg cttcttccca accattccct tatccaggct ttttgacaac     60 

gctatgctcc gcgcccatcg tctgcaccag ctggcctttg acacctacca ggagtttgaa    120 

gaagcctata tcccaaagga acagaagtat tcattcctgc agaaccccca gacctccctc    180 

tgtttctcag agtctattcc gacaccctcc aacagggagg aaacacaaca gaaatccaac    240 

ctagagctgc tccgcatctc cctgctgctc atccagtcgt ggctggagcc cgtgcagttc    300 

ctcaggagtg tcttcgccaa cagcctggtg tacggcgcct ctgacagcaa cgtctatgac    360 

ctcctaaagg acctagagga aggcatccaa acgctgatgg ggaggctgga agatggcagc    420 

ccccggactg ggcagatctt caagcagacc tacagcaagt tcgacacaaa ctcacacaac    480 

gatgacgcac tactcaagaa ctacgggctg ctctactgct tcaggaagga catggacaag    540 

gtcgagacat tcctgcgcat cgtgcagtgc cgctctgtgg agggatcatg tggcttcttc    600 

ccaaccattc ccttatccag gctttttgac aacgctatgc tccgcgccca tcgtctgcac    660 

cagctggcct ttgacaccta ccaggagttt gaagaagcct atatcccaaa ggaacagaag    720 

tattcattcc tgcagaaccc ccagacctcc ctctgtttct cagagtctat tccgacaccc    780 

tccaacaggg aggaaacaca acagaaatcc aacctagagc tgctccgcat ctccctgctg    840 

ctcatccagt cgtggctgga gcccgtgcag ttcctcagga gtgtcttcgc caacagcctg    900 

gtgtacggcg cctctgacag caacgtctat gacctcctaa aggacctaga ggaaggcatc    960 

caaacgctga tggggaggct ggaagatggc agcccccgga ctgggcagat cttcaagcag   1020 

acctacagca agttcgacac aaactcacac aacgatgacg cactactcaa gaactacggg   1080 

ctgctctact gcttcaggaa ggacatggac aaggtcgaga cattcctgcg catcgtgcag   1140 

tgccgctctg tggagggatc c                                             1161 

 
           
             27  
             382  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            27 

Ser Cys Gly Phe Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn 
1               5                   10                  15 

Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr 
            20                  25                  30 

Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe 
        35                  40                  45 

Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr 
    50                  55                  60 

Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu 
65                  70                  75                  80 

Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe 
                85                  90                  95 

Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser 
            100                 105                 110 

Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu 
        115                 120                 125 

Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys 
    130                 135                 140 

Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu 
145                 150                 155                 160 

Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys 
                165                 170                 175 

Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser 
            180                 185                 190 

Cys Gly Phe Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala 
        195                 200                 205 

Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln 
    210                 215                 220 

Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu 
225                 230                 235                 240 

Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro 
                245                 250                 255 

Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg 
            260                 265                 270 

Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu 
        275                 280                 285 

Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn 
    290                 295                 300 

Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met 
305                 310                 315                 320 

Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln 
                325                 330                 335 

Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu 
            340                 345                 350 

Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val 
        355                 360                 365 

Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly 
    370                 375                 380 

 
           
             28  
             1152  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            28 

tgatcatgtg gcttcttccc aaccattccc ttatccaggc tttttgacaa cgctatgctc     60 

cgcgcccatc gtctgcacca gctggccttt gacacctacc aggagtttga agaagcctat    120 

atcccaaagg aacagaagta ttcattcctg cagaaccccc agacctccct ctgtttctca    180 

gagtctattc cgacaccctc caacagggag gaaacacaac agaaatccaa cctagagctg    240 

ctccgcatct ccctgctgct catccagtcg tggctggagc ccgtgcagtt cctcaggagt    300 

gtcttcgcca acagcctggt gtacggcgcc tctgacagca acgtctatga cctcctaaag    360 

gacctagagg aaggcatcca aacgctgatg gggaggctgg aagatggcag cccccggact    420 

gggcagatct tcaagcagac ctacagcaag ttcgacacaa actcacacaa cgatgacgca    480 

ctactcaaga actacgggct gctctactgc ttcaggaagg acatggacaa ggtcgagaca    540 

ttcctgcgca tcgtgcagtg ccgctctgtg gagggatcat gtggcttctt cccaaccatt    600 

cccttatcca ggctttttga caacgctatg ctccgcgccc atcgtctgca ccagctggcc    660 

tttgacacct accaggagtt tgaagaagcc tatatcccaa aggaacagaa gtattcattc    720 

ctgcagaacc cccagacctc cctctgtttc tcagagtcta ttccgacacc ctccaacagg    780 

gaggaaacac aacagaaatc caacctagag ctgctccgca tctccctgct gctcatccag    840 

tcgtggctgg agcccgtgca gttcctcagg agtgtcttcg ccaacagcct ggtgtacggc    900 

gcctctgaca gcaacgtcta tgacctccta aaggacctag aggaaggcat ccaaacgctg    960 

atggggaggc tggaagatgg cagcccccgg actgggcaga tcttcaagca gacctacagc   1020 

aagttcgaca caaactcaca caacgatgac gcactactca agaactacgg gctgctctac   1080 

tgcttcagga aggacatgga caaggtcgag acattcctgc gcatcgtgca gtgccgctct   1140 

gtggagggat cc                                                       1152 

 
           
             29  
             382  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            29 

Ser Cys Gly Phe Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn 
1               5                   10                  15 

Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr 
            20                  25                  30 

Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe 
        35                  40                  45 

Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr 
    50                  55                  60 

Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu 
65                  70                  75                  80 

Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe 
                85                  90                  95 

Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser 
            100                 105                 110 

Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu 
        115                 120                 125 

Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys 
    130                 135                 140 

Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu 
145                 150                 155                 160 

Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys 
                165                 170                 175 

Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser 
            180                 185                 190 

Cys Gly Phe Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala 
        195                 200                 205 

Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln 
    210                 215                 220 

Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu 
225                 230                 235                 240 

Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro 
                245                 250                 255 

Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg 
            260                 265                 270 

Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu 
        275                 280                 285 

Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn 
    290                 295                 300 

Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met 
305                 310                 315                 320 

Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln 
                325                 330                 335 

Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu 
            340                 345                 350 

Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val 
        355                 360                 365 

Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly 
    370                 375                 380 

 
           
             30  
             606  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            30 

catatgttcc caaccattcc cttatccagg ctttttgaca acgctatgct ccgcgcccat     60 

cgtctgcacc agctggcctt tgacacctac caggagtttg aagaagccta tatcccaaag    120 

gaacagaagt attcattcct gcagaacccc cagacctccc tctgtttctc agagtctatt    180 

ccgacaccct ccaacaggga ggaaacacaa cagaaatcca acctagagct gctccgcatc    240 

tccctgctgc tcatccagtc gtggctggag cccgtgcagt tcctcaggag tgtcttcgcc    300 

aacagcctgg tgtacggcgc ctctgacagc aacgtctatg acctcctaaa ggacctagag    360 

gaaggcatcc aaacgctgat ggggaggctg gaagatggca gcccccggac tgggcagatc    420 

ttcaagcaga cctacagcaa gttcgacaca aactcacaca acgatgacgc actactcaag    480 

aactacgggc tgctctactg cttcaggaag gacatggaca aggtcgagac attcctgcgc    540 

atcgtgcagt gccgctctgt ggagggatcc gaattccatt gatcatgtgg cttctagtag    600 

gtcgac                                                               606 

 
           
             31  
             1737  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            31 

