PATENT ABSTRACT
This invention relates to the development of a mammalian expression vector, under which expression of the structural genes of western equine encephalitis virus have been placed under the control of an eucaryotic promoter. When the recombinant vector is administered to mammalian cell culture or using a cell-free transcription/translation system, in vitro, authentic structural proteins of western equine encephalitis virus are produced as verified by reactivity with monoclonal antibodies developed to western equine encephalitis virus. When the recombinant DNA molecule is administered in vivo, a protective immune response is induced, thereby enhancing protection of the individual against subsequent infection by western equine encephalitis virus. In a similar manner, DNA vaccines to related alphaviruses (Venezuelan and eastern equine encephalitis viruses) could also be developed.

PATENT DESCRIPTION
FIELD OF THE INVENTION  
         [0001]    This invention relates to the cloning, sequencing and expression of the structural genes of western equine encephalitis (WEE) virus strain 71V-1658 and the development and use of the DNA-based vaccine against WEE.  
         BACKGROUND OF THE INVENTION  
       LIST OF PRIOR ART LITERATURES  
         [0002]    Ausubel, F. M., et al, editors. (1995).  Current Protocols in Molecular Biology,  New York: John Wiley &amp; Sons.  
           [0003]    Bell, J. R., Bond, M. W., Nukapiller, M. B., Strauss, E. G., Strauss, J. H., Yamamoto, K, &amp; Simizu, B. (1983). Structural proteins of western equine encephalitis virus: amino acid compositions and N-terminal sequences.  Journal of Virology  45, 708-714.  
           [0004]    Bird, B. R. &amp; Forrester, F. T. (1981).  Basic Laboratory Techniques In Cell Culture.  Atlanta: U.S. Department of Health and Human Services, Centers for Disease Control.  
           [0005]    Calisher, C. H. &amp; Karabatsos, N. (1988). Arbovirus serogroups: definition and geographic distribution. In  The Arboviruses: Epidemiology and Ecology,  Vol. I,.pp. 19-57. Edited by T. P Monath. CRC Press: Boca Raton, Fla.  
           [0006]    Calisher, C. H., Shope, R. E, Brandt, W., Casals, J., Karabatsos, N., Murphy, F. A., Tesh, R. B., &amp; Wiebe, M. E. (1980). Proposed antigenic classification of registered arbovirusess.  Intervirology  14, 229-232.  
           [0007]    Calisher, C. H., Karabatsos, N., Lazuick, J. S. Monath, T. P., &amp; Wolff, K. L. (1988). Reevaluation of the western equine encephalitis antigenic complex of alphaviruses (family Togaviridae) as determined by neutralization tests.  American Journal of Tropical Medicine and Hygiene  38, 447-452.  
           [0008]    Cilnis, M. J., Kang, W. &amp; Weaver, S. C. (1996). Genetic conservation of Highlands J viruses.  Virology  218, 343-351.  
           [0009]    Frohman, M. A., Dush, M. K &amp; Martin, G. R. (1988). Rapid production of full-length cDNAs from rare transcripts: Amplification using a single gene-specific oligonucleotide primer.  Proceedings of the National Academy of Science USA  85, 8998-9002.  
           [0010]    Hahn, C. S., Lustig, S., Strauss, E. G. &amp; Strauss, J. H. (1988). Western Equine Encephalitis virus is a recombinant virus.  Proceedings of the National Academy of Science USA  85, 5997-6001.  
           [0011]    Johnson, R. E. &amp; Peters, C. J. (1996). Alphaviruses. In  Fields Virology,  3rd edn, pp. 843-898. Edited by B. N. Fields, et al., New York: Raven Press.  
           [0012]    Kuhn, R., Hong, Z. &amp; Strauss, J. H. (1990). Mutagenesis of the 3′ nontranslated region of Sindbis virus RNA.  Journal of Virology  64, 1465-1476.  
           [0013]    Kuhn, R. J., Niesters, H. G. M., Hong, Z. &amp; Strauss, J. H. (1991). Infectious RNA transcripts from Ross River virus cDNA clones and the construction and characterization of defined chimeras with Sindbis.  Virology  182, 430-441.  
           [0014]    Krieg, A. M., Yi, A.-K., Schorr, J. and Davis, H. L. (1998). The role of CpG dinucleotides in DNA vaccines.  Trends Microbiol.  6, 23-27.  
           [0015]    McCluskie, M. J., Davies, H. L. (1999). Novel strategies using DNA for the induction of mucosal immunity.  Critic. Rev. in Immunol.  19, 303-329.  
           [0016]    Ou, J.-H., Trent, D. W. &amp; Strauss, J. H. (1982). The 3′ non-coding regions of alphavirus RNAs contain repeating sequences. Journal of Molecular Biology 156, 719-730.  
           [0017]    Ou, J-H., Strauss, E. G. &amp; Strauss, J. H. (1983). The 5′ terminal sequences of the genomic RNAs of several alphaviruses. Journal of Molecular Biology 168, 1-15.  
           [0018]    Pardoll, D R, Beckering, A M. (1997). Exposing the immunology of naked DNA vaccines.  Immunity  3;165-169.  
           [0019]    Pfeffer, M., Proebster, B., Kinney, R. M. &amp; Kaaden, O-R. (1997). Genus-specific detection of alphaviruses by a semi-nested reverse transcription reaction. American Journal of tropical Medicine and Hygiene 57, 709-718.  
           [0020]    Pfeffer, M., Kinney, R. M. &amp; Kaaden, O-R. (1998). The alphavirus 3′-nontranslated region: Size heterogeneity and arrangement of repeated sequence elements. Virology 240, 100-108.  
           [0021]    Prayaga, S. K., Fuller, D. H., Haynes, J. R. &amp; Murphey-Corb, M. (1995). Particle-mediated nucleic acid immunization.  Vaccines  95, 105-109.  
           [0022]    Reisen, W. K &amp; Monath, T. P. (1988). Western equine encephalomyelitis, pp. 89-137 . In The Arboviruses: Epidemiology and Ecology, Vol. V.  Edited by T. P. Monath. CRC Press: Boca Raton, Fla.  
           [0023]    Robinson, H. L., Feltquate, D. M., Morin, M. J., Haynes, J. R., Webster, R. G. (1995). DNA vaccines: A new approach to immunization.  Vaccine  95:69-75.  
           [0024]    Sambrook, J., Fritsch, E. F., &amp; Maniatis, T. (1989).  Molecular Cloning, a Laboratory Manual,  2 nd edn.  Cold Spring Harbor: Cold Spring Harbor Laboratory.  
           [0025]    Schlesinger, S. &amp; Schlesinger, M. J. (1996). Togaviridae: The viruses and their replication, In  Fields Virology,  3rd edn, pp. 825-841. Edited by B. N. Fields, et al. New York: Raven Press.  
           [0026]    Strauss, J. H., &amp; Strauss, E. G. (1988). Evolution of RNA viruses.  Annual Review of Microbiology  42, 657-683.  
           [0027]    Strauss, J. H., &amp; Strauss, E. G. (1994). The alphaviruses: gene expression, replication, and evolution.  Microbiological Review  58, 491-562.  
           [0028]    Strauss, E. G., Rice, C. M. &amp; Strauss, J. H. (1983). Sequence coding for the alphavirus nonstructural proteins is interrupted by an opal termination codon.  Proceedings of the National Academy of Science USA  80, 5271-5275.  
           [0029]    Strauss, E. G., Rice, C. M. &amp; Strauss, J. H. (1984). Complete nucleotide sequence of the genomic RNA of Sindbis virus.  Virology  133, 92-110.  
           [0030]    Trent, D. W., &amp; Grant, J. A. (1980). A comparison of new world alphaviruses in the western equine encephalomyelitis complex by immunochemical and oligonucleotide fingerprint techniques.  Journal or General Virology  47:261-282.  
           [0031]    Weaver, S. C., Hagenbaugh, A., Bellew, L. A., Netesov, S. V., Volchokov, V. I., Chang, G.-J J., Clarke, D. K., Gousset, L., Scott, T. W., Trent, D. W. &amp; Holland, J. J. (1993). A comparison of the nucleotide seqeunces of eastern and western equine encephalomyelitis viruses with those of other alphaviruses and related RNA viruses.  Virology  197, 375-390.  
           [0032]    Weaver, S. C., Kang, W, Shirako, Y., Rumenapf, T., Strauss, E. G. &amp; Strauss, J. H. (1997) Recombinational history and molecular evolution of western equine encephalomyelitis complex alphaviruses.  Journal of Virology  71,613-623.  
           [0033]    Wolff, J. A., Malone, R. W., Williams, P., Chong, W., Aesasi, G., Jani, A., Felgner, P. L. (1990). Direct gene transfer into mouse muscle in vivo.  Science  247,1465-1468.  
           [0034]    The alphaviruses are a group of about 27 enveloped viruses with a positive sense, nonsegmented single-stranded RNA genome (Calisher et al., 1980; Strauss and Strauss, 1988). The alphavirus disclosed in this invention, western equine encephalitis virus (WEE), is a member of the WEE antigenic complex and is serologically related to the Sindbis (SIN), Highlands J (HJ), Fort Morgan, Buggy Creek, and Aura viruses (Calisher &amp; Karabatsos, 1988; Calisher et al., 1988). WEE is endemic in western North America and strains/varieties have been isolated from Argentina (AG80-646), Brazil (BeAr 102091) and the former Soviet Union (Y62-33) (Johnson and Peters, 1996; Weaver et al., 1997). In nature, WEE is transmitted from its amplifying hosts or reservoir in wild birds, to man and horses, by mosquitoes ( Culex tarsalis  being the principal vector). While the endemic cycle has resulted in only a limited number of human infections in recent years, in the past, major epidemics of WEE have been recorded. The most extensive epidemic, including 3,336 recognized human cases and 300,000 cases of encephalitis in horses and mules, occurred in the western United States and Canada in 1941 (Reisen &amp; Monath, 1988; Johnson and Peters, 1996).  
           [0035]    All alphaviruses share a number of structural, sequence, and functional similarities, including a genome with two polyprotein gene clusters (reviewed in Strauss &amp; Strauss, 1994; Schlesinger &amp; Schlesinger 1996). The genomic organization of these viruses is conserved (see FIG. 1), with the nonstructural proteins translated directly from the 5′ two-thirds of the genomic RNA. A subgenomic positive-stranded RNA (the 26S RNA), is identical to the 3′ one-third of the genomic RNA and serves as the translational template for the structural proteins (capsid, E3, E2,6K and E1).  
           [0036]    The nonstructural proteins (nsP1, nsP2, nsP3 and nsP4) are also synthesized as a polyprotein and processed into the four nsPs by a nsP2 protease. Two versions of the nonstructural polyprotein are synthesized in alphavirus-infected cells, due to frequent readthrough of an opal codon between the nsP3 and nsP4 genes in several alphaviruses (Strauss et al., 1983). The nsPs function in a complex with host factors to replicate the genome and transcribe the subgenomic mRNA. Alphaviruses have characteristic conserved sequences at the extreme 5′ and 3′ domains and the intergenic region (Ou et al., 1982, 1983; Pfeffer et al., 1998). These conserved domains are required for viral growth and replication and are believed to be important in promotion of protein synthesis and the initiation of RNA-dependent RNA polymerase activity.  
           [0037]    The relationship of different WEE isolates to each other has been demonstrated using neutralization tests (Calisher et al., 1988). Additionally, several strains of WEE were typed by oligonucleotide fingerprinting, and found to have greater than 90% nt homology (Trent &amp; Grant, 1980). The N-terminal sequences of the nucleocapsid, and the E1 and E2 glycoproteins have been determined by Edman degradation, and the E1 and E2 proteins were found to have 82% and 71% homology, respectively, to SIN (Bell et al, 1983). Hahn et al. (1988) sequenced the 26S region of WEE strain BFS1703. They proposed that WEE originated as a hybrid virus, formed by recombination of an EEE and a Sindbis-like virus, most likely during a co-infection event. They suggested that two crossover events occurred, one within the E3 gene, the other within the 3′ nontranslated terminal region (NTR), resulting in a virus whose nonstructural domain, intragenic region, and capsid protein are similar to EEE, with envelope proteins showing homology to SIN.  
           [0038]    Weaver et al. (1993) sequenced part of the nonstructural domain (nsP2 and nsP3 genes) of strain 5614, demonstrating this area also shows homology to EEE. Short regions within the nsP4 gene and the E1 protein/3′ NTR have been determined for many WEE strains, allowing a preliminary assessment of the nucleic acid phylogenetic relationships within the WEE antigenic complex (Weaver et al., 1997). Serological studies (Calisher et al., 1988) and preliminary sequence determination (Cilnis et al., 1996; Weaver et al., 1997) of the HJ genome suggests this is another closely related virus, and most likely a descendant of the same recombinant viral ancestor as modem WEE.  
           [0039]    A highly conserved region of the alphavirus nsP1 gene has been identified, and proved suitable for use in a polymerase chain reaction (PCR)-based genetic assay for alphaviruses, including WEE (Pfeffer et al., 1997). Phylogenetic analysis of this PCR fragment yielded similar results to those obtained by Weaver et al., (1997) for a PCR fragment in the nsP4 gene.  
           [0040]    In terms of therapy or prophylaxis, there are very limited possibilities. An inactivated vaccine to WEE is under investigational new drug (IND) status. The vaccine uses formalin-inactivation of cell culture supernatants from WEE-infected tissue culture. It requires a minimum of 3 doses, yearly monitoring of antibody titer and possible boosters. Its effectiveness in the protection against an aerosol challenge of WEE has yet to be established. A WEE live attenuated vaccine based on an infectious clone is under development (J. Smith, personnel communication). The area of DNA immunization is relatively new, and has been reviewed in Hassett and Whitton, 1996; Donnelly et al, 1997. Similar to live, attenuated vaccines, DNA vaccines are known to stimulate both humoral and cellular immune responses (Pardoll and Backering, 1997; McCuskie and Davies, 1999). Much of the focus has been on methods to deliver and efficiently express the cloned products. Intramuscular administration of DNA has been one of the original methods used (Wolff et al, 1990). A second method uses ballistic delivery of DNA coated gold particles, using high pressure helium gas to propel the particles into the epidermis and dermis of animals (Prayaga et al, 1995, reviewed by Robinson et al, 1995).  
           [0041]    The Applicant identified a number of related areas of research, including the development of subunit vaccines to WEE. In the present invention, the Applicant disclosed the cloning, sequencing and expression of the structural genes of a WEE virus (strain 71V-1658), as described in Netolitzky et al., (2000) “Complete genomic RNA sequence of western equine encephalitis virus and expression of the structural genes.”  Journal of General Virology  81, 151-159, which is herein incorporated by reference. The DNA construct (pCXH-3), and a second construct (pVHX-6) were used in DNA immunization studies in a mouse model for protection against intranasal administered WEE.  
         SUMMARY OF THE INVENTION  
         [0042]    The present invention is directed to the development of a DNA-subunit vaccine to the WEE virus and its use against such virus. More specifically, DNA to structural components of the WEE virus are expressed and used as the subunit vaccine.  
           [0043]    The present invention provides for the complete nucleotide sequence of WEE strain 71V-1658. Two novel cDNA clones, pCXH-3 and pVHX-6 are also disclosed as effective vectors for gene expression.  
           [0044]    The present invention also provides the complete nucleotide sequence for the structural gene pcDWXH-7.  
           [0045]    It further provides for a process for preparing a recombinant DNA vaccine against WEE virus, comprising cloning and sequencing of 26S region of a WEE virus strain 71V-1658 under conditions suitable to effect in vitro transcription and translation of the functional recombinant DNA expression vector pCXH-3 and pVHX-6. 
       