catatgttcc caaccattcc cttatccagg ctttttgaca acgctatgct ccgcgcccat     60 

cgtctgcacc agctggcctt tgacacctac caggagtttg aagaagccta tatcccaaag    120 

gaacagaagt attcattcct gcagaacccc cagacctccc tctgtttctc agagtctatt    180 

ccgacaccct ccaacaggga ggaaacacaa cagaaatcca acctagagct gctccgcatc    240 

tccctgctgc tcatccagtc gtggctggag cccgtgcagt tcctcaggag tgtcttcgcc    300 

aacagcctgg tgtacggcgc ctctgacagc aacgtctatg acctcctaaa ggacctagag    360 

gaaggcatcc aaacgctgat ggggaggctg gaagatggca gcccccggac tgggcagatc    420 

ttcaagcaga cctacagcaa gttcgacaca aactcacaca acgatgacgc actactcaag    480 

aactacgggc tgctctactg cttcaggaag gacatggaca aggtcgagac attcctgcgc    540 

atcgtgcagt gccgctctgt ggagggatca tgtggcttct tcccaaccat tcccttatcc    600 

aggctttttg acaacgctat gctccgcgcc catcgtctgc accagctggc ctttgacacc    660 

taccaggagt ttgaagaagc ctatatccca aaggaacaga agtattcatt cctgcagaac    720 

ccccagacct ccctctgttt ctcagagtct attccgacac cctccaacag ggaggaaaca    780 

caacagaaat ccaacctaga gctgctccgc atctccctgc tgctcatcca gtcgtggctg    840 

gagcccgtgc agttcctcag gagtgtcttc gccaacagcc tggtgtacgg cgcctctgac    900 

agcaacgtct atgacctcct aaaggaccta gaggaaggca tccaaacgct gatggggagg    960 

ctggaagatg gcagcccccg gactgggcag atcttcaagc agacctacag caagttcgac   1020 

acaaactcac acaacgatga cgcactactc aagaactacg ggctgctcta ctgcttcagg   1080 

aaggacatgg acaaggtcga gacattcctg cgcatcgtgc agtgccgctc tgtggaggga   1140 

tcatgtggct tcttcccaac cattccctta tccaggcttt ttgacaacgc tatgctccgc   1200 

gcccatcgtc tgcaccagct ggcctttgac acctaccagg agtttgaaga agcctatatc   1260 

ccaaaggaac agaagtattc attcctgcag aacccccaga cctccctctg tttctcagag   1320 

tctattccga caccctccaa cagggaggaa acacaacaga aatccaacct agagctgctc   1380 

cgcatctccc tgctgctcat ccagtcgtgg ctggagcccg tgcagttcct caggagtgtc   1440 

ttcgccaaca gcctggtgta cggcgcctct gacagcaacg tctatgacct cctaaaggac   1500 

ctagaggaag gcatccaaac gctgatgggg aggctggaag atggcagccc ccggactggg   1560 

cagatcttca agcagaccta cagcaagttc gacacaaact cacacaacga tgacgcacta   1620 

ctcaagaact acgggctgct ctactgcttc aggaaggaca tggacaaggt cgagacattc   1680 

ctgcgcatcg tgcagtgccg ctctgtggag ggatcatgtg gcttctagta ggtcgac      1737 

 
           
             32  
             574  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            32 

Met Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg 
        195                 200                 205 

Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu 
    210                 215                 220 

Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro 
225                 230                 235                 240 

Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg 
                245                 250                 255 

Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu 
            260                 265                 270 

Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val 
        275                 280                 285 

Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp 
    290                 295                 300 

Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu 
305                 310                 315                 320 

Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser 
                325                 330                 335 

Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr 
            340                 345                 350 

Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe 
        355                 360                 365 

Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe Phe 
    370                 375                 380 

Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala 
385                 390                 395                 400 

His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu 
                405                 410                 415 

Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln 
            420                 425                 430 

Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu 
        435                 440                 445 

Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu 
    450                 455                 460 

Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe 
465                 470                 475                 480 

Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu 
                485                 490                 495 

Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu 
            500                 505                 510 

Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys 
        515                 520                 525 

Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly 
    530                 535                 540 

Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu 
545                 550                 555                 560 

Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
                565                 570 

 
           
             33  
             55  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            33 

taccatatga catgatcatg tggcttcggt ttcccaacca ttcccttatc caggc          55 

 
           
             34  
             591  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            34 

catatgacat gatcatgtgg cttcggtttc ccaaccattc ccttatccag gctttttgac     60 

aacgctatgc tccgcgccca tcgtctgcac cagctggcct ttgacaccta ccaggagttt    120 

gaagaagcct atatcccaaa ggaacagaag tattcattcc tgcagaaccc ccagacctcc    180 

ctctgtttct cagagtctat tccgacaccc tccaacaggg aggaaacaca acagaaatcc    240 

aacctagagc tgctccgcat ctccctgctg ctcatccagt cgtggctgga gcccgtgcag    300 

ttcctcagga gtgtcttcgc caacagcctg gtgtacggcg cctctgacag caacgtctat    360 

gacctcctaa aggacctaga ggaaggcatc caaacgctga tggggaggct ggaagatggc    420 

agcccccgga ctgggcagat cttcaagcag acctacagca agttcgacac aaactcacac    480 

aacgatgacg cactactcaa gaactacggg ctgctctact gcttcaggaa ggacatggac    540 

aaggtcgaga cattcctgcg catcgtgcag tgccgctctg tggagggatc c             591 

 
           
             35  
             192  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            35 

Ser Cys Gly Phe Gly Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp 
1               5                   10                  15 

Asn Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr 
            20                  25                  30 

Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser 
        35                  40                  45 

Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro 
    50                  55                  60 

Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu 
65                  70                  75                  80 

Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln 
                85                  90                  95 

Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp 
            100                 105                 110 

Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr 
        115                 120                 125 

Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe 
    130                 135                 140 

Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala 
145                 150                 155                 160 

Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp 
                165                 170                 175 

Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly 
            180                 185                 190 

 
           
             36  
             1158  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            36 

tgatcatgtg gcttcggttt cccaaccatt cccttatcca ggctttttga caacgctatg     60 

ctccgcgccc atcgtctgca ccagctggcc tttgacacct accaggagtt tgaagaagcc    120 

tatatcccaa aggaacagaa gtattcattc ctgcagaacc cccagacctc cctctgtttc    180 

tcagagtcta ttccgacacc ctccaacagg gaggaaacac aacagaaatc caacctagag    240 

ctgctccgca tctccctgct gctcatccag tcgtggctgg agcccgtgca gttcctcagg    300 

agtgtcttcg ccaacagcct ggtgtacggc gcctctgaca gcaacgtcta tgacctccta    360 

aaggacctag aggaaggcat ccaaacgctg atggggaggc tggaagatgg cagcccccgg    420 

actgggcaga tcttcaagca gacctacagc aagttcgaca caaactcaca caacgatgac    480 

gcactactca agaactacgg gctgctctac tgcttcagga aggacatgga caaggtcgag    540 

acattcctgc gcatcgtgca gtgccgctct gtggagggat catgtggctt cggtttccca    600 

accattccct tatccaggct ttttgacaac gctatgctcc gcgcccatcg tctgcaccag    660 

ctggcctttg acacctacca ggagtttgaa gaagcctata tcccaaagga acagaagtat    720 

tcattcctgc agaaccccca gacctccctc tgtttctcag agtctattcc gacaccctcc    780 

aacagggagg aaacacaaca gaaatccaac ctagagctgc tccgcatctc cctgctgctc    840 

atccagtcgt ggctggagcc cgtgcagttc ctcaggagtg tcttcgccaa cagcctggtg    900 

tacggcgcct ctgacagcaa cgtctatgac ctcctaaagg acctagagga aggcatccaa    960 

acgctgatgg ggaggctgga agatggcagc ccccggactg ggcagatctt caagcagacc   1020 

tacagcaagt tcgacacaaa ctcacacaac gatgacgcac tactcaagaa ctacgggctg   1080 

ctctactgct tcaggaagga catggacaag gtcgagacat tcctgcgcat cgtgcagtgc   1140 

cgctctgtgg agggatcc                                                 1158 

 
           
             37  
             384  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            37 

Ser Cys Gly Phe Gly Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp 
1               5                   10                  15 

Asn Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr 
            20                  25                  30 

Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser 
        35                  40                  45 

Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro 
    50                  55                  60 

Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu 
65                  70                  75                  80 

Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln 
                85                  90                  95 

Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp 
            100                 105                 110 

Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr 
        115                 120                 125 

Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe 
    130                 135                 140 

Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala 
145                 150                 155                 160 

Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp 
                165                 170                 175 

Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly 
            180                 185                 190 

Ser Cys Gly Phe Gly Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp 
        195                 200                 205 

Asn Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr 
    210                 215                 220 

Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser 
225                 230                 235                 240 

Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro 
                245                 250                 255 

Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu 
            260                 265                 270 

Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln 
        275                 280                 285 

Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp 
    290                 295                 300 

Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr 
305                 310                 315                 320 

Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe 
                325                 330                 335 

Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala 
            340                 345                 350 

Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp 
        355                 360                 365 

Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly 
    370                 375                 380 

 
           
             38  
             1743  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            38 

catatgttcc caaccattcc cttatccagg ctttttgaca acgctatgct ccgcgcccat     60 

cgtctgcacc agctggcctt tgacacctac caggagtttg aagaagccta tatcccaaag    120 

gaacagaagt attcattcct gcagaacccc cagacctccc tctgtttctc agagtctatt    180 

ccgacaccct ccaacaggga ggaaacacaa cagaaatcca acctagagct gctccgcatc    240 

tccctgctgc tcatccagtc gtggctggag cccgtgcagt tcctcaggag tgtcttcgcc    300 

aacagcctgg tgtacggcgc ctctgacagc aacgtctatg acctcctaaa ggacctagag    360 

gaaggcatcc aaacgctgat ggggaggctg gaagatggca gcccccggac tgggcagatc    420 

ttcaagcaga cctacagcaa gttcgacaca aactcacaca acgatgacgc actactcaag    480 

aactacgggc tgctctactg cttcaggaag gacatggaca aggtcgagac attcctgcgc    540 

atcgtgcagt gccgctctgt ggagggatca tgtggcttcg gtttcccaac cattccctta    600 

tccaggcttt ttgacaacgc tatgctccgc gcccatcgtc tgcaccagct ggcctttgac    660 

acctaccagg agtttgaaga agcctatatc ccaaaggaac agaagtattc attcctgcag    720 

aacccccaga cctccctctg tttctcagag tctattccga caccctccaa cagggaggaa    780 

acacaacaga aatccaacct agagctgctc cgcatctccc tgctgctcat ccagtcgtgg    840 

ctggagcccg tgcagttcct caggagtgtc ttcgccaaca gcctggtgta cggcgcctct    900 

gacagcaacg tctatgacct cctaaaggac ctagaggaag gcatccaaac gctgatgggg    960 

aggctggaag atggcagccc ccggactggg cagatcttca agcagaccta cagcaagttc   1020 

gacacaaact cacacaacga tgacgcacta ctcaagaact acgggctgct ctactgcttc   1080 

aggaaggaca tggacaaggt cgagacattc ctgcgcatcg tgcagtgccg ctctgtggag   1140 

ggatcatgtg gcttcggttt cccaaccatt cccttatcca ggctttttga caacgctatg   1200 

ctccgcgccc atcgtctgca ccagctggcc tttgacacct accaggagtt tgaagaagcc   1260 

tatatcccaa aggaacagaa gtattcattc ctgcagaacc cccagacctc cctctgtttc   1320 

tcagagtcta ttccgacacc ctccaacagg gaggaaacac aacagaaatc caacctagag   1380 

ctgctccgca tctccctgct gctcatccag tcgtggctgg agcccgtgca gttcctcagg   1440 

agtgtcttcg ccaacagcct ggtgtacggc gcctctgaca gcaacgtcta tgacctccta   1500 

aaggacctag aggaaggcat ccaaacgctg atggggaggc tggaagatgg cagcccccgg   1560 

actgggcaga tcttcaagca gacctacagc aagttcgaca caaactcaca caacgatgac   1620 

gcactactca agaactacgg gctgctctac tgcttcagga aggacatgga caaggtcgag   1680 

acattcctgc gcatcgtgca gtgccgctct gtggagggat catgtggctt ctagtaggtc   1740 

gac                                                                 1743 

 
           
             39  
             576  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            39 

Met Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

Gly Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
        195                 200                 205 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
    210                 215                 220 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
225                 230                 235                 240 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
                245                 250                 255 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
            260                 265                 270 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
        275                 280                 285 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
    290                 295                 300 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
305                 310                 315                 320 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
                325                 330                 335 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
            340                 345                 350 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
        355                 360                 365 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
    370                 375                 380 

Gly Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
385                 390                 395                 400 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
                405                 410                 415 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
            420                 425                 430 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
        435                 440                 445 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
    450                 455                 460 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
465                 470                 475                 480 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
                485                 490                 495 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
            500                 505                 510 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
        515                 520                 525 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
    530                 535                 540 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
545                 550                 555                 560 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
                565                 570                 575 

 
           
             40  
             39  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            40 

cgcggatcct catgagaagc cacagctgcc ctccacaga                            39 

 
           
             41  
             591  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            41 

catatgttcc caaccattcc cttatccagg ctttttgaca acgctatgct ccgcgcccat     60 

cgtctgcacc agctggcctt tgacacctac caggagtttg aagaagccta tatcccaaag    120 

gaacagaagt attcattcct gcagaacccc cagacctccc tctgtttctc agagtctatt    180 

ccgacaccct ccaacaggga ggaaacacaa cagaaatcca acctagagct gctccgcatc    240 

tccctgctgc tcatccagtc gtggctggag cccgtgcagt tcctcaggag tgtcttcgcc    300 

aacagcctgg tgtacggcgc ctctgacagc aacgtctatg acctcctaaa ggacctagag    360 

gaaggcatcc aaacgctgat ggggaggctg gaagatggca gcccccggac tgggcagatc    420 

ttcaagcaga cctacagcaa gttcgacaca aactcacaca acgatgacgc actactcaag    480 

aactacgggc tgctctactg cttcaggaag gacatggaca aggtcgagac attcctgcgc    540 

atcgtgcagt gccgctctgt ggagggcagc tgtggcttct catgaggatc c             591 

 
           
             42  
             193  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            42 

Met Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

Ser 

 
           
             43  
             50  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            43 

catgccatgg ggtggtggag gaagtttccc aaccattccc ttatccaggc                50 

 
           
             44  
             606  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            44 

ccatggggtg gtggaggaag tttcccaacc attcccttat ccaggctttt tgacaacgct     60 

atgctccgcg cccatcgtct gcaccagctg gcctttgaca cctaccagga gtttgaagaa    120 

gcctatatcc caaaggaaca gaagtattca ttcctgcaga acccccagac ctccctctgt    180 

ttctcagagt ctattccgac accctccaac agggaggaaa cacaacagaa atccaaccta    240 

gagctgctcc gcatctccct gctgctcatc cagtcgtggc tggagcccgt gcagttcctc    300 

aggagtgtct tcgccaacag cctggtgtac ggcgcctctg acagcaacgt ctatgacctc    360 

ctaaaggacc tagaggaagg catccaaacg ctgatgggga ggctggaaga tggcagcccc    420 

cggactgggc agatcttcaa gcagacctac agcaagttcg acacaaactc acacaacgat    480 

gacgcactac tcaagaacta cgggctgctc tactgcttca ggaaggacat ggacaaggtc    540 

gagacattcc tgcgcatcgt gcagtgccgc tctgtggagg gcagctgtgg cttctcatga    600 

ggatcc                                                               606 

 
           
             45  
             198  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            45 

Trp Gly Gly Gly Gly Ser Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe 
1               5                   10                  15 

Asp Asn Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp 
            20                  25                  30 

Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr 
        35                  40                  45 

Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile 
    50                  55                  60 

Pro Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu 
65                  70                  75                  80 

Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val 
                85                  90                  95 

Gln Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser 
            100                 105                 110 

Asp Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln 
        115                 120                 125 

Thr Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile 
    130                 135                 140 

Phe Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp 
145                 150                 155                 160 

Ala Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met 
                165                 170                 175 

Asp Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu 
            180                 185                 190 

Gly Ser Cys Gly Phe Ser 
        195 

 
           
             46  
             603  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            46 

ccatggggtg gtggaggaag tttcccaacc attcccttat ccaggctttt tgacaacgct     60 

atgctccgcg cccatcgtct gcaccagctg gcctttgaca cctaccagga gtttgaagaa    120 

gcctatatcc caaaggaaca gaagtattca ttcctgcaga acccccagac ctccctctgt    180 

ttctcagagt ctattccgac accctccaac agggaggaaa cacaacagaa atccaaccta    240 

gagctgctcc gcatctccct gctgctcatc cagtcgtggc tggagcccgt gcagttcctc    300 

aggagtgtct tcgccaacag cctggtgtac ggcgcctctg acagcaacgt ctatgacctc    360 

ctaaaggacc tagaggaagg catccaaacg ctgatgggga ggctggaaga tggcagcccc    420 

cggactgggc agatcttcaa gcagacctac agcaagttcg acacaaactc acacaacgat    480 

gacgcactac tcaagaacta cgggctgctc tactgcttca ggaaggacat ggacaaggtc    540 

gagacattcc tgcgcatcgt gcagtgccgc tctgtggagg gcagctgtgg cttctaggga    600 

tcc                                                                  603 

 
           
             47  
             197  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            47 

Trp Gly Gly Gly Gly Ser Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe 
1               5                   10                  15 

Asp Asn Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp 
            20                  25                  30 

Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr 
        35                  40                  45 

Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile 
    50                  55                  60 

Pro Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu 
65                  70                  75                  80 

Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val 
                85                  90                  95 

Gln Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser 
            100                 105                 110 

Asp Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln 
        115                 120                 125 

Thr Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile 
    130                 135                 140 

Phe Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp 
145                 150                 155                 160 

Ala Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met 
                165                 170                 175 

Asp Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu 
            180                 185                 190 

Gly Ser Cys Gly Phe 
        195 

 
           