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0046]    [0046]FIG. 1. Diagram showing the WEE 71V-1658 sequencing strategy. The location of PCR probe sequences used to screen the WEE cDNA library are also indicated, along with the genomic organization of the virus.  
         [0047]    [0047]FIG. 2. Multiple sequence alignment.  
         [0048]    [0048]FIG. 3. Stem loop structures in the 5′ NTR.  
         [0049]    [0049]FIG. 4. Stem loop structures in the 3′ NTR.  
         [0050]    [0050]FIG. 5. Phylogenetic relationship of the WEE nonstructural region compared to other alphaviruses.  
         [0051]    [0051]FIG. 6. Expression of WEE structural genes in cell culture.  
         [0052]    [0052]FIG. 7. In vitro transcription and translation of WEE expression vectors.  
         [0053]    [0053]FIG. 8. WEE mouse infectivity model.  
         [0054]    [0054]FIG. 9. Protection using ballistic delivery of pCXH-3.  
         [0055]    [0055]FIG. 10. Protection using ballistic delivery of pVHX-6  
         [0056]    [0056]FIG. 11. Protection using ballistic delivery of pVHX-6. 
     
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0057]    The complete nucleotide sequence of the 71V-1658 strain of western equine encephalitis (WEE) virus was determined (minus twenty-five nucleotides from the 5′ end) and shown in SEQ ID NO: 1. A 5′ RACE reaction was used to sequence the 5′ terminus from WEE strain CBA87. The deduced WEE genome was 11,508 nucleotides in length, excluding the 5′ cap nucleotide and 3′ poly(A) tail. The nucleotide composition was 28% A, 25% C, 25% G and 22% U residues. Comparison with partial WEE sequences of strain 5614 (nsP2-nsP3 of the nonstructural region) and strain BFS 1703 (26S structural region) revealed comparatively little variation; a total of 149 nucleotide differences in 8624 bases (1.7% divergence), of which only 28% of these changes (42 nucleotides) altered the encoded amino acids. Comparison of deduced nsP1 and nsP4 amino acid sequences from WEE with the corresponding proteins from eastern equine encephalitis (EEE) yielded identities of 84.9% and 83.8%, respectively. Previously uncharacterized stem loop structures were identified in the nontranslated terminal regions.  
         [0058]    A 3100 bp clone was identified (pcDNA-12) from the 3′ end of the structural genes. A 1500 bp fragment was PCR amplified and cloned into the 5′ end of pcDNA-12 to produce a complete clone of the structural genes (XH-7) as shown in SEQ ID NO: 2. A cDNA clone (pCXH-3) in which the structural genes of WEE strain 71V-1658 were placed under the control of a cytomegalovirus promoter was made, and transfected into tissue culture cells. The viral envelope proteins were functionally expressed in tissue culture, as determined by histochemical staining with monoclonal antibodies which recognize WEE antigens. The construct was used to immunize mice ballistically and intramuscularly. Mice protected ballistically had a significantly reduced risk of infection, against a subsequent intranasal challenge with WEE virus. A new vector was constructed to determine if increased levels of expression could be obtained. The construct used a pVAX vector to express the WEE structural genes (pVHX-6). Upstream portion of the pVHX-6 vector to where it becomes the XH-7 sequence is shown as SEQ ID NO: 3. The remaining nucleotide sequence of pVHX-6 from the point of divergence is identical to that of structural gene pcDWXH-7 of SEQ ID NO: 2.  
         [0059]    Materials and Methods  
         [0060]    Virus Culture and Purification  
         [0061]    Tissue culture was maintained in accordance with established methods (Bird &amp; Forrester, 1981). Minimal essential media containing 5% fetal calf serum (5% DMEM) was used to grow Vero (CRL 1586) and Chinese hamster ovary (CHO) K1 (CCL 61) cells obtained from American Type Culture Collections. A 10% suckling mouse brain (SMB) suspension of WEE strain 71V-1658 was kindly provided by Dr. Nick Karabatsos, Centers for Disease Control, Fort Collins, Colo. WEE Fleming and California strains were purchased from ATCC (Mannanas, Va.). WEE B11 and CBA87 strains were kindly provided by Dr. George Ludwig, United States Army Medical Research Institute of Infectious Disease (Frederick, Md.). Seed stocks of WEE strains were made by inoculation of Vero cells with virus suspensions at a multiplicity of infection (MOI) of less than 0.1. For RNA isolation, virus stocks were prepared by infecting Vero cells at a MOI of 10. The virus was precipitated from cleared supernatant by the addition of polyethylene glycol MW 6000 to 7%(w/v) and NaCl to 2.3%(w/v). It was subsequently purified on a 20-60%(w/w) continuous sucrose gradient, followed by resuspension in PBS.  
         [0062]    Nucleic Acid Preparation  
         [0063]    Viral RNA used in WEE strain 71V-1658 library construction was prepared by the lysis of virus in 0.5%(w/v) sodium dodecyl sulfate (SDS), and RNA extracted using the cesium chloride/guanidium isothiocyanate method previously described (Sambrook et al., 1989). RNA was precipitated using sodium acetate and ethanol, then stored at −70° C. Prior to use, RNA was washed with 80%(v/v) ethanol, dried and dissolved in nuclease-free water (Promega, Madison, Wis.). Integrity of the RNA was checked on formaldehyde agarose gels (Sambrook et al., 1989). A cDNA library of WEE strain 71V-1658 was made by Invitrogen (San Diego, Calif.), by the ligation of cDNA into the BstXI site of prepared pcDNAII vector, and electroporation into electrocompetent DH1 F′  Escherichia coli  cells. Manipulation of RNA and DNA followed established procedures (Sambrook et al, 1989; Ausubel et al., 1995). Rapid plasmid preparations were made using the Wizard™ plasmid purification kit (Promega, Madison, Wis.). Large-scale plasmid preparations used the alkali lysis protocol as modified by Qiagen (Chatsworth, Calif.). For PCR, RT-PCR and DNA sequencing, oligonucleotide primer design was guided by information from WEE strain BFS1703 and other partially sequenced WEE strains (Hahn et al., 1988; Weaver et al., 1993), and from regions of sequence conservation (Ou et al., 1982 &amp;1983). Oligonucleotides were synthesized and gel purified either at the Regional DNA Synthesis Laboratory (Calgary, Alberta), or on a Beckman Oligo 1000 DNA synthesizer. A catalog with the sequences of primers used is listed in Table 1.  
         [0064]    Construction of pCXH-3  
         [0065]    The Invitrogen WEE library was screened by dot blot hybridization (Sambrook et al., 1989) with [ 32 P]-labeled, random primed RT-PCR fragments as probes (Amersham, Oakville, ON). A 3100 bp insert, pcDW-12, was identified, and corresponded to the 3′ end of the 26 S RNA. The missing 5′ end of the 26S region was generated by RT-PCR using the primers WEE5′Sst1 and WEEP3 (Table 1). The 1500 bp SstI/NcoI restricted fragment was inserted into the plasmid, phT3T7BM+(Boehringer Mannheim, Laval, PQ), to generate a XbaI site on the 5′ end. The 1500 bp XbaI/NcoI fragment was excised, gel purified and inserted into the XbaI and NcoI restriction sites of pcDW-12. The resulting clone, pcDWXH-7, encoded the complete 26S region of WEE 71V-1658. The structural gene insert from pcDWXH-7 was cloned into the mammalian expression vector, pCI (Promega, Madison, Wis.). The pcDWXH-7 plasmid was first linearized using HindIII, followed by a Klenow fragment reaction to fill in the 5′ overhang. The insert was then excised using XbaI, gel purified and ligated into the XbaI/SmaI digested pCI vector. The isolated recombinant plasmid, pCXH-3, was characterized as having the correct insert by restriction mapping.  
         [0066]    Construction of pVHX-6  
         [0067]    The clone, pcDWXH-7, encoded the complete 26S region of WEE 71V-1658 was digested with Sac I, and religated in the reverse orientation. The isolate, pcDWHX-45, contained the complete 26S of WEE, with the reverse cloning sites (HindIII on the 5′ end and XbaI on the 3′ end). The WEE 26S gene segment was excised from pcDWHX-45, and cloned into the HindIII and XbaI sites of the mammalian expression vector, pVAX (Invitrogen, La Jolla, Calif.). After transformation into  E. coli  DH10α (Life Sciences, Burlington, ON) and screening of inserts by restriction analysis, a resulting isolate, pVHX-6 was identified. SEQ ID NO: 3 shows the upstream portion of the pVHX-6 vector to where it becomes the XH-7 sequence. The remaining nucleotide sequence of pVHX-6 from the point of divergence is identical to that of structural gene pcDWXH-7 of SEQ ID NO: 2.  
         [0068]    Expression of the Structural Genes of WEE  
         [0069]    The pCXH-3 expression vector was transfected into Vero or CHO K1 cells using the cationic lipid, Lipofectamine™ (Gibco/BRL, Burlington, ON). Briefly, Vero or CHO K1 cells were grown to 30-50% confluency in Costar 6-well plates. The monolayers were transfected with pCXH-3 in accordance with the manufacturer&#39;s directions, for a period of 5 hrs, followed by a further 29 hr incubation after the addition of 5% DMEM. The monolayers were fixed in methanol:acetone (1:1) for 5 min and washed with PBS containing 0.1%(v/v) Tween 20 and 3% BSA (PBS-TB). The cells were incubated 45 min at 37° C. with a {fraction (1/100)} dilution (in PBS-TB) of concentrated cell supernatant from hybridoma cell lines expressing monoclonal antibodies to the WEE E1 (clone 11D2) or E2 (clone 3F3) proteins, followed by washing with PBS-TB. Monolayers were incubated with a {fraction (1/4000)} dilution of goat anti-mouse IgG/IgM (H &amp; L) horse radish peroxidase conjugate (Caltag, So. San Francisco, Calif.) for 45 min at 37° C. After washing with PBS-T, 2 mL of TruBlue™ HRP substrate (Kirkegaard &amp; Perry Laboratories, Gaitherburg, Md.) was added, and plates were incubated a further 30 min at room temperature, followed by microscopic examination.  
         [0070]    In a second method, one-step in vitro transcription and translation reactions using the TNT coupled system (Promega Corporation, Madison, Wis.) was used to express the gene products from both pCXH-3 and pVHX-6, as both have an upstream T7 promoter which can be used for in vitro expression of inserts. The RNA was translated in the presence of [ 35 S]methionine to produce radiolabeled WEE proteins, which were further processed with canine pancreatic microsomal membranes. All components of the in vitro transcription and translation reactions were incubated together for 90 min at 30° C. Results were analyzed by SDS-PAGE or radioimmunoprecipitation.  
         [0071]    Radioimmunoprecipitation  
         [0072]    The TNT reactions were diluted to a volume of 500 ml with RIP buffer consisting of 0.15 M sodium chloride, 0.1% SDS, 50 mM Tris-HCl pH 7.4, and 1% Triton X-100. They were then preabsorbed by incubating with 75 L of protein G-agarose (Gibco BRL) for 30 min at room temperature. The samples were centrifuged at 13,000 rpm for 1 min, and the supernatants were then immunoprecipitated with either 100 μL of supernatants from anti-WEE hybridoma cells or 20 μg of purified anti-WEE antibodies. The reactions were incubated for 1.5 hr at room temperature, after which 75 μL of protein G-agarose was added. The reactions were incubated for an additional 30 min at room temperature. Immunoprecipitated proteins were collected by centrifuging at 13,000 rpm for 1 min. The pellets were washed with 500 μL of RIP buffer and centrifuged at 13,000 rpm for 1 min; this step was repeated three additional times. The pellets were resuspended in 2× Laemmli sample buffer (Bio-Rad Laboratories) containing fresh 2% b-mercaptoethanol and heated at 100° C. for 10 min. The samples were centrifuged at 13,000 rpm for 1 min, and the supernatants were collected. The immunoprecipitated [ 35 S]labeled WEE proteins were further analyzed by SDS-PAGE and autoradiography. Radiolabelled [ 14 C]molecular weight markers from Amersham Pharmacia Biotech were also run on the polyacrylamide gels.  
         [0073]    DNA Sequencing  
         [0074]    Automated sequencing of the 26S region was performed using the ABI Prism Dye Terminator Cycle Sequencing or Big-Dye™ Terminator Cycle Sequencing kits of plasmid templates according to the manufacturer&#39;s instructions (PE-Applied Biosystems, Foster City, Calif.). Sequencing reactions were purified on Centri-Sep™ columns (Princeton Separations, Adelphia, N.J.), dried and analyzed on an ABI 373 or 310 automated sequencer. For the nonstructural region, template cDNAs were generated in a single-step integrated RT-PCR procedure using the Titan™ RT-PCR kit (Boehringer Mannheim, Laval, PQ), following the manufacturer&#39;s suggested protocols. RT-PCR products were purified using the QIAquick™ PCR Purification kit (Qiagen, Chatsworth, Calif.) and sequenced (50-100 ng DNA per reaction). The extreme 5′ end of the genome was not sequenced in WEE 71V-1658. However, a 5′ RACE reaction (Frohman et al., 1988) was used to obtain a cDNA fragment from the 5′ terminus of WEE strain CBA87. Briefly, primer WEE559 (GGTAGATTGATGTCGGTGCATGG) was used to prime reverse transcription of the 5′ terminus of the viral RNA. After poly(A) tailing of the cDNA with terminal transferase, a plus sense primer (GTACTTGACTGACTGTTTTTTTTTTTTTTT) was used in conjunction with WEE559 for amplification of the 5′ terminus.  
         [0075]    Nucleotide Sequence Analysis and Assembly  
         [0076]    Sequence traces were edited manually and assembled using the Seqman component of the Lasergene DNA analysis software (DNASTAR, Madison, Wis.). Codon preferences and patterns were assessed using the CodonUse and CodonFrequency programs, while the overall frequency of mononucleotide and dinucleotides was calculated using the Composition program (Wisconsin Package, Version 9.0, Genetics Computer Group, Madison, Wis.). Quantitative assessments of sequence similarities (nucleotide and amino acid), were calculated by preliminary alignment using the Pileup program, followed by manual alignment adjustment, and analysis with the Distances program (GCG). Amino acid sequences aligned as described, were used as the basis for generating phylogenetic trees (GCG). The GeneQuest module of the Lasergene program (DNASTAR, Madison, Wis.) was used to predict and calculate RNA secondary structures at the ends of the genomic RNA using minimal energy calculations. Multiple sequence alignments were accomplished using the Clustal component of MegAlign (DNASTAR). The complete WEE genomic nucleotide sequence has been submitted to GenBank (Accession Number AF143811).  
         [0077]    Administration of DNA or Inactivated Virus  
         [0078]    DNA solutions or an inactivated WEE virus vaccine in PBS, were administered to the mice by ballistic or intramuscular (IM) routes. For IM route of administration, a 27 g needle was used to deliver 50 μg of DNA (pCXH-3 or pCI-negative control) or 50 μL of inactivated WEE vaccine (SALK WEE inactivated vaccine). The volume of inoculum used was 100 μL, diluted in PBS. Fifty μL was administered IM to each of the hind leg muscles of a mouse. When boosters were given, they were administered 14-28 days apart. For ballistic administration, mice were shaved in the abdominal area with electric hair clippers. The mouse was subjected to ballistic delivery of DNA coated onto gold particles following the manufacturer&#39;s standard specifications. The Helios Gene Gun (Biorad, Mississauga, ON) was used as directed, at a pressure setting of 400 psi. Mice were given 1.25 μg DNA and 0.5 mg gold, 1 μm diameter, per shot, and up to three shots for one dose time. Boosters were given 14-28 days apart. The mice were challenged 14-28 days after the final booster.  
         [0079]    Mouse Infectivity with WEE  
         [0080]    Female BALB/c mice, 17-25 g, were obtained from the mouse breeding colony at Defence Research Establishment Suffield (DRES), with the original breeding pairs purchased from Charles River Canada (St. Constant, Quebec, Canada). The use of these animals was reviewed and approved by Animal Care Committee at DRES. Care and handling of the mice followed guidelines set out by the Canadian Council on Animal Care. Virus was administered to the mice by intranasal (IN) or intraperitoneal (IP) routes. The volumes of inoculum used were 50 μL for IN and 100 μL for IP. For IN administration, mice were anaesthetized with sodium pentobarbital (50 mg/kg body weight, intraperitoneal). When the animals were unconscious, they were carefully supported by hands with their nose up, and the virus suspension in PBS was gently applied with a micropipette into the nostrils. The applied volume was naturally inhaled into the lungs. For IP infection, the mouse was manually restrained, and a 1 ml tuberculin syringe fitted with a 27 g needle was used to administer approximately 100 μL of the virus suspension in PBS. Infected animals were observed daily, for up to 14 days post infection.  
         [0081]    Results  
         [0082]    Complete Nucleotide Sequence of WEE Genome and Deduced Amino Acids  
         [0083]    The nucleotide sequence of WEE strain 71V-1658 (SEQ ID NO: 1) was determined via several distinct sequencing strategies, as summarized in FIG. 1. The 5′ terminus of 25 nt was not determined for this strain. However, it was determined by sequencing a 5′ RACE product from strain CBA87. Excluding the terminal 5′ cap structure and the 3′ poly(A) tail, the genomic sequence of WEE was found to be 11,508 bases long. The base composition was 28% A, 25% C, 25% G, and 22% U. The dinucleotide usage of the WEE genome was compared with those values anticipated from the base composition. Several dinucleotides were found in lower proportions than anticipated, notably UpA (81%), CpG (83%) and CpC (85%) (data not shown). Codons containing the CpG dinucleotide were present at 82% of the anticipated value, including codons for serine (78%), proline (80%) and arginine (78%).  
         [0084]    The WEE 71V-1658 sequence was used to conduct a variety of phylogenetic analyses with previously determined alphavirus sequences. The alphaviruses used in the analyses included EEE strain North American variant (Genbank Ace. No. X671 11), O&#39;Nyong Nyong (ONN) strain Gulu (Genbank Ace. No. M33999), Ross River (RR) strain NB5092 (Genbank Ace. No. M20162), Semliki Forest (SFV) (Genbank Ace. No. J02361), SIN strain HR (Genbank Ace. No. J02363) and VEE ID (Genbank Ace. No. L04653). The degree of conservation among the various sequences (nucleotide and amino acid) through the stereotypical alphavirus genome is shown in Table 2. The carboxy-terminal domain of nsP3, which consistently fails to exhibit homology among sequenced alphaviruses, was excluded from this comparison as it has been adjusted for in previous analysis (Weaver et al., 1993). The deduced amino acid sequences for nsP1-4 of WEE 71V-1658 demonstrated closest identity to the corresponding proteins from EEE (Table 1), reflecting similar observations made for nsP2 and nsP3 of WEE 5614 and EEE (Weaver et al., 1993).  
         [0085]    Nontranslated Terminal Regions  
         [0086]    Alignment of the 5′ terminal nucleotide sequences of WEE CBA87 and WEE 71V-1658 is shown in FIG. 2 a , along with a comparison of the 5′ termini from EEE and VEE. The close similarity between WEE and EEE, has been verified experimentally, in that a EEE/Highlands J degenerate primer, EHJ5′, was able to PCR amplify the 5′ end of the WEE genome, while an analogous SIN primer could not (data not shown).  
         [0087]    Potential stem loop structures were found in WEE 71V-1658, including a stem loop at the extreme 5′ terminus (2-30) and a pair of stem loops (137-189) (FIG. 3 a ). The homologous structures for EEE are also shown (FIG. 3 b ) (Ou et al., 1983). Minimal energy values calculated for the stem loops were similar between WEE and EEE. Further analysis of the region between the structures described above, indicated a large, highly base-paired stem loop structure (39-131), that had not been previously described, and was observed in SIN and EEE in a similar location (data not shown).  
         [0088]    The sequence of WEE 71V-1658 3′ NTR, overall, shared little homology with any of the alphaviruses examined, but included the highly conserved 19 nt region at the 3′ end (11490-11508), which was identical to that determined for WEE BFS1703 by Hahn et al., 1988. Two copies of the characteristic 40 base Sindbis-like terminal repeats as previously reported (On et al., 1982) were found in WEE 71V-1658 (11234-11273 and 11292-11331). However, the 3′ NTR of WEE showed some surprising results that had not been previously described. The first 40 nt terminal repeat formed the backbone for the formation of a 57 nt double stem loop structure (11228-11284) (FIG. 4 b ), consisting of an α and β loop. The second 40 nt repeat of WEE formed a nearly identical 59 nt double stem loop structure (11285-11343), directly adjacent to the first structure. SIN with three 40 nt repeats, forms three double stem loops (FIG. 4 a ) while EEE, which does not contain a SIN-like 40 nt repeat, contains the α and β loops (FIG. 4 c ).  
         [0089]    Nonstructural Region  
         [0090]    Comparisons within the nonstructural regions (4475 nt) of WEE strains 71V-1658 and 5614 (Weaver et al., 1993), yielded 94 nt changes resulting in 26 amino acid substitutions (1.8% difference) as summarized in Table 2. The most notable variation, a three-base deletion (4530) within the nsP3 gene of WEE 71V-1658 constitutes the only insertion/deletion observed within the polypeptide encoding regions. A short hypervariable region was observed (1421-1449), where 11 of 28 nt were different between the two WEE strains (FIG. 2 b ). The presence of an opal termination codon and partial read-through site at the junction of nsP3 and nsP4 is consistent with WEE 5614. Extending previous phylogenetic analyses of WEE (Weaver et al., 1993, 1997), phylogenetic trees depicting viral relatedness were constructed with the Distances program (GCG), for the unexamined genes (nsP1, nsP4) and the entire nonstructural polypeptide encoding region (FIG. 5). The data reveals the close relationship of WEE to EEE, relative to the other alphaviruses analyzed.  
         [0091]    Structural Genes  
         [0092]    The largest WEE cDNA clone isolated, pcDW-12, was 3100 bp in size, but missing 5 nt and the poly(A) tract from the 3′ end as determined by restriction mapping and DNS sequence analysis. The missing 5′ 1500 bp fragment was synthesized using PCR (primers WEE5′Sst1 and WEEP3) and subsequently cloned into pcDW-12 to yield a full-length clone of the structural genes (pcDWXH-7) (SEQ ID NO: 2). Comparison of the structural region of WEE 71V-1658 with WEE BFS 1703 (Hahn et al., 1988), indicated 53 nt changes, resulting in only 11 amino acid differences, of which two were nonconserved. One difference in residue was observed from the amino acid sequence of the N-terminus of the E2 protein of the WEE MacMillan strain (Bell et al., 1983), when this was compared to the deduced protein sequence of 71V-1658. A short fragment (802 nucleotides) of the WEE 71V-1658 E1 protein gene, and the 3′ NTR had been published previously (Weaver et al., 1997); comparison with the sequence reported herein indicated no differences.  
         [0093]    Expression of Structural Gene  
         [0094]    Expression of the insert from the cytomegalovirus (CMV) promoter was accomplished by transfection of the pCXH-3 plasmid into either Vero or CHO K1 cells. Cells expressing the E1 or E2 proteins were detected through the use of specific E1 or E2 monoclonal antibodies to WEE, followed by histochemical staining with the HRP substrate, Tru-Blue as demonstrated in FIG. 6 a.  The control cells transfected with pCI alone showed no staining (FIG. 6 b ), thus, demonstrating the fidelity of the proteins translated from the cloned 26S region. In vitro translation of the insert using TNT T7 rabbit reticulysate and canine microsome system demonstrated synthesis of  35 S-methionine-labelled proteins of the correct size as indicated by immunoprecipitation with monoclonal antibodies to the NC, E1 and E2 proteins (data not shown). Similarly, the construct pVHX-6 was along demonstrated to produce the correct MW proteins as determined by in vitro transcription/translation. The level of expression for pVHX-6 was significantly higher then for pCXH-3 (FIG. 7).  
         [0095]    Protection Against WEE Infection Using DNA Immunization  
         [0096]    Different strains of WEE were shown vary in their virulence in BALB/c mice. When similar amounts of WEE were given intranasally to BALB/c mice, time to death varied from 4 to 8 days. The California and Fleming strains were the most virulent (FIG. 8), and the Fleming strain was chosen as the challenge strain in protection studies. IP administration of the virus did not kill adult mice (data not shown). Intramuscular administration of pCXH-3 did not show any protection, using one or two doses of 50 μg, followed by challenge 30 to 90 days after the final dose (data not shown). Intramuscular administration did result in an increase in antibody titre to WEE as determined by ELISA using a monoclonal antibody to the E1 protein of WEE (data not shown). Expression and protection of pCXH-3 DNA when delivered ballistically. pCI was used as a control DNA. When two doses of pCXH-3 was given, protection of 50% was demonstrated as compared to no protection for pCI (FIG. 9) or PBS controls (data not shown). IM injection showed marginal protection (one group 25% survival-data not shown). The dose of WEE Fleming strain (challenge strain) was 1.25×10 4  PFU for 100% killing via an intranasal route of infection. Preliminary studies examining protection using the pVHX-6 vector, indicated promise with this construct using the Gene Gun, and ballistic delivery. With the pVHX-6 vector, one mouse succumbed immediately to the effects of the sodium pentabarbital (anaesthetic). The remaining three mice showed no signs of coming down with a WEE infection, and remained completely heathy (FIG. 10). Of the four pVAX control mice, all showed signs on WEE infection, and two of the four mice died, while two did recover. A repeat of this experiment using 3 or 4 doses of pVHX-6, given 2 weeks apart, showed complete protection of the mice, similar to 3 doses of WEE inactivated vaccine (FIG. 11). Three or 4 doses of pVAX showed results similar to the saline control, with only about 60% of the mice surviving FIG. 11.  
         [0097]    Discussion  
         [0098]    The WEE 71V-1658 genomic sequence of 11,508 bases was determined directly from cDNA clones of WEE or via sequencing RT-PCR products. The first 25 bases of the WEE genome was determined indirectly, through the use of a 5′ RACE reaction in WEE CBA87. Noting the relatively high conservation in the WEE sequences overall (1.7% divergence) and in the overlap region between the two WEE sequences (see FIG. 2 a ), it appears that the 5′ ends of 71V-1658 and CBA87 are of similar size and sequence.  
         [0099]    Comparison of WEE 71V-1658 to other partial sequences of WEE (Hahn et al., 1988; Weaver et al., 1993) suggests little variation at the nucleotide level among these viruses (Table 2), showing an overall nt sequence difference of 1.7% over 8624 nt. Given a calculated rate of divergence of 0.028% per year for the WEE El protein (Weaver et al., 1997), the expected nt divergence for a difference in isolation of 18 years between the strains, should be 0.5% (71V-1658 isolated in 1971 and BFS 1703 in 1953). The E1 protein itself showed a rate of divergence of 1.5% in nt sequence between 71V-1658 and BFS1703. The lower rate observed by Weaver et al., (1997) could be due to greater conservation of structure at the C terminus of E1, from where the rates of divergence were calculated. Areas with high rates of divergence were observed between WEE strains 71V-1658 and 5614 at the 3′ end of nsP1 and the 5′end of nsP4 (Table 2). The relatively high interstrain value for nsP1 (4.5% difference) may be due to the presence of a small hypervariable region, with 11 of 28 nt changed in strain 5614 (FIG. 2 b ). Variation in nsP4 occurred in a stretch of 21 nt at the 3′ end of the 5614 sequence, and were left out of subsequent homology comparisons (similarity with the EEE sequence was maintained in this region). Discounting the carboxy-terminal region of nsP3 also gives a more accurate picture of the homology of the nsP1-4 nonstructural region (Weaver et al., 1993). The results for comparison of nt and protein sequences of WEE to other alphaviruses is shown in Table 2, and are similar to those obtained with nsP2 and nsP3 of 5614, when compared to other alphavirus sequences. Phylogenetic analysis of the WEE 71V-1658 deduced protein sequences of nsP1, nsP4 and the nsP1-4 region, as related to other alphaviruses (FIG. 5), illustrates the close relationship to EEE (HJ sequences were very limited for comparative purposes and were not included).  
         [0100]    Assessments of codon usage frequencies and the frequency at which certain dinucleotides are found throughout the genome identified a number of statistical anomalies. The slight CpG dinucleotide deficiency previously described within other alphaviruses, and WEE itself, was confirmed in this study, at levels comparable to those reported (Weaver et al., 1993). The CpG under representation is a typical feature of vertebrate genomes, and is not seen in invertebrates. Viruses which infect dual hosts, such as the arboviruses, might be expected to utilize an intermediate nucleotide bias, as indicated by the slight CpG under-utilization observed in alphaviruses (Weaver et al., 1993). A pronounced under-representation of two other dinucleotides was also observed within the WEE genome, UpA, and CpC, a phenomenon noted throughout the genome, though the role of these codon preferences is unclear.  
         [0101]    The 5′ NTR sequence of WEE shows a close phylogenetic affiliation to EEE, and to HJ, although the HJ sequence information is more limited. Ou et al., (1983) had previously predicted (based on minimal free energy calculations) two hairpin structures at the 5′ NTR of several alphaviruses including SIN and EEE. Both structures are present in WEE, the first of which is a 5′ terminal hairpin structure (2-30), similar to that calculated for EEE (FIGS. 3 a  and  b ). The second is a dual hairpin structure (137-162, 165-189) which is almost identical to that identified for EEE. The region between the terminal and dual hairpins can itself form a long hairpin structure, and includes highly conserved stretches of 92 nt (data not shown). The significance of these structures is currently unknown.  
         [0102]    Previous reports (Hahn et al., 1988; Pfeffer et al., 1998) suggested WEE virus arose as a result of two recombination events between alphavirus-like ancestral viruses. The first recombination occurred near the junction of the E3 and capsid genes. The second recombination occurred 80 nucleotides from the 3′ end of the genome. Evidence for the occurrence of the second recombination event is inferred from sequence similarities of the 3′ NTR between WEE, EEE and SIN, in which WEE shows greater similarity to EEE (65%) than to SIN (50%) in the last 100 nt of the 3′ end. However, the apparent plasticity of the 3′ NTR may only be reflecting the selective pressures under which the nascent WEE virus evolved, resulting in rapid selection of 3′ sequences which are more similar to EEE, and may not represent an actual recombination event as previously postulated.  
         [0103]    The 3′ NTRs of alphaviruses are characterized by widespread sequence divergence and yet contain small, strongly conserved motifs (reviewed in Strauss &amp; Strauss, 1994; Pfeffer et al., 1998). Analysis of the 3′ NTR indicated the presence of double stem loop structures among SIN and WEE (FIGS. 4 a  and  b ). Interestingly, the 40 bp repeat found in SIN and WEE is contained within the double stem loop structure. SIN was found to contain 3 double stem loop structures and WEE was found to contain two. In SIN, the spacing between the three double stem loop structures was around 30 nucleotides, while in WEE the distance was zero nt separating the structures. Additional alphaviruses were assessed and it is interesting to note that double stem loop structures were found in many of the WEE- and SIN-related viruses (SIN, Aura, Babanki, Ockelbo, Kyzylagach, Whataroa, WEE and HJ). The double stem loop structures found in SIN and WEE viruses consisted of the α loop (AUGUA[U/C]UU) and the β loop (GCAUAAU) (FIG. 4 b ). Surprisingly, while EEE does not have the 40 bp repeat element found in SIN and WEE, it contains the α and β loop structures (FIG. 4 c ). The significance of these conserved loop structures between SIN, WEE and EEE viruses has yet to be elucidated, although previous studies suggest a role in viral replication and/or host specificity (Kuhn et al., 1990; Kuhn et al., 1991). For example a deletion of 26-318 nt from 3′ end of SIN, resulted in reduced viral replication in mosquito cells but not in chicken cells (Kuhn et al., 1990). In contrast, substitution of the SIN 3′ NTR with the substantially different RR 3′ NTR (which lacks the 40 bp repeat and double stem loop structures), had no effect on the growth of the chimeric virus in mosquito cells, suggesting that host proteins interact with the 3′ NTRs to cause differential host effects (Kuhn et al., 1991).  
         [0104]    The 26S region of 71V-1658 was placed under the control of the CMV promoter of pCI. To test for functional expression of the pCXH-3 vector and for a functional product in cell culture, the pCXH-3 vector was transiently transfected into Vero cells. WEE proteins were detected on the cell using specific monoclonal antibodies to both the E1 (FIG. 6 a ) and E2 proteins (data not shown). The binding specificity of these monoclonals has been previously determined by western blot analysis and immunoprecipitation analysis (data not shown). The use of pCXH-3 in DNA immunization experiments indicated that the construct could partially protect against WEE intranasal challenge using ballistic delivery. Preliminary results do indicate that WEE reactive antibodies can be detected by ELISA when the pCXH-3 plasmid is given intramuscularly (unpublished results). However, this afforded no protection to the mice, as there were no survivors. Intranasal (data not shown) delivery of the pCXH-3, with and without liposome encapsulation did not demonstrate any protection under the conditions used. Mice immunized with the pCI control plasmid did not show any signs of protection in these studies.  
         [0105]    Expression of the WEE structural proteins in the pCI-based vector (pCHX-3) gave moderate to poor levels of expression in vitro, using the TNT expression kit. A new vector, pVAX (Invitrogen) was designed for DNA immunization and was basically the same as pCI, but lacked the intron found in the pCI vector. Initial restriction mapping of pCXH-3 indicated the plasmid was the expected size, but later analysis indicated a extra 4 kb fragment was present (data not shown). The WEE structural proteins were cloned and expressed in pVHX-6, indicating the correct sized proteins by SDS-PAGE, and producing higher levels of WEE product in vitro (FIG. 7). Preliminary results with pVHX-6 indicated it could completely protect mice against an intranasal challenge of WEE. While 50% of the pVAX mice did survive, they all demonstrated at least moderate to severe infection with WEE. It is possible that pVAX contains CpG motifs that show some protective effect, through a nonspecific adjuvant like effect (Kreig et al, 1998). However, there was a dramatic difference between the pVAX and the pVHX-6 group, in the protection afforded the two groups of mice.  
         [0106]    The plasmids, pCXH-3 and pVHX-6 show promise as vaccine candidates for WEE. This is especially important for protection against an aerosol challenge of WEE, and event that would be envisioned in a potential biological warfare attack using WEE as a biological warfare agent. This agent is difficult to protect against if delivered aerosolly, as the agent is purported to travel up the nerves directly into the brain. The research is applicable to VEE and EEE, as these viruses can also cause encephalitis following a similar route of infection (equines and potentially human).  
         [0107]    It is to be understood that the embodiments and variations shown and described herein are merely illustrative of the principles of this invention and that various modifications may be implemented by those skilled in the art without departing from the scope and spirit of the invention.  
                             TABLE 1                           WEE 26S Region Primers            Name   Length   Sequence               WEEPRO   30   AATCACCCTCTACGGCTGACCTAAATAGGT               WEEPR-SST   24   GGCTGAGCTCAATAGGTGACGTAG               WEE3′   30   GTAGTGTATATTAGAGACCCATAGTGAGTC               WEE5′SST   20   TCCAGATACGAGCTCATACT               WEEN1   20   GGTGCCGCTGGAGGCCGTTT               WEEN1A   20   GATCTTAGGAGGTCGATAGC               WEEN2   20   GGCTGATGAAACCACTCCAC               WEEN3   20   CCACCCGTGTGCTATTCACT               WEEN3A   20   CGCCGTGTTTCAGCCCAATA               WEEN4   20   TCACGAGCGGAGCATCTGAG               WEEN5   20   GGCATCACCCTCCACCTGAC               WEEN6   20   TTGTTATTCTGTTCCGCTGC               WEEN7   20   CTATTGATCATGCAGTCGCA               WEEN8   20   AGTGGAGCCTCTGCGAGCGT               WEEN9   20   GAGGAGTGGGCGGGAAAGGC               WEEN10   20   CTAAAACTCGATGTATTTCC               WEEN11   20   ACGCGAACGAAGATGAACGG               WEEN12   20   ACTGTCATTGTGCTGTGTGG               WEEN13   20   CACAGTCATTCCTTCACCAC               WEEN14   20   CGTCATCAGAAAGGGGCTTG               WEEN15   20   CAAAGCTGACAGGGAGGGAC               WEEN16   20   GGAAAGCTGGTAAAGTGCCA               WEEN0   20   GGAGAACCACATAAAGTCGA               WNSP1   25   GGCTAACGTGGACAGGGACGTGATG               WEEP0   20   GGCTATCGACCTCCTAAGAT               WEEP0A   20   CTGTCGGTTCCCTGGTTTAG               WEEP1   20   CTGGGGAACGTCGCCATACT               WEEP2   20   CGTTCTCCAGCAGCGTGTCG               WEEP2A   20   TATTGGGCTGAAACACGGCG               WEEP3   20   CTTCAAGTGATCGTAAACGT               WEEP4   20   ACTCCAGCCCTTCTCGCCCC               WEEP5   20   GTTCGACCAACGCCTTATAC               WEEP6   20   AAGGGTGAAAAAGCGGCTGA               WEEP7   20   GGTGATTCTGATGATCTCAC               WEEP8   20   TGGAAACTGCCGCCTGGAAT               WEEP10   20   CCTTGATGTCATGGTCGTGG               WEEP11   20   TGCACTGAGTGGTCTGTGTG               WEEP12   20   ATGTTTCAGCGTTGGTTGGC               WEEP13   20   GTGTTCTCACTGTCACAGAA               WEEP14   20   ATGTGTGGTCGCTTCCTTCA                  
 