             48  
             1200  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            48 

ccatggggtg gtggaggaag tttcccaacc attcccttat ccaggctttt tgacaacgct     60 

atgctccgcg cccatcgtct gcaccagctg gcctttgaca cctaccagga gtttgaagaa    120 

gcctatatcc caaaggaaca gaagtattca ttcctgcaga acccccagac ctccctctgt    180 

ttctcagagt ctattccgac accctccaac agggaggaaa cacaacagaa atccaaccta    240 

gagctgctcc gcatctccct gctgctcatc cagtcgtggc tggagcccgt gcagttcctc    300 

aggagtgtct tcgccaacag cctggtgtac ggcgcctctg acagcaacgt ctatgacctc    360 

ctaaaggacc tagaggaagg catccaaacg ctgatgggga ggctggaaga tggcagcccc    420 

cggactgggc agatcttcaa gcagacctac agcaagttcg acacaaactc acacaacgat    480 

gacgcactac tcaagaacta cgggctgctc tactgcttca ggaaggacat ggacaaggtc    540 

gagacattcc tgcgcatcgt gcagtgccgc tctgtggagg gcagctgtgg cttctcatgg    600 

ggtggtggag gaagtttccc aaccattccc ttatccaggc tttttgacaa cgctatgctc    660 

cgcgcccatc gtctgcacca gctggccttt gacacctacc aggagtttga agaagcctat    720 

atcccaaagg aacagaagta ttcattcctg cagaaccccc agacctccct ctgtttctca    780 

gagtctattc cgacaccctc caacagggag gaaacacaac agaaatccaa cctagagctg    840 

ctccgcatct ccctgctgct catccagtcg tggctggagc ccgtgcagtt cctcaggagt    900 

gtcttcgcca acagcctggt gtacggcgcc tctgacagca acgtctatga cctcctaaag    960 

gacctagagg aaggcatcca aacgctgatg gggaggctgg aagatggcag cccccggact   1020 

gggcagatct tcaagcagac ctacagcaag ttcgacacaa actcacacaa cgatgacgca   1080 

ctactcaaga actacgggct gctctactgc ttcaggaagg acatggacaa ggtcgagaca   1140 

ttcctgcgca tcgtgcagtg ccgctctgtg gagggcagct gtggcttctc atgaggatcc   1200 

 
           
             49  
             396  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            49 

Trp Gly Gly Gly Gly Ser Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe 
1               5                   10                  15 

Asp Asn Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp 
            20                  25                  30 

Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr 
        35                  40                  45 

Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile 
    50                  55                  60 

Pro Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu 
65                  70                  75                  80 

Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val 
                85                  90                  95 

Gln Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser 
            100                 105                 110 

Asp Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln 
        115                 120                 125 

Thr Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile 
    130                 135                 140 

Phe Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp 
145                 150                 155                 160 

Ala Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met 
                165                 170                 175 

Asp Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu 
            180                 185                 190 

Gly Ser Cys Gly Phe Ser Trp Gly Gly Gly Gly Ser Phe Pro Thr Ile 
        195                 200                 205 

Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala His Arg Leu 
    210                 215                 220 

His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile 
225                 230                 235                 240 

Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu 
                245                 250                 255 

Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr Gln 
            260                 265                 270 

Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln 
        275                 280                 285 

Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe Ala Asn Ser 
    290                 295                 300 

Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys Asp 
305                 310                 315                 320 

Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu Asp Gly Ser 
                325                 330                 335 

Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp Thr 
            340                 345                 350 

Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu Tyr 
        355                 360                 365 

Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu Arg Ile Val 
    370                 375                 380 

Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe Ser 
385                 390                 395 

 
           
             50  
             1185  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            50 

catatgttcc caaccattcc cttatccagg ctttttgaca acgctatgct ccgcgcccat     60 

cgtctgcacc agctggcctt tgacacctac caggagtttg aagaagccta tatcccaaag    120 

gaacagaagt attcattcct gcagaacccc cagacctccc tctgtttctc agagtctatt    180 

ccgacaccct ccaacaggga ggaaacacaa cagaaatcca acctagagct gctccgcatc    240 

tccctgctgc tcatccagtc gtggctggag cccgtgcagt tcctcaggag tgtcttcgcc    300 

aacagcctgg tgtacggcgc ctctgacagc aacgtctatg acctcctaaa ggacctagag    360 

gaaggcatcc aaacgctgat ggggaggctg gaagatggca gcccccggac tgggcagatc    420 

ttcaagcaga cctacagcaa gttcgacaca aactcacaca acgatgacgc actactcaag    480 

aactacgggc tgctctactg cttcaggaag gacatggaca aggtcgagac attcctgcgc    540 

atcgtgcagt gccgctctgt ggagggcagc tgtggcttct catggggtgg tggaggaagt    600 

ttcccaacca ttcccttatc caggcttttt gacaacgcta tgctccgcgc ccatcgtctg    660 

caccagctgg cctttgacac ctaccaggag tttgaagaag cctatatccc aaaggaacag    720 

aagtattcat tcctgcagaa cccccagacc tccctctgtt tctcagagtc tattccgaca    780 

ccctccaaca gggaggaaac acaacagaaa tccaacctag agctgctccg catctccctg    840 

ctgctcatcc agtcgtggct ggagcccgtg cagttcctca ggagtgtctt cgccaacagc    900 

ctggtgtacg gcgcctctga cagcaacgtc tatgacctcc taaaggacct agaggaaggc    960 

atccaaacgc tgatggggag gctggaagat ggcagccccc ggactgggca gatcttcaag   1020 

cagacctaca gcaagttcga cacaaactca cacaacgatg acgcactact caagaactac   1080 

gggctgctct actgcttcag gaaggacatg gacaaggtcg agacattcct gcgcatcgtg   1140 

cagtgccgct ctgtggaggg cagctgtggc ttctcatgag gatcc                   1185 

 
           
             51  
             391  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            51 

Met Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

Ser Trp Gly Gly Gly Gly Ser Phe Pro Thr Ile Pro Leu Ser Arg Leu 
        195                 200                 205 

Phe Asp Asn Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe 
    210                 215                 220 

Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys 
225                 230                 235                 240 

Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser 
                245                 250                 255 

Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu 
            260                 265                 270 

Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro 
        275                 280                 285 

Val Gln Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala 
    290                 295                 300 

Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile 
305                 310                 315                 320 

Gln Thr Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln 
                325                 330                 335 

Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp 
            340                 345                 350 

Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp 
        355                 360                 365 

Met Asp Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val 
    370                 375                 380 

Glu Gly Ser Cys Gly Phe Ser 
385                 390 

 
           
             52  
             1779  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            52 

catatgttcc caaccattcc cttatccagg ctttttgaca acgctatgct ccgcgcccat     60 

cgtctgcacc agctggcctt tgacacctac caggagtttg aagaagccta tatcccaaag    120 

gaacagaagt attcattcct gcagaacccc cagacctccc tctgtttctc agagtctatt    180 

ccgacaccct ccaacaggga ggaaacacaa cagaaatcca acctagagct gctccgcatc    240 

tccctgctgc tcatccagtc gtggctggag cccgtgcagt tcctcaggag tgtcttcgcc    300 

aacagcctgg tgtacggcgc ctctgacagc aacgtctatg acctcctaaa ggacctagag    360 

gaaggcatcc aaacgctgat ggggaggctg gaagatggca gcccccggac tgggcagatc    420 

ttcaagcaga cctacagcaa gttcgacaca aactcacaca acgatgacgc actactcaag    480 

aactacgggc tgctctactg cttcaggaag gacatggaca aggtcgagac attcctgcgc    540 

atcgtgcagt gccgctctgt ggagggcagc tgtggcttct catggggtgg tggaggaagt    600 

ttcccaacca ttcccttatc caggcttttt gacaacgcta tgctccgcgc ccatcgtctg    660 

caccagctgg cctttgacac ctaccaggag tttgaagaag cctatatccc aaaggaacag    720 

aagtattcat tcctgcagaa cccccagacc tccctctgtt tctcagagtc tattccgaca    780 

ccctccaaca gggaggaaac acaacagaaa tccaacctag agctgctccg catctccctg    840 

ctgctcatcc agtcgtggct ggagcccgtg cagttcctca ggagtgtctt cgccaacagc    900 

ctggtgtacg gcgcctctga cagcaacgtc tatgacctcc taaaggacct agaggaaggc    960 

atccaaacgc tgatggggag gctggaagat ggcagccccc ggactgggca gatcttcaag   1020 

cagacctaca gcaagttcga cacaaactca cacaacgatg acgcactact caagaactac   1080 

gggctgctct actgcttcag gaaggacatg gacaaggtcg agacattcct gcgcatcgtg   1140 

cagtgccgct ctgtggaggg cagctgtggc ttctcatggg gtggtggagg aagtttccca   1200 

accattccct tatccaggct ttttgacaac gctatgctcc gcgcccatcg tctgcaccag   1260 

ctggcctttg acacctacca ggagtttgaa gaagcctata tcccaaagga acagaagtat   1320 

tcattcctgc agaaccccca gacctccctc tgtttctcag agtctattcc gacaccctcc   1380 

aacagggagg aaacacaaca gaaatccaac ctagagctgc tccgcatctc cctgctgctc   1440 

atccagtcgt ggctggagcc cgtgcagttc ctcaggagtg tcttcgccaa cagcctggtg   1500 

tacggcgcct ctgacagcaa cgtctatgac ctcctaaagg acctagagga aggcatccaa   1560 

acgctgatgg ggaggctgga agatggcagc ccccggactg ggcagatctt caagcagacc   1620 

tacagcaagt tcgacacaaa ctcacacaac gatgacgcac tactcaagaa ctacgggctg   1680 

ctctactgct tcaggaagga catggacaag gtcgagacat tcctgcgcat cgtgcagtgc   1740 

cgctctgtgg agggcagctg tggcttctca tgaggatcc                          1779 

 
           
             53  
             589  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            53 

Met Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

Ser Trp Gly Gly Gly Gly Ser Phe Pro Thr Ile Pro Leu Ser Arg Leu 
        195                 200                 205 

Phe Asp Asn Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe 
    210                 215                 220 

Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys 
225                 230                 235                 240 

Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser 
                245                 250                 255 

Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu 
            260                 265                 270 

Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro 
        275                 280                 285 

Val Gln Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala 
    290                 295                 300 

Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile 
305                 310                 315                 320 

Gln Thr Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln 
                325                 330                 335 

Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp 
            340                 345                 350 

Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp 
        355                 360                 365 

Met Asp Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val 
    370                 375                 380 

Glu Gly Ser Cys Gly Phe Ser Trp Gly Gly Gly Gly Ser Phe Pro Thr 
385                 390                 395                 400 

Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala His Arg 
                405                 410                 415 

Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr 
            420                 425                 430 

Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser 
        435                 440                 445 

Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr 
    450                 455                 460 

Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile 
465                 470                 475                 480 

Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe Ala Asn 
                485                 490                 495 

Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys 
            500                 505                 510 

Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu Asp Gly 
        515                 520                 525 

Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp 
    530                 535                 540 

Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu 
545                 550                 555                 560 

Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu Arg Ile 
                565                 570                 575 

Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe Ser 
            580                 585 

 
           
             54  
             2370  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            54 

catatgttcc caaccattcc cttatccagg ctttttgaca acgctatgct ccgcgcccat     60 

cgtctgcacc agctggcctt tgacacctac caggagtttg aagaagccta tatcccaaag    120 

gaacagaagt attcattcct gcagaacccc cagacctccc tctgtttctc agagtctatt    180 

ccgacaccct ccaacaggga ggaaacacaa cagaaatcca acctagagct gctccgcatc    240 

tccctgctgc tcatccagtc gtggctggag cccgtgcagt tcctcaggag tgtcttcgcc    300 

aacagcctgg tgtacggcgc ctctgacagc aacgtctatg acctcctaaa ggacctagag    360 

gaaggcatcc aaacgctgat ggggaggctg gaagatggca gcccccggac tgggcagatc    420 

ttcaagcaga cctacagcaa gttcgacaca aactcacaca acgatgacgc actactcaag    480 

aactacgggc tgctctactg cttcaggaag gacatggaca aggtcgagac attcctgcgc    540 

atcgtgcagt gccgctctgt ggagggcagc tgtggcttct catggggtgg tggaggaagt    600 

ttcccaacca ttcccttatc caggcttttt gacaacgcta tgctccgcgc ccatcgtctg    660 

caccagctgg cctttgacac ctaccaggag tttgaagaag cctatatccc aaaggaacag    720 

aagtattcat tcctgcagaa cccccagacc tccctctgtt tctcagagtc tattccgaca    780 

ccctccaaca gggaggaaac acaacagaaa tccaacctag agctgctccg catctccctg    840 

ctgctcatcc agtcgtggct ggagcccgtg cagttcctca ggagtgtctt cgccaacagc    900 

ctggtgtacg gcgcctctga cagcaacgtc tatgacctcc taaaggacct agaggaaggc    960 

atccaaacgc tgatggggag gctggaagat ggcagccccc ggactgggca gatcttcaag   1020 

cagacctaca gcaagttcga cacaaactca cacaacgatg acgcactact caagaactac   1080 

gggctgctct actgcttcag gaaggacatg gacaaggtcg agacattcct gcgcatcgtg   1140 

cagtgccgct ctgtggaggg cagctgtggc ttctcatggg gtggtggagg aagtttccca   1200 

accattccct tatccaggct ttttgacaac gctatgctcc gcgcccatcg tctgcaccag   1260 

ctggcctttg acacctacca ggagtttgaa gaagcctata tcccaaagga acagaagtat   1320 

tcattcctgc agaaccccca gacctccctc tgtttctcag agtctattcc gacaccctcc   1380 

aacagggagg aaacacaaca gaaatccaac ctagagctgc tccgcatctc cctgctgctc   1440 

atccagtcgt ggctggagcc cgtgcagttc ctcaggagtg tcttcgccaa cagcctggtg   1500 

tacggcgcct ctgacagcaa cgtctatgac ctcctaaagg acctagagga aggcatccaa   1560 

acgctgatgg ggaggctgga agatggcagc ccccggactg ggcagatctt caagcagacc   1620 

tacagcaagt tcgacacaaa ctcacacaac gatgacgcac tactcaagaa ctacgggctg   1680 

ctctactgct tcaggaagga catggacaag gtcgagacat tcctgcgcat cgtgcagtgc   1740 

cgctctgtgg agggcagctg tggcttctca tggggtggtg gaggaagttt cccaaccatt   1800 

cccttatcca ggctttttga caacgctatg ctccgcgccc atcgtctgca ccagctggcc   1860 

tttgacacct accaggagtt tgaagaagcc tatatcccaa aggaacagaa gtattcattc   1920 

ctgcagaacc cccagacctc cctctgtttc tcagagtcta ttccgacacc ctccaacagg   1980 

gaggaaacac aacagaaatc caacctagag ctgctccgca tctccctgct gctcatccag   2040 

tcgtggctgg agcccgtgca gttcctcagg agtgtcttcg ccaacagcct ggtgtacggc   2100 

gcctctgaca gcaacgtcta tgacctccta aaggacctag aggaaggcat ccaaacgctg   2160 

atggggaggc tggaagatgg cagcccccgg actgggcaga tcttcaagca gacctacagc   2220 

aagttcgaca caaactcaca caacgatgac gcactactca agaactacgg gctgctctac   2280 

tgcttcagga aggacatgga caaggtcgag acattcctgc gcatcgtgca gtgccgctct   2340 

gtggagggca gctgtggctt ctagggatcc                                    2370 

 
           
             55  
             786  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            55 

Met Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

Ser Trp Gly Gly Gly Gly Ser Phe Pro Thr Ile Pro Leu Ser Arg Leu 
        195                 200                 205 

Phe Asp Asn Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe 
    210                 215                 220 

Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys 
225                 230                 235                 240 

Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser 
                245                 250                 255 

Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu 
            260                 265                 270 

Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro 
        275                 280                 285 

Val Gln Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala 
    290                 295                 300 

Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile 
305                 310                 315                 320 

Gln Thr Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln 
                325                 330                 335 

Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp 
            340                 345                 350 

Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp 
        355                 360                 365 

Met Asp Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val 
    370                 375                 380 

Glu Gly Ser Cys Gly Phe Ser Trp Gly Gly Gly Gly Ser Phe Pro Thr 
385                 390                 395                 400 

Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala His Arg 
                405                 410                 415 

Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr 
            420                 425                 430 

Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser 
        435                 440                 445 

Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr 
    450                 455                 460 

Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile 
465                 470                 475                 480 

Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe Ala Asn 
                485                 490                 495 

Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys 
            500                 505                 510 

Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu Asp Gly 
        515                 520                 525 

Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp 
    530                 535                 540 

Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu 
545                 550                 555                 560 

Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu Arg Ile 
                565                 570                 575 

Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe Ser Trp Gly Gly 
            580                 585                 590 

Gly Gly Ser Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala 
        595                 600                 605 

Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln 
    610                 615                 620 

Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu 
625                 630                 635                 640 

Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro 
                645                 650                 655 

Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg 
            660                 665                 670 

Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu 
        675                 680                 685 

Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn 
    690                 695                 700 

Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met 
705                 710                 715                 720 

Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln 
                725                 730                 735 

Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu 
            740                 745                 750 

Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val 
        755                 760                 765 

Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys 
    770                 775                 780 

Gly Phe 
785 

 
           
             56  
             33  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            56 

ttaccatgga ttgccggcgg cggcggatcc aat                                  33 

 
           
             57  
             36  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            57 

ttaccatgga tttgatcagg cggcggcgga tccaat                               36 

 
           
             58  
             36  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            58 

tgatcaggcg gcggcggatc aggcggcggc ggatcc                               36 

 
           
             59  
             10  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            59 

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 
1               5                   10 

 
           
             60  
             48  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            60 

gccggcggcg gcggatcagg cggcggcgga tcaggcggcg gcggatcc                  48 

 
           
             61  
             14  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            61 

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 
1               5                   10 

 
           
             62  
             43  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            62 

ggacatatgc tgtgatcatt cccaaccatt cccttatcca ggc                       43 

 
           