         [0108]    [0108]                                                                                           TABLE 2                           Percentage Variation in Nucleotide and Encoded Amino       Acid Sequences Between WEE 71V-1658 and Other Alphaviruses                WEE   WEE                                   (BFS1703)   (5614)   EEE   VEE   SIN   RR   ONN   SF                        5′NTR   —   —                               nsP1 (nt)   —   (4.5)   25.1   34.8   40.9   37.8   39.7   39.1       nsP1 (aa)   —   (6.3)   15.1   32.1   40.3   35.5   37.2   33.3       nsP2 (nt)   —   1.8   28.2   34.6   43.9   42.1   42.9   42.8       nsP2 (aa)   —   0.6   16.2   26.5   44.9   43.2   44.9   44.4       nsP3 (nt)*   —   1.8   30.2   36.7   45.8   39.3   42.6   42.2       nsP3 (aa)*   —   2.1   18.8   32.4   46.3   38.7   40.9   43.5       nsP4 (nt)   (1.8)   (2.4)   25.6   31.4   34.7   35.3   36.0   37.0       nsP4 (aa)   (2.6)   (4.3)   11.7   21.4   26.8   27.3   25.8   27.4       intervening (nt)   4.3   —   56.6   51.5   47.6   44.7   60.0   47.7       Capsid (nt)   2.1   —   26.3   40.8   47.7   46.3   47.5   48.2       Capsid (aa)   1.5   —   16.8   43.5   52.8   53.3   54.6   54.3       E3 (nt)   1.1   —   45.6   40.7   38.3   51.74   7.5   46.7       E3 (aa)   1.7   —   38.0   39.6   39.4   46.0   45.8   43.9       E2 (nt)   1.2   —   51.2   52.3   36.2   51.7   55.3   52.8       E2 (aa)   1.0   —   59.0   60.0   31.7   63.5   65.7   64.7       6K (nt)   0.6   —   53.3   46.3   26.1   51.9   50.3   54.3       6K (aa)   1.8   —   65.6   59.3   32.7   72.2   69.1   75.9       E1 (nt)   1.5   —   43.8   45.8   29.6   47.2   48.5   44.4       E1 (aa)   0.5   —   49.0   51.0   23.4   51.5   54.8   50.3       3′NTR (nt)   0.7   —   57.8   55.0   53.2   69.1   65.8   60.3                                            
         [0109]    [0109] 
     
       
       
         0 
         
               
             
           
               
                   
               
               
                   
               
               
                   
               
               
                                                SEQUENCE LISTING  
               
               
                   
               
               
                   
               
               
                 &lt;160&gt; NUMBER OF SEQ ID NOS: 7  
               
               
                   
               
               
                 &lt;210&gt; SEQ ID NO 1  
               
               
                 &lt;211&gt; LENGTH: 11484  
               
               
                 &lt;212&gt; TYPE: DNA  
               
               
                 &lt;213&gt; ORGANISM: Western equine encephalomyelitis virus - strain 71V-1658  
               
               
                 &lt;220&gt; FEATURE:  
               
               
                 &lt;221&gt; NAME/KEY: CDS  
               
               
                 &lt;222&gt; LOCATION: (25)..(7428)  
               
               
                 &lt;223&gt; OTHER INFORMATION: 5′ UTR &lt;1 .. 24  
               
               
                 &lt;220&gt; FEATURE:  
               
               
                 &lt;221&gt; NAME/KEY: CDS  
               
               
                 &lt;222&gt; LOCATION: (7473)..(11183)  
               
               
                 &lt;223&gt; OTHER INFORMATION: encodes nucleocapsid, E3, E2, 6K and E1 proteins  
               
               
                   
               
               
                 &lt;400&gt; SEQUENCE: 1  
               
               
                   
               
               
                 accctacaaa ctaatcgatc caat atg gaa aga att cac gtt gac tta gat        51  
               
               
                                            Met Glu Arg Ile His Val Asp Leu Asp  
               
               
                                            1               5  
               
               
                   