             63  
             41  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            63 

cgcgaattcg atccatggaa gccacagctg ccctccacag a                         41 

 
           
             64  
             36  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            64 

cgcgtcgacc tagaagccac agctgccctc cacaga                               36 

 
           
             65  
             602  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            65 

catatgctgt gatcattccc aaccattccc ttatccaggc tttttgacaa cgctatgctc     60 

cgcgcccatc gtctgcacca gctggccttt gacacctacc aggagtttga agaagcctat    120 

atcccaaagg aacagaagta ttcattcctg cagaaccccc agacctccct ctgtttctca    180 

gagtctattc cgacaccctc caacagggag gaaacacaac agaaatccaa cctagagctg    240 

ctccgcatct ccctgctgct catccagtcg tggctggagc ccgtgcagtt cctcaggagt    300 

gtcttcgcca acagcctggt gtacggcgcc tctgacagca acgtctatga cctcctaaag    360 

gacctagagg aaggcatcca aacgctgatg gggaggctgg aagatggcag cccccggact    420 

gggcagatct tcaagcagac ctacagcaag ttcgacacaa actcacacaa cgatgacgca    480 

ctactcaaga actacgggct gctctactgc ttcaggaagg acatggacaa ggtcgagaca    540 

ttcctgcgca tcgtgcagtg ccgctctgtg gagggcagct gtggcttcca tggatcgaat    600 

tc                                                                   602 

 
           
             66  
             192  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            66 

Ser Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

 
           
             67  
             600  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            67 

catatgctgt gatcattccc aaccattccc ttatccaggc tttttgacaa cgctatgctc     60 

cgcgcccatc gtctgcacca gctggccttt gacacctacc aggagtttga agaagcctat    120 

atcccaaagg aacagaagta ttcattcctg cagaaccccc agacctccct ctgtttctca    180 

gagtctattc cgacaccctc caacagggag gaaacacaac agaaatccaa cctagagctg    240 

ctccgcatct ccctgctgct catccagtcg tggctggagc ccgtgcagtt cctcaggagt    300 

gtcttcgcca acagcctggt gtacggcgcc tctgacagca acgtctatga cctcctaaag    360 

gacctagagg aaggcatcca aacgctgatg gggaggctgg aagatggcag cccccggact    420 

gggcagatct tcaagcagac ctacagcaag ttcgacacaa actcacacaa cgatgacgca    480 

ctactcaaga actacgggct gctctactgc ttcaggaagg acatggacaa ggtcgagaca    540 

ttcctgcgca tcgtgcagtg ccgctctgtg gagggcagct gtggcttcta ggtcgacgcg    600 

 
           
             68  
             192  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            68 

Ser Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

 
           
             69  
             639  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            69 

catatgctgt gatcattccc aaccattccc ttatccaggc tttttgacaa cgctatgctc     60 

cgcgcccatc gtctgcacca gctggccttt gacacctacc aggagtttga agaagcctat    120 

atcccaaagg aacagaagta ttcattcctg cagaaccccc agacctccct ctgtttctca    180 

gagtctattc cgacaccctc caacagggag gaaacacaac agaaatccaa cctagagctg    240 

ctccgcatct ccctgctgct catccagtcg tggctggagc ccgtgcagtt cctcaggagt    300 

gtcttcgcca acagcctggt gtacggcgcc tctgacagca acgtctatga cctcctaaag    360 

gacctagagg aaggcatcca aacgctgatg gggaggctgg aagatggcag cccccggact    420 

gggcagatct tcaagcagac ctacagcaag ttcgacacaa actcacacaa cgatgacgca    480 

ctactcaaga actacgggct gctctactgc ttcaggaagg acatggacaa ggtcgagaca    540 

ttcctgcgca tcgtgcagtg ccgctctgtg gagggcagct gtggcttcgg cggcggcgga    600 

tcaggcggcg gcggatcagg cggcggcgga tccgaattc                           639 

 
           
             70  
             206  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            70 

Ser Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 
        195                 200                 205 

 
           
             71  
             630  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            71 

catatgttcc caaccattcc cttatccagg ctttttgaca acgctatgct ccgcgcccat     60 

cgtctgcacc agctggcctt tgacacctac caggagtttg aagaagccta tatcccaaag    120 

gaacagaagt attcattcct gcagaacccc cagacctccc tctgtttctc agagtctatt    180 

ccgacaccct ccaacaggga ggaaacacaa cagaaatcca acctagagct gctccgcatc    240 

tccctgctgc tcatccagtc gtggctggag cccgtgcagt tcctcaggag tgtcttcgcc    300 

aacagcctgg tgtacggcgc ctctgacagc aacgtctatg acctcctaaa ggacctagag    360 

gaaggcatcc aaacgctgat ggggaggctg gaagatggca gcccccggac tgggcagatc    420 

ttcaagcaga cctacagcaa gttcgacaca aactcacaca acgatgacgc actactcaag    480 

aactacgggc tgctctactg cttcaggaag gacatggaca aggtcgagac attcctgcgc    540 

atcgtgcagt gccgctctgt ggagggcagc tgtggcttcg gcggcggcgg atcaggcggc    600 

ggcggatcag gcggcggcgg atccgaattc                                     630 

 
           
             72  
             206  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            72 

Met Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 
        195                 200                 205 

 
           
             73  
             1248  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            73 

tgatcattcc caaccattcc cttatccagg ctttttgaca acgctatgct ccgcgcccat     60 

cgtctgcacc agctggcctt tgacacctac caggagtttg aagaagccta tatcccaaag    120 

gaacagaagt attcattcct gcagaacccc cagacctccc tctgtttctc agagtctatt    180 

ccgacaccct ccaacaggga ggaaacacaa cagaaatcca acctagagct gctccgcatc    240 

tccctgctgc tcatccagtc gtggctggag cccgtgcagt tcctcaggag tgtcttcgcc    300 

aacagcctgg tgtacggcgc ctctgacagc aacgtctatg acctcctaaa ggacctagag    360 

gaaggcatcc aaacgctgat ggggaggctg gaagatggca gcccccggac tgggcagatc    420 

ttcaagcaga cctacagcaa gttcgacaca aactcacaca acgatgacgc actactcaag    480 

aactacgggc tgctctactg cttcaggaag gacatggaca aggtcgagac attcctgcgc    540 

atcgtgcagt gccgctctgt ggagggcagc tgtggcttcg gcggcggcgg atcaggcggc    600 

ggcggatcag gcggcggcgg atcattccca accattccct tatccaggct ttttgacaac    660 

gctatgctcc gcgcccatcg tctgcaccag ctggcctttg acacctacca ggagtttgaa    720 

gaagcctata tcccaaagga acagaagtat tcattcctgc agaaccccca gacctccctc    780 

tgtttctcag agtctattcc gacaccctcc aacagggagg aaacacaaca gaaatccaac    840 

ctagagctgc tccgcatctc cctgctgctc atccagtcgt ggctggagcc cgtgcagttc    900 

ctcaggagtg tcttcgccaa cagcctggtg tacggcgcct ctgacagcaa cgtctatgac    960 

ctcctaaagg acctagagga aggcatccaa acgctgatgg ggaggctgga agatggcagc   1020 

ccccggactg ggcagatctt caagcagacc tacagcaagt tcgacacaaa ctcacacaac   1080 

gatgacgcac tactcaagaa ctacgggctg ctctactgct tcaggaagga catggacaag   1140 

gtcgagacat tcctgcgcat cgtgcagtgc cgctctgtgg agggcagctg tggcttcggc   1200 

ggcggcggat caggcggcgg cggatcaggc ggcggcggat ccgaattc                1248 

 
           
             74  
             412  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            74 

Ser Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Phe 
        195                 200                 205 

Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala 
    210                 215                 220 

His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu 
225                 230                 235                 240 

Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln 
                245                 250                 255 

Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu 
            260                 265                 270 

Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu 
        275                 280                 285 

Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe 
    290                 295                 300 

Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu 
305                 310                 315                 320 

Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu 
                325                 330                 335 

Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys 
            340                 345                 350 

Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly 
        355                 360                 365 

Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu 
    370                 375                 380 

Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe Gly Gly 
385                 390                 395                 400 

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 
                405                 410 

 
           
             75  
             2445  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            75 

catatgttcc caaccattcc cttatccagg ctttttgaca acgctatgct ccgcgcccat     60 

cgtctgcacc agctggcctt tgacacctac caggagtttg aagaagccta tatcccaaag    120 

gaacagaagt attcattcct gcagaacccc cagacctccc tctgtttctc agagtctatt    180 

ccgacaccct ccaacaggga ggaaacacaa cagaaatcca acctagagct gctccgcatc    240 

tccctgctgc tcatccagtc gtggctggag cccgtgcagt tcctcaggag tgtcttcgcc    300 

aacagcctgg tgtacggcgc ctctgacagc aacgtctatg acctcctaaa ggacctagag    360 

gaaggcatcc aaacgctgat ggggaggctg gaagatggca gcccccggac tgggcagatc    420 

ttcaagcaga cctacagcaa gttcgacaca aactcacaca acgatgacgc actactcaag    480 

aactacgggc tgctctactg cttcaggaag gacatggaca aggtcgagac attcctgcgc    540 

atcgtgcagt gccgctctgt ggagggcagc tgtggcttcg gcggcggcgg atcaggcggc    600 

ggcggatcag gcggcggcgg atcattccca accattccct tatccaggct ttttgacaac    660 

gctatgctcc gcgcccatcg tctgcaccag ctggcctttg acacctacca ggagtttgaa    720 

gaagcctata tcccaaagga acagaagtat tcattcctgc agaaccccca gacctccctc    780 

tgtttctcag agtctattcc gacaccctcc aacagggagg aaacacaaca gaaatccaac    840 

ctagagctgc tccgcatctc cctgctgctc atccagtcgt ggctggagcc cgtgcagttc    900 

ctcaggagtg tcttcgccaa cagcctggtg tacggcgcct ctgacagcaa cgtctatgac    960 

ctcctaaagg acctagagga aggcatccaa acgctgatgg ggaggctgga agatggcagc   1020 

ccccggactg ggcagatctt caagcagacc tacagcaagt tcgacacaaa ctcacacaac   1080 

gatgacgcac tactcaagaa ctacgggctg ctctactgct tcaggaagga catggacaag   1140 

gtcgagacat tcctgcgcat cgtgcagtgc cgctctgtgg agggcagctg tggcttcggc   1200 

ggcggcggat caggcggcgg cggatcaggc ggcggcggat cattcccaac cattccctta   1260 

tccaggcttt ttgacaacgc tatgctccgc gcccatcgtc tgcaccagct ggcctttgac   1320 

acctaccagg agtttgaaga agcctatatc ccaaaggaac agaagtattc attcctgcag   1380 

aacccccaga cctccctctg tttctcagag tctattccga caccctccaa cagggaggaa   1440 

acacaacaga aatccaacct agagctgctc cgcatctccc tgctgctcat ccagtcgtgg   1500 

ctggagcccg tgcagttcct caggagtgtc ttcgccaaca gcctggtgta cggcgcctct   1560 

gacagcaacg tctatgacct cctaaaggac ctagaggaag gcatccaaac gctgatgggg   1620 

aggctggaag atggcagccc ccggactggg cagatcttca agcagaccta cagcaagttc   1680 

gacacaaact cacacaacga tgacgcacta ctcaagaact acgggctgct ctactgcttc   1740 

aggaaggaca tggacaaggt cgagacattc ctgcgcatcg tgcagtgccg ctctgtggag   1800 

ggcagctgtg gcttcggcgg cggcggatca ggcggcggcg gatcaggcgg cggcggatca   1860 

ttcccaacca ttcccttatc caggcttttt gacaacgcta tgctccgcgc ccatcgtctg   1920 

caccagctgg cctttgacac ctaccaggag tttgaagaag cctatatccc aaaggaacag   1980 

aagtattcat tcctgcagaa cccccagacc tccctctgtt tctcagagtc tattccgaca   2040 

ccctccaaca gggaggaaac acaacagaaa tccaacctag agctgctccg catctccctg   2100 

ctgctcatcc agtcgtggct ggagcccgtg cagttcctca ggagtgtctt cgccaacagc   2160 

ctggtgtacg gcgcctctga cagcaacgtc tatgacctcc taaaggacct agaggaaggc   2220 

atccaaacgc tgatggggag gctggaagat ggcagccccc ggactgggca gatcttcaag   2280 

cagacctaca gcaagttcga cacaaactca cacaacgatg acgcactact caagaactac   2340 

gggctgctct actgcttcag gaaggacatg gacaaggtcg agacattcct gcgcatcgtg   2400 

cagtgccgct ctgtggaggg cagctgtggc ttctaggtcg acgcg                   2445 

 
           
             76  
             810  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            76 

Met Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Phe 
        195                 200                 205 

Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala 
    210                 215                 220 

His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu 
225                 230                 235                 240 

Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln 
                245                 250                 255 

Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu 
            260                 265                 270 

Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu 
        275                 280                 285 

Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe 
    290                 295                 300 

Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu 
305                 310                 315                 320 

Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu 
                325                 330                 335 

Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys 
            340                 345                 350 

Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly 
        355                 360                 365 

Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu 
    370                 375                 380 

Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe Gly Gly 
385                 390                 395                 400 

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Phe Pro Thr 
                405                 410                 415 

Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala His Arg 
            420                 425                 430 

Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr 
        435                 440                 445 

Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser 
    450                 455                 460 

Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr 
465                 470                 475                 480 

Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile 
                485                 490                 495 

Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe Ala Asn 
            500                 505                 510 

Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys 
        515                 520                 525 

Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu Asp Gly 
    530                 535                 540 

Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp 
545                 550                 555                 560 

Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu 
                565                 570                 575 

Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu Arg Ile 
            580                 585                 590 

Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe Gly Gly Gly Gly 
        595                 600                 605 

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Phe Pro Thr Ile Pro 
    610                 615                 620 

Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala His Arg Leu His 
625                 630                 635                 640 

Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro 
                645                 650                 655 

Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys 
            660                 665                 670 

Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln 
        675                 680                 685 

Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser 
    690                 695                 700 

Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe Ala Asn Ser Leu 
705                 710                 715                 720 

Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu 
                725                 730                 735 

Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu Asp Gly Ser Pro 
            740                 745                 750 

Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn 
        755                 760                 765 

Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys 
    770                 775                 780 

Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu Arg Ile Val Gln 
785                 790                 795                 800 

Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
                805                 810 

 
           
             77  
             593  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            77 

catatgttcc caaccattcc cttatccagg ctttttgaca acgctatgct ccgcgcccat     60 

cgtctgcacc agctggcctt tgacacctac caggagtttg aagaagccta tatcccaaag    120 

gaacagaagt attcattcct gcagaacccc cagacctccc tctgtttctc agagtctatt    180 

ccgacaccct ccaacaggga ggaaacacaa cagaaatcca acctagagct gctccgcatc    240 

tccctgctgc tcatccagtc gtggctggag cccgtgcagt tcctcaggag tgtcttcgcc    300 

aacagcctgg tgtacggcgc ctctgacagc aacgtctatg acctcctaaa ggacctagag    360 

gaaggcatcc aaacgctgat ggggaggctg gaagatggca gcccccggac tgggcagatc    420 

ttcaagcaga cctacagcaa gttcgacaca aactcacaca acgatgacgc actactcaag    480 

aactacgggc tgctctactg cttcaggaag gacatggaca aggtcgagac attcctgcgc    540 

atcgtgcagt gccgctctgt ggagggcagc tgtggcttcc atggatcgaa ttc           593 

 
           
             78  
             192  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            78 

Met Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

 
           
             79  
             592  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            79 

aagctttccc aaccattccc ttatccaggc tttttgacaa cgctatgctc cgcgcccatc     60 

gtctgcacca gctggccttt gacacctacc aggagtttga agaagcctat atcccaaagg    120 

aacagaagta ttcattcctg cagaaccccc agacctccct ctgtttctca gagtctattc    180 

cgacaccctc caacagggag gaaacacaac agaaatccaa cctagagctg ctccgcatct    240 

ccctgctgct catccagtcg tggctggagc ccgtgcagtt cctcaggagt gtcttcgcca    300 

acagcctggt gtacggcgcc tctgacagca acgtctatga cctcctaaag gacctagagg    360 

aaggcatcca aacgctgatg gggaggctgg aagatggcag cccccggact gggcagatct    420 

tcaagcagac ctacagcaag ttcgacacaa actcacacaa cgatgacgca ctactcaaga    480 

actacgggct gctctactgc ttcaggaagg acatggacaa ggtcgagaca ttcctgcgca    540 

tcgtgcagtg ccgctctgtg gagggcagct gtggcttcca tggatcgaat tc            592 

 
           
             80  
             191  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            80 

Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg 
1               5                   10                  15 

Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu 
            20                  25                  30 

Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro 
        35                  40                  45 

Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg 
    50                  55                  60 

Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu 
65                  70                  75                  80 

Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val 
                85                  90                  95 

Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp 
            100                 105                 110 

Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu 
        115                 120                 125 

Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser 
    130                 135                 140 

Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr 
145                 150                 155                 160 

Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe 
                165                 170                 175 

Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

 
           