               
               
                 gct gac agc ccg tat gtc aag tcg tta cag cgg acg ttt cca caa ttt       99  
               
               
                 Ala Asp Ser Pro Tyr Val Lys Ser Leu Gln Arg Thr Phe Pro Gln Phe  
               
               
                 10                  15                  20                  25  
               
               
                   
               
               
                 gag atc gaa gca agg cag gtc act gac aat gac cat gcc aat gcc aga      147  
               
               
                 Glu Ile Glu Ala Arg Gln Val Thr Asp Asn Asp His Ala Asn Ala Arg  
               
               
                                 30                  35                  40  
               
               
                   
               
               
                 gcg ttt tcg cat gtg gca aca aag ctc att gag agc gaa gtc gac cgg      195  
               
               
                 Ala Phe Ser His Val Ala Thr Lys Leu Ile Glu Ser Glu Val Asp Arg  
               
               
                             45                  50                  55  
               
               
                   
               
               
                 gac caa gtt atc ttg gac att gga agt gcg ccc gtc aga cat gca cat      243  
               
               
                 Asp Gln Val Ile Leu Asp Ile Gly Ser Ala Pro Val Arg His Ala His  
               
               
                         60                  65                  70  
               
               
                   
               
               
                 tcc aat cac cgc tat cat tgt atc tgc cct atg ata agc gct gaa gac      291  
               
               
                 Ser Asn His Arg Tyr His Cys Ile Cys Pro Met Ile Ser Ala Glu Asp  
               
               
                     75                  80                  85  
               
               
                   
               
               
                 ccg gac aga cta caa cgg tat gca gaa aga ctt aag aaa agt gac att      339  
               
               
                 Pro Asp Arg Leu Gln Arg Tyr Ala Glu Arg Leu Lys Lys Ser Asp Ile  
               
               
                 90                  95                  100                 105  
               
               
                   
               
               
                 acc gac aag aac ata gcc tct aag gcg gca gac ctg ctg gaa gtc atg      387  
               
               
                 Thr Asp Lys Asn Ile Ala Ser Lys Ala Ala Asp Leu Leu Glu Val Met  
               
               
                                 110                 115                 120  
               
               
                   
               
               
                 tca aca cca gac gca gag act cca tct ctg tgt atg cac aca gac gcc      435  
               
               
                 Ser Thr Pro Asp Ala Glu Thr Pro Ser Leu Cys Met His Thr Asp Ala  
               
               
                             125                 130                 135  
               
               
                   
               
               
                 acg tgt agg tac ttt gga agt gta gca gta tac caa gat gtg tac gca      483  
               
               
                 Thr Cys Arg Tyr Phe Gly Ser Val Ala Val Tyr Gln Asp Val Tyr Ala  
               
               
                         140                 145                 150  
               
               
                   
               
               
                 gtc cat gca ccg aca tca atc tac cac cag gcg ctt aaa gga gtt agg      531  
               
               
                 Val His Ala Pro Thr Ser Ile Tyr His Gln Ala Leu Lys Gly Val Arg  
               
               
                     155                 160                 165  
               
               
                   
               
               
                 aca att tac tgg ata ggc ttt gac acg acc cct ttt atg tac aaa aac      579  
               
               
                 Thr Ile Tyr Trp Ile Gly Phe Asp Thr Thr Pro Phe Met Tyr Lys Asn  
               
               
                 170                 175                 180                 185  
               
               
                   
               
               
                 atg gca ggt tcc tac cct act tac aac acg aac tgg gct gac gag aga      627  
               
               
                 Met Ala Gly Ser Tyr Pro Thr Tyr Asn Thr Asn Trp Ala Asp Glu Arg  
               
               
                                 190                 195                 200  
               
               
                   
               
               
                 gta ttg gaa gca cgt aac att ggc ctc ggt aac tca gat ctt cag gag      675  
               
               
                 Val Leu Glu Ala Arg Asn Ile Gly Leu Gly Asn Ser Asp Leu Gln Glu  
               
               
                             205                 210                 215  
               
               
                   
               
               
                 agc agg ctt gga aaa ctc tca atc ctt agg aag aag agg ctc caa cct      723  
               
               
                 Ser Arg Leu Gly Lys Leu Ser Ile Leu Arg Lys Lys Arg Leu Gln Pro  
               
               
                         220                 225                 230  
               
               
                   
               
               
                 act aat aag atc ata ttc tcg gtt ggt tca aca atc tac aca gaa gat      771  
               
               
                 Thr Asn Lys Ile Ile Phe Ser Val Gly Ser Thr Ile Tyr Thr Glu Asp  
               
               
                     235                 240                 245  
               
               
                   
               
               
                 aga tca ctg tta cgt agc tgg cat ctt cca aac gtg ttc cac ttg aaa      819  
               
               
                 Arg Ser Leu Leu Arg Ser Trp His Leu Pro Asn Val Phe His Leu Lys  
               
               
                 250                 255                 260                 265  
               
               
                   
               
               
                 gga aag tct aac ttc aca ggt aga tgt ggg acc att gtc agc tgt gaa      867  
               
               
                 Gly Lys Ser Asn Phe Thr Gly Arg Cys Gly Thr Ile Val Ser Cys Glu  
               
               
                                 270                 275                 280  
               
               
                   
               
               
                 ggg tac gtc atc aaa aag ata acg atc agc cca gga cta tac ggt aaa      915  
               
               
                 Gly Tyr Val Ile Lys Lys Ile Thr Ile Ser Pro Gly Leu Tyr Gly Lys  
               
               
                             285                 290                 295  
               
               
                   
               
               
                 gtt gag aac ttg gcg tcc aca atg cat cgc gag ggt ttc ttg agt tgc      963  
               
               
                 Val Glu Asn Leu Ala Ser Thr Met His Arg Glu Gly Phe Leu Ser Cys  
               
               
                         300                 305                 310  
               
               
                   
               
               
                 aaa gtc aca gat acg ctg cgc ggc gag agg gtt tct ttt gct gtg tgt     1011  
               
               
                 Lys Val Thr Asp Thr Leu Arg Gly Glu Arg Val Ser Phe Ala Val Cys  
               
               
                     315                 320                 325  
               
               
                   
               
               
                 acg tat gta cca gcc aca ctt tgc gat cag atg aca ggg att ctg gca     1059  
               
               
                 Thr Tyr Val Pro Ala Thr Leu Cys Asp Gln Met Thr Gly Ile Leu Ala  
               
               
                 330                 335                 340                 345  
               
               
                   
               
               
                 act gac gtt agt gtg gat gac gca caa aaa cta ttg gtt ggg ctc aac     1107  
               
               
                 Thr Asp Val Ser Val Asp Asp Ala Gln Lys Leu Leu Val Gly Leu Asn  
               
               
                                 350                 355                 360  
               
               
                   
               
               
                 caa agg att gtc gtc aat ggt agg acg caa aga aat act aac aca atg     1155  
               
               
                 Gln Arg Ile Val Val Asn Gly Arg Thr Gln Arg Asn Thr Asn Thr Met  
               
               
                             365                 370                 375  
               
               
                   
               
               
                 cag aac tat cta tta cca gtg gtc gcc cag gcg ttt tcc agg tgg gcg     1203  
               
               
                 Gln Asn Tyr Leu Leu Pro Val Val Ala Gln Ala Phe Ser Arg Trp Ala  
               
               
                         380                 385                 390  
               
               
                   
               
               
                 cgt gaa cat cgt gcc gac ttg gac gac gag aaa gaa cta ggg gtg cgg     1251  
               
               
                 Arg Glu His Arg Ala Asp Leu Asp Asp Glu Lys Glu Leu Gly Val Arg  
               
               
                     395                 400                 405  
               
               
                   
               
               
                 gag cgc act ctt act atg ggc tgc tgc tgg gct ttc aag acc cag aaa     1299  
               
               
                 Glu Arg Thr Leu Thr Met Gly Cys Cys Trp Ala Phe Lys Thr Gln Lys  
               
               
                 410                 415                 420                 425  
               
               
                   
               
               
                 atc aca tcc atc tac aag aag cct ggt acg caa aca att aag aaa gta     1347  
               
               
                 Ile Thr Ser Ile Tyr Lys Lys Pro Gly Thr Gln Thr Ile Lys Lys Val  
               
               
                                 430                 435                 440  
               
               
                   
               
               
                 cct gcc gtc ttt gac tca ttt gtg att cca cgc ctt acc agc cac ggg     1395  
               
               
                 Pro Ala Val Phe Asp Ser Phe Val Ile Pro Arg Leu Thr Ser His Gly  
               
               
                             445                 450                 455  
               
               
                   
               
               
                 ctc gat atg ggc ttc cgc cgt agg ctc aag ctg ctg ctt gaa cca act     1443  
               
               
                 Leu Asp Met Gly Phe Arg Arg Arg Leu Lys Leu Leu Leu Glu Pro Thr  
               
               
                         460                 465                 470  
               
               
                   
               
               
                 gtc aaa ccc gca ccg gct att aca atg gcc gat gtg gag cat ctg cgt     1491  
               
               
                 Val Lys Pro Ala Pro Ala Ile Thr Met Ala Asp Val Glu His Leu Arg  
               
               
                     475                 480                 485  
               
               
                   
               
               
                 ggc tta cag caa gaa gct gaa gaa gtg gct gca gcg gaa gag atc aga     1539  
               
               
                 Gly Leu Gln Gln Glu Ala Glu Glu Val Ala Ala Ala Glu Glu Ile Arg  
               
               
                 490                 495                 500                 505  
               
               
                   
               
               
                 gaa gcc ctg cca ccc ttg ctc cct gaa ata gaa aaa gag acc gta gag     1587  
               
               
                 Glu Ala Leu Pro Pro Leu Leu Pro Glu Ile Glu Lys Glu Thr Val Glu  
               
               
                                 510                 515                 520  
               
               
                   
               
               
                 gca gaa gta gac ctc att atg caa gag gca gga gca ggt agc gtg gag     1635  
               
               
                 Ala Glu Val Asp Leu Ile Met Gln Glu Ala Gly Ala Gly Ser Val Glu  
               
               
                             525                 530                 535  
               
               
                   
               
               
                 aca cca cga gga cac atc agg gtg aca agt tac cca ggc gaa gag aag     1683  
               
               
                 Thr Pro Arg Gly His Ile Arg Val Thr Ser Tyr Pro Gly Glu Glu Lys  
               
               
                         540                 545                 550  
               
               
                   
               
               
                 att ggg tct tac gct ata ctt tca ccc cag gcg gta ttg aat agt gaa     1731  
               
               
                 Ile Gly Ser Tyr Ala Ile Leu Ser Pro Gln Ala Val Leu Asn Ser Glu  
               
               
                     555                 560                 565  
               
               
                   
               
               
                 aaa ctg gcg tgt atc cac cca ttg gcg gaa caa gta ctg gta atg act     1779  
               
               
                 Lys Leu Ala Cys Ile His Pro Leu Ala Glu Gln Val Leu Val Met Thr  
               
               
                 570                 575                 580                 585  
               
               
                   
               
               
                 cac aaa ggt agg gca ggg aga tac aaa gtc gag cca tac cac ggt aag     1827  
               
               
                 His Lys Gly Arg Ala Gly Arg Tyr Lys Val Glu Pro Tyr His Gly Lys  
               
               
                                 590                 595                 600  
               
               
                   
               
               
                 gtc att gta cca gaa ggg acg gcg gtc cct gtt caa gac ttc cag gca     1875  
               
               
                 Val Ile Val Pro Glu Gly Thr Ala Val Pro Val Gln Asp Phe Gln Ala  
               
               
                             605                 610                 615  
               
               
                   
               
               
                 ttg agt gag agc gct acg atc gtt ttc aac gag agg gag ttc gta aac     1923  
               
               
                 Leu Ser Glu Ser Ala Thr Ile Val Phe Asn Glu Arg Glu Phe Val Asn  
               
               
                         620                 625                 630  
               
               
                   
               
               
                 aga tac ctg cac cac atc gca atc aac gga gga gcg cta aac act gac     1971  
               
               
                 Arg Tyr Leu His His Ile Ala Ile Asn Gly Gly Ala Leu Asn Thr Asp  
               
               
                     635                 640                 645  
               
               
                   
               
               
                 gaa gag tac tat aag act gta aag act cag gac aca gac tca gaa tac     2019  
               
               
                 Glu Glu Tyr Tyr Lys Thr Val Lys Thr Gln Asp Thr Asp Ser Glu Tyr  
               
               
                 650                 655                 660                 665  
               
               
                   
               
               
                 gtc ttc gat att gac gca cga aag tgt gtt aag cga gaa gac gca ggt     2067  
               
               
                 Val Phe Asp Ile Asp Ala Arg Lys Cys Val Lys Arg Glu Asp Ala Gly  
               
               
                                 670                 675                 680  
               
               
                   
               
               
                 ccc ttg tgc cta acc ggt gat ctg gta gat cca cca ttt cac gag ttt     2115  
               
               
                 Pro Leu Cys Leu Thr Gly Asp Leu Val Asp Pro Pro Phe His Glu Phe  
               
               
                             685                 690                 695  
               
               
                   
               
               
                 gcg tac gag agt ctc aag aca cga cca gca gca cct cac aaa gtc cca     2163  
               
               
                 Ala Tyr Glu Ser Leu Lys Thr Arg Pro Ala Ala Pro His Lys Val Pro  
               
               
                         700                 705                 710  
               
               
                   
               
               
                 acc atc gga gtc tat gga gtg cca ggt tca ggt aaa tct gga atc atc     2211  
               
               
                 Thr Ile Gly Val Tyr Gly Val Pro Gly Ser Gly Lys Ser Gly Ile Ile  
               
               
                     715                 720                 725  
               
               
                   
               
               
                 aaa agc gct gtg act aag aaa gat ctg gtt gtg agt gcg aag aag gaa     2259  
               
               
                 Lys Ser Ala Val Thr Lys Lys Asp Leu Val Val Ser Ala Lys Lys Glu  
               
               
                 730                 735                 740                 745  
               
               
                   
               
               
                 aac tgc gca gaa atc atc agg gat gta agg agg atg aga cgt atg gat     2307  
               
               
                 Asn Cys Ala Glu Ile Ile Arg Asp Val Arg Arg Met Arg Arg Met Asp  
               
               
                                 750                 755                 760  
               
               
                   