             81  
             587  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            81 

aagctttccc aaccattccc ttatccaggc tttttgacaa cgctatgctc cgcgcccatc     60 

gtctgcacca gctggccttt gacacctacc aggagtttga agaagcctat atcccaaagg    120 

aacagaagta ttcattcctg cagaaccccc agacctccct ctgtttctca gagtctattc    180 

cgacaccctc caacagggag gaaacacaac agaaatccaa cctagagctg ctccgcatct    240 

ccctgctgct catccagtcg tggctggagc ccgtgcagtt cctcaggagt gtcttcgcca    300 

acagcctggt gtacggcgcc tctgacagca acgtctatga cctcctaaag gacctagagg    360 

aaggcatcca aacgctgatg gggaggctgg aagatggcag cccccggact gggcagatct    420 

tcaagcagac ctacagcaag ttcgacacaa actcacacaa cgatgacgca ctactcaaga    480 

actacgggct gctctactgc ttcaggaagg acatggacaa ggtcgagaca ttcctgcgca    540 

tcgtgcagtg ccgctctgtg gagggcagct gtggcttcta gggatcc                  587 

 
           
             82  
             191  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            82 

Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg 
1               5                   10                  15 

Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu 
            20                  25                  30 

Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro 
        35                  40                  45 

Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg 
    50                  55                  60 

Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu 
65                  70                  75                  80 

Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val 
                85                  90                  95 

Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp 
            100                 105                 110 

Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu 
        115                 120                 125 

Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser 
    130                 135                 140 

Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr 
145                 150                 155                 160 

Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe 
                165                 170                 175 

Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

 
           
             83  
             1165  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            83 

aagctttccc aaccattccc ttatccaggc tttttgacaa cgctatgctc cgcgcccatc     60 

gtctgcacca gctggccttt gacacctacc aggagtttga agaagcctat atcccaaagg    120 

aacagaagta ttcattcctg cagaaccccc agacctccct ctgtttctca gagtctattc    180 

cgacaccctc caacagggag gaaacacaac agaaatccaa cctagagctg ctccgcatct    240 

ccctgctgct catccagtcg tggctggagc ccgtgcagtt cctcaggagt gtcttcgcca    300 

acagcctggt gtacggcgcc tctgacagca acgtctatga cctcctaaag gacctagagg    360 

aaggcatcca aacgctgatg gggaggctgg aagatggcag cccccggact gggcagatct    420 

tcaagcagac ctacagcaag ttcgacacaa actcacacaa cgatgacgca ctactcaaga    480 

actacgggct gctctactgc ttcaggaagg acatggacaa ggtcgagaca ttcctgcgca    540 

tcgtgcagtg ccgctctgtg gagggcagct gtggcttctt cccaaccatt cccttatcca    600 

ggctttttga caacgctatg ctccgcgccc atcgtctgca ccagctggcc tttgacacct    660 

accaggagtt tgaagaagcc tatatcccaa aggaacagaa gtattcattc ctgcagaacc    720 

cccagacctc cctctgtttc tcagagtcta ttccgacacc ctccaacagg gaggaaacac    780 

aacagaaatc caacctagag ctgctccgca tctccctgct gctcatccag tcgtggctgg    840 

agcccgtgca gttcctcagg agtgtcttcg ccaacagcct ggtgtacggc gcctctgaca    900 

gcaacgtcta tgacctccta aaggacctag aggaaggcat ccaaacgctg atggggaggc    960 

tggaagatgg cagcccccgg actgggcaga tcttcaagca gacctacagc aagttcgaca   1020 

caaactcaca caacgatgac gcactactca agaactacgg gctgctctac tgcttcagga   1080 

aggacatgga caaggtcgag acattcctgc gcatcgtgca gtgccgctct gtggagggca   1140 

gctgtggctt ccatggatcg aattc                                         1165 

 
           
             84  
             191  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            84 

Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg 
1               5                   10                  15 

Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu 
            20                  25                  30 

Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro 
        35                  40                  45 

Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg 
    50                  55                  60 

Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu 
65                  70                  75                  80 

Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val 
                85                  90                  95 

Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp 
            100                 105                 110 

Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu 
        115                 120                 125 

Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser 
    130                 135                 140 

Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr 
145                 150                 155                 160 

Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe 
                165                 170                 175 

Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190 

 
           
             85  
             2307  
             DNA  
             Artificial  
             
               synthetic sequence  
             
           
            85 

catatgttcc caaccattcc cttatccagg ctttttgaca acgctatgct ccgcgcccat     60 

cgtctgcacc agctggcctt tgacacctac caggagtttg aagaagccta tatcccaaag    120 

gaacagaagt attcattcct gcagaacccc cagacctccc tctgtttctc agagtctatt    180 

ccgacaccct ccaacaggga ggaaacacaa cagaaatcca acctagagct gctccgcatc    240 

tccctgctgc tcatccagtc gtggctggag cccgtgcagt tcctcaggag tgtcttcgcc    300 

aacagcctgg tgtacggcgc ctctgacagc aacgtctatg acctcctaaa ggacctagag    360 

gaaggcatcc aaacgctgat ggggaggctg gaagatggca gcccccggac tgggcagatc    420 

ttcaagcaga cctacagcaa gttcgacaca aactcacaca acgatgacgc actactcaag    480 

aactacgggc tgctctactg cttcaggaag gacatggaca aggtcgagac attcctgcgc    540 

atcgtgcagt gccgctctgt ggagggcagc tgtggcttct tcccaaccat tcccttatcc    600 

aggctttttg acaacgctat gctccgcgcc catcgtctgc accagctggc ctttgacacc    660 

taccaggagt ttgaagaagc ctatatccca aaggaacaga agtattcatt cctgcagaac    720 

ccccagacct ccctctgttt ctcagagtct attccgacac cctccaacag ggaggaaaca    780 

caacagaaat ccaacctaga gctgctccgc atctccctgc tgctcatcca gtcgtggctg    840 

gagcccgtgc agttcctcag gagtgtcttc gccaacagcc tggtgtacgg cgcctctgac    900 

agcaacgtct atgacctcct aaaggaccta gaggaaggca tccaaacgct gatggggagg    960 

ctggaagatg gcagcccccg gactgggcag atcttcaagc agacctacag caagttcgac   1020 

acaaactcac acaacgatga cgcactactc aagaactacg ggctgctcta ctgcttcagg   1080 

aaggacatgg acaaggtcga gacattcctg cgcatcgtgc agtgccgctc tgtggagggc   1140 

agctgtggct tcttcccaac cattccctta tccaggcttt ttgacaacgc tatgctccgc   1200 

gcccatcgtc tgcaccagct ggcctttgac acctaccagg agtttgaaga agcctatatc   1260 

ccaaaggaac agaagtattc attcctgcag aacccccaga cctccctctg tttctcagag   1320 

tctattccga caccctccaa cagggaggaa acacaacaga aatccaacct agagctgctc   1380 

cgcatctccc tgctgctcat ccagtcgtgg ctggagcccg tgcagttcct caggagtgtc   1440 

ttcgccaaca gcctggtgta cggcgcctct gacagcaacg tctatgacct cctaaaggac   1500 

ctagaggaag gcatccaaac gctgatgggg aggctggaag atggcagccc ccggactggg   1560 

cagatcttca agcagaccta cagcaagttc gacacaaact cacacaacga tgacgcacta   1620 

ctcaagaact acgggctgct ctactgcttc aggaaggaca tggacaaggt cgagacattc   1680 

ctgcgcatcg tgcagtgccg ctctgtggag ggcagctgtg gcttcttccc aaccattccc   1740 

ttatccaggc tttttgacaa cgctatgctc cgcgcccatc gtctgcacca gctggccttt   1800 

gacacctacc aggagtttga agaagcctat atcccaaagg aacagaagta ttcattcctg   1860 

cagaaccccc agacctccct ctgtttctca gagtctattc cgacaccctc caacagggag   1920 

gaaacacaac agaaatccaa cctagagctg ctccgcatct ccctgctgct catccagtcg   1980 

tggctggagc ccgtgcagtt cctcaggagt gtcttcgcca acagcctggt gtacggcgcc   2040 

tctgacagca acgtctatga cctcctaaag gacctagagg aaggcatcca aacgctgatg   2100 

gggaggctgg aagatggcag cccccggact gggcagatct tcaagcagac ctacagcaag   2160 

ttcgacacaa actcacacaa cgatgacgca ctactcaaga actacgggct gctctactgc   2220 

ttcaggaagg acatggacaa ggtcgagaca ttcctgcgca tcgtgcagtg ccgctctgtg   2280 

gagggcagct gtggcttcta gggatcc                                       2307 

 
           
             86  
             192  
             PRT  
             Artificial  
             
               synthetic sequence  
             
           
            86 

Met Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu 
1               5                   10                  15 

Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe 
            20                  25                  30 

Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn 
        35                  40                  45 

Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn 
    50                  55                  60 

Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser 
65                  70                  75                  80 

Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser 
                85                  90                  95 

Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr 
            100                 105                 110 

Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg 
        115                 120                 125 

Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr 
    130                 135                 140 

Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn 
145                 150                 155                 160 

Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr 
                165                 170                 175 

Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 
            180                 185                 190