               
               
                 gtt gct gct agg act gtc gat tca gtg ctt cta aat ggg gtt aag cac     2355  
               
               
                 Val Ala Ala Arg Thr Val Asp Ser Val Leu Leu Asn Gly Val Lys His  
               
               
                             765                 770                 775  
               
               
                   
               
               
                 ccc gtt aac act ctg tac att gat gag gca ttt gcc tgc cat gca ggg     2403  
               
               
                 Pro Val Asn Thr Leu Tyr Ile Asp Glu Ala Phe Ala Cys His Ala Gly  
               
               
                         780                 785                 790  
               
               
                   
               
               
                 acg ctg ctg gca ctg att gcc atc gtc aaa cct aag aaa gtg gta ttg     2451  
               
               
                 Thr Leu Leu Ala Leu Ile Ala Ile Val Lys Pro Lys Lys Val Val Leu  
               
               
                     795                 800                 805  
               
               
                   
               
               
                 tgc ggg gac cca aaa caa tgc ggc ttc ttt aac atg atg tgc ctg aaa     2499  
               
               
                 Cys Gly Asp Pro Lys Gln Cys Gly Phe Phe Asn Met Met Cys Leu Lys  
               
               
                 810                 815                 820                 825  
               
               
                   
               
               
                 gta cat ttt aac cat gac ata tgc act gaa gtg tac cat aaa agc atc     2547  
               
               
                 Val His Phe Asn His Asp Ile Cys Thr Glu Val Tyr His Lys Ser Ile  
               
               
                                 830                 835                 840  
               
               
                   
               
               
                 tct agg agg tgc aca cag act gta acc gcc atc gtc tcc acg ctc ttc     2595  
               
               
                 Ser Arg Arg Cys Thr Gln Thr Val Thr Ala Ile Val Ser Thr Leu Phe  
               
               
                             845                 850                 855  
               
               
                   
               
               
                 tac gac aag cga atg aag acg gtt aac cca tgt gct gat aaa atc atc     2643  
               
               
                 Tyr Asp Lys Arg Met Lys Thr Val Asn Pro Cys Ala Asp Lys Ile Ile  
               
               
                         860                 865                 870  
               
               
                   
               
               
                 ata gat acc aca ggg acc aca aag ccg cac aaa gat gat ctg att cta     2691  
               
               
                 Ile Asp Thr Thr Gly Thr Thr Lys Pro His Lys Asp Asp Leu Ile Leu  
               
               
                     875                 880                 885  
               
               
                   
               
               
                 acc tgt ttc aga gga tgg gtg aaa cag cta cag att gac tac aaa aat     2739  
               
               
                 Thr Cys Phe Arg Gly Trp Val Lys Gln Leu Gln Ile Asp Tyr Lys Asn  
               
               
                 890                 895                 900                 905  
               
               
                   
               
               
                 cac gaa atc atg act gcg gct gca tcg caa gga ctt acg cgg aaa ggc     2787  
               
               
                 His Glu Ile Met Thr Ala Ala Ala Ser Gln Gly Leu Thr Arg Lys Gly  
               
               
                                 910                 915                 920  
               
               
                   
               
               
                 gtt tat gct gtc agg tac aaa gtc aac gag aat cca ctc tac tcg cag     2835  
               
               
                 Val Tyr Ala Val Arg Tyr Lys Val Asn Glu Asn Pro Leu Tyr Ser Gln  
               
               
                             925                 930                 935  
               
               
                   
               
               
                 act tct gag cac gtg aac gtg tta ctt aca cgc aca gaa aaa cgc att     2883  
               
               
                 Thr Ser Glu His Val Asn Val Leu Leu Thr Arg Thr Glu Lys Arg Ile  
               
               
                         940                 945                 950  
               
               
                   
               
               
                 gtc tgg aag acg cta gct ggt gat ccc tgg ata aag aca ctt aca gct     2931  
               
               
                 Val Trp Lys Thr Leu Ala Gly Asp Pro Trp Ile Lys Thr Leu Thr Ala  
               
               
                     955                 960                 965  
               
               
                   
               
               
                 aaa tat ccc ggg gat ttc acg gct tca ttg gac gac tgg cag cgc gaa     2979  
               
               
                 Lys Tyr Pro Gly Asp Phe Thr Ala Ser Leu Asp Asp Trp Gln Arg Glu  
               
               
                 970                 975                 980                 985  
               
               
                   
               
               
                 cac gac gcc att atg gca cgc gtt ctt gat aag ccg cag aca gct  gat    3027  
               
               
                 His Asp Ala Ile Met Ala Arg Val Leu Asp Lys Pro Gln Thr Ala  Asp  
               
               
                                 990                 995                 1000  
               
               
                   
               
               
                 gtg ttc cag aat  aag gtg aac gtc tgc  tgg gcg aag gct tta  gag      3072  
               
               
                 Val Phe Gln Asn  Lys Val Asn Val Cys  Trp Ala Lys Ala Leu  Glu  
               
               
                             1005                 1010                 1015  
               
               
                   
               
               
                 cca gtc ttg gcc  acg gcc aac att gtg  ctg acg aga cag cag  tgg      3117  
               
               
                 Pro Val Leu Ala  Thr Ala Asn Ile Val  Leu Thr Arg Gln Gln  Trp  
               
               
                             1020                 1025                 1030  
               
               
                   
               
               
                 gag acg ttg cac  cca ttc aag cat gac  aga gcg tac tca cct  gaa      3162  
               
               
                 Glu Thr Leu His  Pro Phe Lys His Asp  Arg Ala Tyr Ser Pro  Glu  
               
               
                             1035                 1040                 1045  
               
               
                   
               
               
                 atg gca ctg aac  ttc ttt tgc acc agg  ttc ttt gga gta gac  ctg      3207  
               
               
                 Met Ala Leu Asn  Phe Phe Cys Thr Arg  Phe Phe Gly Val Asp  Leu  
               
               
                             1050                 1055                 1060  
               
               
                   
               
               
                 gac agt ggg tta  ttt tcc gct cct acc  gtc gca ctt act tac  agg      3252  
               
               
                 Asp Ser Gly Leu  Phe Ser Ala Pro Thr  Val Ala Leu Thr Tyr  Arg  
               
               
                             1065                 1070                 1075  
               
               
                   
               
               
                 gat cag cac tgg  gat aac tcg cca ggg  aag aac atg tat ggg  ctt      3297  
               
               
                 Asp Gln His Trp  Asp Asn Ser Pro Gly  Lys Asn Met Tyr Gly  Leu  
               
               
                             1080                 1085                 1090  
               
               
                   
               
               
                 aat aga gag gta  gca aag gag ttg tca  cgg cga tat ccg tgc  atc      3342  
               
               
                 Asn Arg Glu Val  Ala Lys Glu Leu Ser  Arg Arg Tyr Pro Cys  Ile  
               
               
                             1095                 1100                 1105  
               
               
                   
               
               
                 aca aaa gcg gtt  gac aca ggc agg gta  gct gat ata agg aat  aat      3387  
               
               
                 Thr Lys Ala Val  Asp Thr Gly Arg Val  Ala Asp Ile Arg Asn  Asn  
               
               
                             1110                 1115                 1120  
               
               
                   
               
               
                 acc atc aag gac  tac tct cca aca att  aat gtg gtt cca tta  aat      3432  
               
               
                 Thr Ile Lys Asp  Tyr Ser Pro Thr Ile  Asn Val Val Pro Leu  Asn  
               
               
                             1125                 1130                 1135  
               
               
                   
               
               
                 cgc cgg ttg ccc  cac tcg ttg atc gtt  gac cac aaa gga cag  ggt      3477  
               
               
                 Arg Arg Leu Pro  His Ser Leu Ile Val  Asp His Lys Gly Gln  Gly  
               
               
                             1140                 1145                 1150  
               
               
                   
               
               
                 aca act gat cac  agc gga ttc cta tct  aag atg aag ggc aaa  tct      3522  
               
               
                 Thr Thr Asp His  Ser Gly Phe Leu Ser  Lys Met Lys Gly Lys  Ser  
               
               
                             1155                 1160                 1165  
               
               
                   
               
               
                 gtg ttg gtg atc  ggc gat cct atc agc  att cca ggg aag aaa  gta      3567  
               
               
                 Val Leu Val Ile  Gly Asp Pro Ile Ser  Ile Pro Gly Lys Lys  Val  
               
               
                             1170                 1175                 1180  
               
               
                   
               
               
                 gag tcc atg ggt  cca ttg ccc act aat  acc atc agg tgt gat  ctc      3612  
               
               
                 Glu Ser Met Gly  Pro Leu Pro Thr Asn  Thr Ile Arg Cys Asp  Leu  
               
               
                             1185                 1190                 1195  
               
               
                   
               
               
                 gat ttg gga ata  cct agc cat gtc ggt  aaa tat gac att atc  ttt      3657  
               
               
                 Asp Leu Gly Ile  Pro Ser His Val Gly  Lys Tyr Asp Ile Ile  Phe  
               
               
                             1200                 1205                 1210  
               
               
                   
               
               
                 gtc aat gtt agg  acc ccg tac agg aac  cat cac tac caa cag  tgc      3702  
               
               
                 Val Asn Val Arg  Thr Pro Tyr Arg Asn  His His Tyr Gln Gln  Cys  
               
               
                             1215                 1220                 1225  
               
               
                   
               
               
                 gag gat cac gct  atc cac cac agc atg  cta acg tgt aag gct  gtc      3747  
               
               
                 Glu Asp His Ala  Ile His His Ser Met  Leu Thr Cys Lys Ala  Val  
               
               
                             1230                 1235                 1240  
               
               
                   
               
               
                 cac cac ctg aac  act ggc gga aca tgt  gtg gct ata ggg tat  ggg      3792  
               
               
                 His His Leu Asn  Thr Gly Gly Thr Cys  Val Ala Ile Gly Tyr  Gly  
               
               
                             1245                 1250                 1255  
               
               
                   
               
               
                 ctt gct gat cgc  gca acc gag aat atc  atc act gcg gtg gca  cgc      3837  
               
               
                 Leu Ala Asp Arg  Ala Thr Glu Asn Ile  Ile Thr Ala Val Ala  Arg  
               
               
                             1260                 1265                 1270  
               
               
                   
               
               
                 tca ttt agg ttt  acc cgt gtc tgt cag  cct aag aac act gcc  gaa      3882  
               
               
                 Ser Phe Arg Phe  Thr Arg Val Cys Gln  Pro Lys Asn Thr Ala  Glu  
               
               
                             1275                 1280                 1285  
               
               
                   
               
               
                 aat act gag gtt  ctc ttc gtg ttc ttc  ggc aag gac aac ggc  aac      3927  
               
               
                 Asn Thr Glu Val  Leu Phe Val Phe Phe  Gly Lys Asp Asn Gly  Asn  
               
               
                             1290                 1295                 1300  
               
               
                   
               
               
                 cac aca cat gac  cag gac aga ctc ggt  gta gtg ctt gac aac  atc      3972  
               
               
                 His Thr His Asp  Gln Asp Arg Leu Gly  Val Val Leu Asp Asn  Ile  
               
               
                             1305                 1310                 1315  
               
               
                   
               
               
                 tat caa ggg tca  acc agg tac gag gca  ggg aga gct cca gcg  tac      4017  
               
               
                 Tyr Gln Gly Ser  Thr Arg Tyr Glu Ala  Gly Arg Ala Pro Ala  Tyr  
               
               
                             1320                 1325                 1330  
               
               
                   
               
               
                 aga gtg atc aga  ggt gac att agc aag  agc gct gac caa gct  atc      4062  
               
               
                 Arg Val Ile Arg  Gly Asp Ile Ser Lys  Ser Ala Asp Gln Ala  Ile  
               
               
                             1335                 1340                 1345  
               
               
                   
               
               
                 gtt aat gct gct  aat agc aaa ggt caa  cca ggt tcc gga gtg  tgc      4107  
               
               
                 Val Asn Ala Ala  Asn Ser Lys Gly Gln  Pro Gly Ser Gly Val  Cys  
               
               
                             1350                 1355                 1360  
               
               
                   
               
               
                 ggt gca ctg tac  cga aaa tgg ccg gct  gct ttt gat aga cag  cca      4152  
               
               
                 Gly Ala Leu Tyr  Arg Lys Trp Pro Ala  Ala Phe Asp Arg Gln  Pro  
               
               
                             1365                 1370                 1375  
               
               
                   
               
               
                 ata gct gtc ggg  acg gct aga ctt gtg  aag cac gaa ccg ctc  atc      4197  
               
               
                 Ile Ala Val Gly  Thr Ala Arg Leu Val  Lys His Glu Pro Leu  Ile  
               
               
                             1380                 1385                 1390  
               
               
                   
               
               
                 ata cat gct gta  gga ccc aat ttt tct  aag atg ccg gaa ccg  gag      4242  
               
               
                 Ile His Ala Val  Gly Pro Asn Phe Ser  Lys Met Pro Glu Pro  Glu  
               
               
                             1395                 1400                 1405  
               
               
                   
               
               
                 ggc gac ctt aag  ctc gca gct gcc tac  atg agc ata gcg tcc  atc      4287  
               
               
                 Gly Asp Leu Lys  Leu Ala Ala Ala Tyr  Met Ser Ile Ala Ser  Ile  
               
               
                             1410                 1415                 1420  
               
               
                   
               
               
                 gtc aac gct gag  cgg att aca aaa ata  tca gta ccg cta ctg  tca      4332  
               
               
                 Val Asn Ala Glu  Arg Ile Thr Lys Ile  Ser Val Pro Leu Leu  Ser  
               
               
                             1425                 1430                 1435  
               
               
                   
               
               
                 acc ggc atc tat  tct ggt ggc aaa gat  cga gtg atg caa tca  ttg      4377  
               
               
                 Thr Gly Ile Tyr  Ser Gly Gly Lys Asp  Arg Val Met Gln Ser  Leu  
               
               
                             1440                 1445                 1450  
               
               
                   
               
               
                 cat cac ctg ttc  act gct ttc gac act  acg gat gcc gat gtc  acc      4422  
               
               
                 His His Leu Phe  Thr Ala Phe Asp Thr  Thr Asp Ala Asp Val  Thr  
               
               
                             1455                 1460                 1465  
               
               
                   
               
               
                 ata tat tgc ttg  gat aaa caa tgg gag  acc agg ata atc gag  gcc      4467  
               
               
                 Ile Tyr Cys Leu  Asp Lys Gln Trp Glu  Thr Arg Ile Ile Glu  Ala  
               
               
                             1470                 1475                 1480  
               
               
                   
               
               
                 att cac cgc aaa  gaa agc gtc gaa att  ctg gat gat gac aag  cca      4512  
               
               
                 Ile His Arg Lys  Glu Ser Val Glu Ile  Leu Asp Asp Asp Lys  Pro  
               
               
                             1485                 1490                 1495  
               
               
                   
               
               
                 gta gac att gac  ttg gtc agg gtc cac  cca aac agc tct ttg  gca      4557  
               
               
                 Val Asp Ile Asp  Leu Val Arg Val His  Pro Asn Ser Ser Leu  Ala  
               
               
                             1500                 1505                 1510  
               
               
                   
               
               
                 ggc aga cca ggt  tac tcc gtc aat gag  ggc aag ttg tat tca  tac      4602  
               
               
                 Gly Arg Pro Gly  Tyr Ser Val Asn Glu  Gly Lys Leu Tyr Ser  Tyr  
               
               
                             1515                 1520                 1525  
               
               
                   
               
               
                 ctg gaa ggt aca  cga ttc cat cag acc  gcc aag gac att gcc  gaa      4647  
               
               
                 Leu Glu Gly Thr  Arg Phe His Gln Thr  Ala Lys Asp Ile Ala  Glu  
               
               
                             1530                 1535                 1540  
               
               
                   
               
               
                 atc cat gca atg  tgg ccc aac aaa tct  gag gct aat gag cag  att      4692  
               
               
                 Ile His Ala Met  Trp Pro Asn Lys Ser  Glu Ala Asn Glu Gln  Ile  
               
               
                             1545                 1550                 1555  
               
               
                   
               
               
                 tgc ttg tac atc  ctg ggg gag agt atg  tcc agc atc cgc tcc  aaa      4737  
               
               
                 Cys Leu Tyr Ile  Leu Gly Glu Ser Met  Ser Ser Ile Arg Ser  Lys  
               
               
                             1560                 1565                 1570  
               
               
                   
               
               
                 tgc cca gta gag  gag tca gag gcg tct  gct cca cct cac aca  ctt      4782  
               
               
                 Cys Pro Val Glu  Glu Ser Glu Ala Ser  Ala Pro Pro His Thr  Leu  
               
               
                             1575                 1580                 1585  
               
               
                   
               
               
                 cca tgc ctg tgt  aat tac gct atg acg  gct gag cgc gta tac  agg      4827  
               
               
                 Pro Cys Leu Cys  Asn Tyr Ala Met Thr  Ala Glu Arg Val Tyr  Arg  
               
               
                             1590                 1595                 1600  
               
               
                   
               
               
                 ttg cgc tct gcg  aag aaa gaa cag ttc  gcc gta tgc tca tca  ttc      4872  
               
               
                 Leu Arg Ser Ala  Lys Lys Glu Gln Phe  Ala Val Cys Ser Ser  Phe  
               
               
                             1605                 1610                 1615  
               
               
                   
               
               
                 ctg ttg ccg aag  tac agg atc aca ggc  gtg cag aag cta cag  tgc      4917  
               
               
                 Leu Leu Pro Lys  Tyr Arg Ile Thr Gly  Val Gln Lys Leu Gln  Cys  
               
               
                             1620                 1625                 1630  
               
               
                   
               
               
                 agc aaa cca gtc  ctg ttt tca ggc gtc  gta cca ccg gct gta  cac      4962  
               
               
                 Ser Lys Pro Val  Leu Phe Ser Gly Val  Val Pro Pro Ala Val  His  
               
               
                             1635                 1640                 1645  
               
               
                   
               
               
                 ccc agg aag tac  gcg gaa ata att cta  gaa acg cca cca ccg  cca      5007  
               
               
                 Pro Arg Lys Tyr  Ala Glu Ile Ile Leu  Glu Thr Pro Pro Pro  Pro  
               
               
                             1650                 1655                 1660  
               
               
                   
               
               
                 gca acg aca acc  gta ata tgt gaa cca  act gtg cca gaa cgt  ata      5052  
               
               
                 Ala Thr Thr Thr  Val Ile Cys Glu Pro  Thr Val Pro Glu Arg  Ile  
               
               
                             1665                 1670                 1675  
               
               
                   
               
               
                 ccc agt ccg gtg  att tct aga gca cca  agt gcg gaa tca ctg  cta      5097  
               
               
                 Pro Ser Pro Val  Ile Ser Arg Ala Pro  Ser Ala Glu Ser Leu  Leu  
               
               
                             1680                 1685                 1690  
               
               
                   
               
               
                 tcg ctt ggc ggc  gtc tcg ttc tct agc  tct gcc aca cgc tcg  tca      5142  
               
               
                 Ser Leu Gly Gly  Val Ser Phe Ser Ser  Ser Ala Thr Arg Ser  Ser  
               
               
                             1695                 1700                 1705  
               
               
                   
               
               
                 acc gcc tgg agc  gac tat gac agg cgg  ttt gtg gtt aca gct  gat      5187  
               
               
                 Thr Ala Trp Ser  Asp Tyr Asp Arg Arg  Phe Val Val Thr Ala  Asp  
               
               
                             1710                 1715                 1720  
               
               
                   
               
               
                 gtg cat caa gcg  aac acg tct acg tgg  agc atc cct agt gct  cct      5232  
               
               
                 Val His Gln Ala  Asn Thr Ser Thr Trp  Ser Ile Pro Ser Ala  Pro  
               
               
                             1725                 1730                 1735  
               
               
                   
               
               
                 ggc ttg gac gtc  cag ctg cct tct gac  gtc act gat tcc cac  tgg      5277  
               
               
                 Gly Leu Asp Val  Gln Leu Pro Ser Asp  Val Thr Asp Ser His  Trp  
               
               
                             1740                 1745                 1750  
               
               
                   
               
               
                 agt att cca agt  gca tca ggc ttt gaa  gtg aga aca cca tct  gta      5322  
               
               
                 Ser Ile Pro Ser  Ala Ser Gly Phe Glu  Val Arg Thr Pro Ser  Val  
               
               
                             1755                 1760                 1765  
               
               
                   
               
               
                 cag gac cta act  gcg gag tgt gcg aag  cct cgt gga ctg gcc  gaa      5367  
               
               
                 Gln Asp Leu Thr  Ala Glu Cys Ala Lys  Pro Arg Gly Leu Ala  Glu  
               
               
                             1770                 1775                 1780  
               
               
                   
               
               
                 ata atg caa gac  ttc aat act gct cct  ttc cag ttt ctt tcg  gac      5412  
               
               
                 Ile Met Gln Asp  Phe Asn Thr Ala Pro  Phe Gln Phe Leu Ser  Asp  
               
               
                             1785                 1790                 1795  
               
               
                   
               
               
                 tac aga cca gta  ccg gca cca cgg aga  cgc ccc atc cca tca  cct      5457  
               
               
                 Tyr Arg Pro Val  Pro Ala Pro Arg Arg  Arg Pro Ile Pro Ser  Pro  
               
               
                             1800                 1805                 1810  
               
               
                   
               
               
                 aga tcg acg gct  tcc gca cct cca gtt  cca aag cca cgc agg  act      5502  
               
               
                 Arg Ser Thr Ala  Ser Ala Pro Pro Val  Pro Lys Pro Arg Arg  Thr  
               
               
                             1815                 1820                 1825  
               
               
                   
               
               
                 aag tac caa caa  cca cca gga gtc gct  aga gcg atc tca gaa  gcg      5547  
               
               
                 Lys Tyr Gln Gln  Pro Pro Gly Val Ala  Arg Ala Ile Ser Glu  Ala  
               
               
                             1830                 1835                 1840  
               
               
                   
               
               
                 gag ttg gac gag  tac atc cgt caa cac  tcc aac tga cgg tat gaa       5592  
               
               
                 Glu Leu Asp Glu  Tyr Ile Arg Gln His  Ser Asn     Arg Tyr Glu  
               
               
                             1845                 1850                     1855  
               
               
                   
               
               
                 gcg gga gcg tat att  ttc tca tcg gaa aca  ggc caa ggt cac ctt       5637  
               
               
                 Ala Gly Ala Tyr Ile  Phe Ser Ser Glu Thr  Gly Gln Gly His Leu  
               
               
                                 1860                 1865                 1870  
               
               
                   
               
               
                 caa cag aaa tca gta  cgt caa tgt aaa cta  caa gaa cct ata ttg       5682  
               
               
                 Gln Gln Lys Ser Val  Arg Gln Cys Lys Leu  Gln Glu Pro Ile Leu  
               
               
                                 1875                 1880                 1885  
               
               
                   
               
               
                 gat cgg gcc gtc cat  gag aag tat tac gcc  ccg cgc ctc gat ctc       5727  
               
               
                 Asp Arg Ala Val His  Glu Lys Tyr Tyr Ala  Pro Arg Leu Asp Leu  
               
               
                                 1890                 1895                 1900  
               
               
                   
               
               
                 gaa aga gag aaa atg  tta cag aag aaa ctg  caa tta tgc gcc tct       5772  
               
               
                 Glu Arg Glu Lys Met  Leu Gln Lys Lys Leu  Gln Leu Cys Ala Ser  
               
               
                                 1905                 1910                 1915  
               
               
                   
               
               
                 gaa gga aat aga agc  agg tat caa tca cga  aaa gta gaa aat atg       5817  
               
               
                 Glu Gly Asn Arg Ser  Arg Tyr Gln Ser Arg  Lys Val Glu Asn Met  
               
               
                                 1920                 1925                 1930  
               
               
                   
               
               
                 aaa gca att aca gcg  gag cga ctc att tct  gga ttg ggc aca tat       5862  
               
               
                 Lys Ala Ile Thr Ala  Glu Arg Leu Ile Ser  Gly Leu Gly Thr Tyr  
               
               
                                 1935                 1940                 1945  
               
               
                   
               
               
                 cta tca tca gaa gtg  aat cct gtc gag tgt  tac aga gtc aat tat       5907  
               
               
                 Leu Ser Ser Glu Val  Asn Pro Val Glu Cys  Tyr Arg Val Asn Tyr  
               
               
                                 1950                 1955                 1960  
               
               
                   
               
               
                 cct gta cca atc tac  tcg tca acg gta att  aac agg ttt aca tct       5952  
               
               
                 Pro Val Pro Ile Tyr  Ser Ser Thr Val Ile  Asn Arg Phe Thr Ser  
               
               
                                 1965                 1970                 1975  
               
               
                   
               
               
                 gca gag gtc gcg gtt  aaa acg tgc aac tta  gtt atc caa gag aat       5997  
               
               
                 Ala Glu Val Ala Val  Lys Thr Cys Asn Leu  Val Ile Gln Glu Asn  
               
               
                                 1980                 1985                 1990  
               
               
                   
               
               
                 tac cct aca gta gcc  agt tat tgt ata aca  gat gaa tac gat gcg       6042  
               
               
                 Tyr Pro Thr Val Ala  Ser Tyr Cys Ile Thr  Asp Glu Tyr Asp Ala  
               
               
                                 1995                 2000                 2005  
               
               
                   
               
               
                 tat ctt gac atg gtg  gac ggc gca tcg tgc  tgt cta gat aca gcc       6087  
               
               
                 Tyr Leu Asp Met Val  Asp Gly Ala Ser Cys  Cys Leu Asp Thr Ala  
               
               
                                 2010                 2015                 2020  
               
               
                   
               
               
                 act ttt tgt ccg gct  aaa ctg aga agc tac  cca aag aag cat agc       6132  
               
               
                 Thr Phe Cys Pro Ala  Lys Leu Arg Ser Tyr  Pro Lys Lys His Ser  
               
               
                                 2025                 2030                 2035  
               
               
                   
               
               
                 tat ttg cag cca gag  ata aga tca gcc gtc  cca tcg cct ata cag       6177  
               
               
                 Tyr Leu Gln Pro Glu  Ile Arg Ser Ala Val  Pro Ser Pro Ile Gln  
               
               
                                 2040                 2045                 2050  
               
               
                   
               
               
                 aat aca tta caa aat  gta ttg gct gca gct  act aaa agg aat tgc       6222  
               
               
                 Asn Thr Leu Gln Asn  Val Leu Ala Ala Ala  Thr Lys Arg Asn Cys  
               
               
                                 2055                 2060                 2065  
               
               
                   
               
               
                 aac gtt acc caa atg  cga gaa tta cct gtc  tta gat tcg gcg gca       6267  
               
               
                 Asn Val Thr Gln Met  Arg Glu Leu Pro Val  Leu Asp Ser Ala Ala  
               
               
                                 2070                 2075                 2080  
               
               
                   
               
               
                 ttt aat gtt gat tgt  ttc aag aaa tac gca  tgc aat gat gag tac       6312  
               
               
                 Phe Asn Val Asp Cys  Phe Lys Lys Tyr Ala  Cys Asn Asp Glu Tyr  
               
               
                                 2085                 2090                 2095  
               
               
                   
               
               
                 tgg gat acc ttt cgc  gat aac cct att cgg  cta act aca gag aac       6357  
               
               
                 Trp Asp Thr Phe Arg  Asp Asn Pro Ile Arg  Leu Thr Thr Glu Asn  
               
               
                                 2100                 2105                 2110  
               
               
                   
               
               
                 gtt acg caa tat gtg  aca aag ctg aaa ggg  ccg aaa gca gca gca       6402  
               
               
                 Val Thr Gln Tyr Val  Thr Lys Leu Lys Gly  Pro Lys Ala Ala Ala  
               
               
                                 2115                 2120                 2125  
               
               
                   
               
               
                 ttg ttt gcg aat act  cat aat cta aaa ccg  ttg cag gag ata cca       6447  
               
               
                 Leu Phe Ala Asn Thr  His Asn Leu Lys Pro  Leu Gln Glu Ile Pro  
               
               
                                 2130                 2135                 2140  
               
               
                   
               
               
                 atg gat caa ttc gtc  atg gat cta aag aga  gat gtc aaa gtt act       6492  
               
               
                 Met Asp Gln Phe Val  Met Asp Leu Lys Arg  Asp Val Lys Val Thr  
               
               
                                 2145                 2150                 2155  
               
               
                   
               
               
                 ccc ggc acg aaa cat  aca gag gag cgg cct  aag gtg cag gtt att       6537  
               
               
                 Pro Gly Thr Lys His  Thr Glu Glu Arg Pro  Lys Val Gln Val Ile  
               
               
                                 2160                 2165                 2170  
               
               
                   
               
               
                 cag gct gca gat ccc  ctt gct acc gct tac  ctt tgc ggg atc cat       6582  
               
               
                 Gln Ala Ala Asp Pro  Leu Ala Thr Ala Tyr  Leu Cys Gly Ile His  
               
               
                                 2175                 2180                 2185  
               
               
                   
               
               
                 cgg gaa tta gtc cgt  aga ctg aat gcg gtg  ctt ctg cca aat atc       6627  
               
               
                 Arg Glu Leu Val Arg  Arg Leu Asn Ala Val  Leu Leu Pro Asn Ile  
               
               
                                 2190                 2195                 2200  
               
               
                   
               
               
                 cat act ctc ttc gac  atg tca gcg gaa gat  ttt gat gcg att att       6672  
               
               
                 His Thr Leu Phe Asp  Met Ser Ala Glu Asp  Phe Asp Ala Ile Ile  
               
               
                                 2205                 2210                 2215  
               
               
                   
               
               
                 gct gaa cat ttc cac  cac ggc gac cca gta  ttg gaa acg gac atc       6717  
               
               
                 Ala Glu His Phe His  His Gly Asp Pro Val  Leu Glu Thr Asp Ile  
               
               
                                 2220                 2225                 2230  
               
               
                   
               
               
                 gcg tcg ttt gat aaa  agc gaa gac gac gct  atc gcc att tcg gcg       6762  
               
               
                 Ala Ser Phe Asp Lys  Ser Glu Asp Asp Ala  Ile Ala Ile Ser Ala  
               
               
                                 2235                 2240                 2245  
               
               
                   
               
               
                 ttg atg atc ctt gag  gac tta ggt gtc gac  caa ccg ctc tta gat       6807  
               
               
                 Leu Met Ile Leu Glu  Asp Leu Gly Val Asp  Gln Pro Leu Leu Asp  
               
               
                                 2250                 2255                 2260  
               
               
                   
               
               
                 ttg ata gag gcg gcg  ttc ggc aat atc aca  tct gtg cac cta cct       6852  
               
               
                 Leu Ile Glu Ala Ala  Phe Gly Asn Ile Thr  Ser Val His Leu Pro  
               
               
                                 2265                 2270                 2275  
               
               
                   
               
               
                 aca gga acg agg ttt  aaa ttt ggt gcc atg  atg aaa tcc ggt atg       6897  
               
               
                 Thr Gly Thr Arg Phe  Lys Phe Gly Ala Met  Met Lys Ser Gly Met  
               
               
                                 2280                 2285                 2290  
               
               
                   
               
               
                 ttc tta acg ctg ttt  gtc aac aca cta gtc  aat atc atg att gct       6942  
               
               
                 Phe Leu Thr Leu Phe  Val Asn Thr Leu Val  Asn Ile Met Ile Ala  
               
               
                                 2295                 2300                 2305  
               
               
                   
               
               
                 agc aga gta cta cgt  gaa cgg tta acc acg  tca gcg tgc gcg gcc       6987  
               
               
                 Ser Arg Val Leu Arg  Glu Arg Leu Thr Thr  Ser Ala Cys Ala Ala  
               
               
                                 2310                 2315                 2320  
               
               
                   
               
               
                 tct atc ggc gac gat  aac ata gtg cat ggt  gtc gtc tcc gac acc       7032  
               
               
                 Ser Ile Gly Asp Asp  Asn Ile Val His Gly  Val Val Ser Asp Thr  
               
               
                                 2325                 2330                 2335  
               
               
                   
               
               
                 ttg atg gcg gag aga  tgc gcc act tgg ctg  aac atg gaa gta aaa       7077  
               
               
                 Leu Met Ala Glu Arg  Cys Ala Thr Trp Leu  Asn Met Glu Val Lys  
               
               
                                 2340                 2345                 2350  
               
               
                   
               
               
                 att att gat gca gtt  att ggt atc aaa gca  ccc tac ttc tgt ggg       7122  
               
               
                 Ile Ile Asp Ala Val  Ile Gly Ile Lys Ala  Pro Tyr Phe Cys Gly  
               
               
                                 2355                 2360                 2365  
               
               
                   
               
               
                 gga ttt atc ctg gtg  gac cag ata aca ggc  aca gcc tgc aga gtc       7167  
               
               
                 Gly Phe Ile Leu Val  Asp Gln Ile Thr Gly  Thr Ala Cys Arg Val  
               
               
                                 2370                 2375                 2380  
               
               
                   
               
               
                 gca gac cct cta aaa  agg ctt ttt aag ctt  gga aaa cca ttg cca       7212  
               
               
                 Ala Asp Pro Leu Lys  Arg Leu Phe Lys Leu  Gly Lys Pro Leu Pro  
               
               
                                 2385                 2390                 2395  
               
               
                   
               
               
                 gtc gat gat acc caa  gac tgc gac cgc cgc  cgg gca ctg cat gat       7257  
               
               
                 Val Asp Asp Thr Gln  Asp Cys Asp Arg Arg  Arg Ala Leu His Asp  
               
               
                                 2400                 2405                 2410  
               
               
                   
               
               
                 gaa gca atg cga tgg  aac aga att gga att  acg gac gag tta gtg       7302  
               
               
                 Glu Ala Met Arg Trp  Asn Arg Ile Gly Ile  Thr Asp Glu Leu Val  
               
               
                                 2415                 2420                 2425  
               
               
                   
               
               
                 aag gcc gta gaa tcc  aga tac gag atc ata  ctg gca ggc ctg atc       7347  
               
               
                 Lys Ala Val Glu Ser  Arg Tyr Glu Ile Ile  Leu Ala Gly Leu Ile  
               
               
                                 2430                 2435                 2440  
               
               
                   
               
               
                 atc acg tct ctg tcc  acg tta gcc gaa agc  gtt aag aac ttc aag       7392  
               
               
                 Ile Thr Ser Leu Ser  Thr Leu Ala Glu Ser  Val Lys Asn Phe Lys  
               
               
                                 2445                 2450                 2455  
               
               
                   
               
               
                 agc ata aga ggg agc  cca atc acc ctc tac  ggc tga cctaaatagg        7438  
               
               
                 Ser Ile Arg Gly Ser  Pro Ile Thr Leu Tyr  Gly  
               
               
                                 2460                 2465  
               
               
                   
               
               
                 tgacgtagta gacacgcacc tacccaccgg caga atg ttt cca tac  cct cag      7490  
               
               
                                                       Met Phe Pro Tyr  Pro Gln  
               
               
                                                                   2470  
               
               
                   
               
               
                 ctg aac ttt  cca cca gtt tac cct  aca aat ccg atg gct  tac cga      7535  
               
               
                 Leu Asn Phe  Pro Pro Val Tyr Pro  Thr Asn Pro Met Ala  Tyr Arg  
               
               
                         2475                 2480                 2485  
               
               
                   
               
               
                 gat cca aac  cct cct agg cgc cgc  tgg agg ccg ttt cgg  ccc ccg      7580  
               
               
                 Asp Pro Asn  Pro Pro Arg Arg Arg  Trp Arg Pro Phe Arg  Pro Pro  
               
               
                         2490                 2495                 2500  
               
               
                   
               
               
                 ctg gct gct  caa atc gaa gat ctt  agg agg tcg ata gtc  aac ttg      7625  
               
               
                 Leu Ala Ala  Gln Ile Glu Asp Leu  Arg Arg Ser Ile Val  Asn Leu  
               
               
                         2505                 2510                 2515  
               
               
                   
               
               
                 act ttc aaa  caa cga tca cct aat  ccg ccg cca ggt cca  ccg cca      7670  
               
               
                 Thr Phe Lys  Gln Arg Ser Pro Asn  Pro Pro Pro Gly Pro  Pro Pro  
               
               
                         2520                 2525                 2530  
               
               
                   
               
               
                 aag aag aag  aag agt gct cct aag  cca aaa cct act cag  cct aaa      7715  
               
               
                 Lys Lys Lys  Lys Ser Ala Pro Lys  Pro Lys Pro Thr Gln  Pro Lys  
               
               
                         2535                 2540                 2545  
               
               
                   
               
               
                 aag aag aag  cag caa gcc aag agg  acg aaa cgc aag cct  aaa cca      7760  
               
               
                 Lys Lys Lys  Gln Gln Ala Lys Arg  Thr Lys Arg Lys Pro  Lys Pro  
               
               
                         2550                 2555                 2560  
               
               
                   
               
               
                 ggg aaa cga  caa cgt atg tgt atg  aag ttg gag tcg gac  aag aca      7805  
               
               
                 Gly Lys Arg  Gln Arg Met Cys Met  Lys Leu Glu Ser Asp  Lys Thr  
               
               
                         2565                 2570                 2575  
               
               
                   
               
               
                 ttt ccg atc  atg ctg aac ggc caa  gtg aat gga tat gcc  tgc gtt      7850  
               
               
                 Phe Pro Ile  Met Leu Asn Gly Gln  Val Asn Gly Tyr Ala  Cys Val  
               
               
                         2580                 2585                 2590  
               
               
                   
               
               
                 gtc gga gga  agg ctg atg aaa cca  ctc cac gtt gaa gga  aaa att      7895  
               
               
                 Val Gly Gly  Arg Leu Met Lys Pro  Leu His Val Glu Gly  Lys Ile  
               
               
                         2595                 2600                 2605  
               
               
                   
               
               
                 gat aat gag  caa tta gcg gcc gtg  aaa ttg aag aag gct  agc atg      7940  
               
               
                 Asp Asn Glu  Gln Leu Ala Ala Val  Lys Leu Lys Lys Ala  Ser Met  
               
               
                         2610                 2615                 2620  
               
               
                   
               
               
                 tac gac ttg  gag tac ggc gac gtt  ccc cag aac atg aaa  tca gac      7985  
               
               
                 Tyr Asp Leu  Glu Tyr Gly Asp Val  Pro Gln Asn Met Lys  Ser Asp  
               
               
                         2625                 2630                 2635  
               
               
                   
               
               
                 acg ctg cag  tac acc agc gac aaa  cca ccg ggc ttc tac  aac tgg      8030  
               
               
                 Thr Leu Gln  Tyr Thr Ser Asp Lys  Pro Pro Gly Phe Tyr  Asn Trp  
               
               
                         2640                 2645                 2650  
               
               
                   
               
               
                 cac cac ggc  gca gtc cag tat gag  aat ggg aga ttt acc  gta ccg      8075  
               
               
                 His His Gly  Ala Val Gln Tyr Glu  Asn Gly Arg Phe Thr  Val Pro  
               
               
                         2655                 2660                 2665  
               
               
                   
               
               
                 aga gga gtg  ggc ggg aaa ggc gac  agc gga aga ccg atc  ctg gac      8120  
               
               
                 Arg Gly Val  Gly Gly Lys Gly Asp  Ser Gly Arg Pro Ile  Leu Asp  
               
               
                         2670                 2675                 2680  
               
               
                   
               
               
                 aac aga ggc  aga gtt gtg gct att  gtt cta gga ggt gca  aat gag      8165  
               
               
                 Asn Arg Gly  Arg Val Val Ala Ile  Val Leu Gly Gly Ala  Asn Glu  
               
               
                         2685                 2690                 2695  
               
               
                   
               
               
                 ggc acg cgt  acg gcg ctt tca gtg  gtc act tgg aac cag  aaa ggg      8210  
               
               
                 Gly Thr Arg  Thr Ala Leu Ser Val  Val Thr Trp Asn Gln  Lys Gly  
               
               
                         2700                 2705                 2710  
               
               
                   
               
               
                 gtg acc att  agg gat acc ccc gaa  ggt tct gaa ccg tgg  tca cta      8255  
               
               
                 Val Thr Ile  Arg Asp Thr Pro Glu  Gly Ser Glu Pro Trp  Ser Leu  
               
               
                         2715                 2720                 2725  
               
               
                   
               
               
                 gtt aca gcg  cta tgc gtg ctt tcg  aat gtc acg ttc cca  tgc gac      8300  
               
               
                 Val Thr Ala  Leu Cys Val Leu Ser  Asn Val Thr Phe Pro  Cys Asp  
               
               
                         2730                 2735                 2740  
               
               
                   
               
               
                 aaa cca ccc  gtg tgc tat tca ctg  acg cca gaa cga aca  ctc gac      8345  
               
               
                 Lys Pro Pro  Val Cys Tyr Ser Leu  Thr Pro Glu Arg Thr  Leu Asp  
               
               
                         2745                 2750                 2755  
               
               
                   
               
               
                 gtg ctc gaa  gag aac gtc gac aat  cca aat tac gac acg  ctg ctg      8390  
               
               
                 Val Leu Glu  Glu Asn Val Asp Asn  Pro Asn Tyr Asp Thr  Leu Leu  
               
               
                         2760                 2765                 2770  
               
               
                   
               
               
                 gag aac gtc  ttg aaa tgt cca tca  cgc cgg ccc aaa cga  agc att      8435  
               
               
                 Glu Asn Val  Leu Lys Cys Pro Ser  Arg Arg Pro Lys Arg  Ser Ile  
               
               
                         2775                 2780                 2785  
               
               
                   
               
               
                 acc gat gac  ttc aca ctg acc agt  ccc tac ctg ggg ttc  tgc ccg      8480  
               
               
                 Thr Asp Asp  Phe Thr Leu Thr Ser  Pro Tyr Leu Gly Phe  Cys Pro  
               
               
                         2790                 2795                 2800  
               
               
                   
               
               
                 tat tgc aga  cac tca acg ccg tgt  ttc agc cca ata aaa  att gag      8525  
               
               
                 Tyr Cys Arg  His Ser Thr Pro Cys  Phe Ser Pro Ile Lys  Ile Glu  
               
               
                         2805                 2810                 2815  
               
               
                   
               
               
                 aac gtg tgg  gac gaa tct gat gat  gga tcg att aga atc  cag gtc      8570  
               
               
                 Asn Val Trp  Asp Glu Ser Asp Asp  Gly Ser Ile Arg Ile  Gln Val  
               
               
                         2820                 2825                 2830  
               
               
                   
               
               
                 tcg gca caa  ttc ggc tac aat cag  gca ggc act gcg gat  gtc acc      8615  
               
               
                 Ser Ala Gln  Phe Gly Tyr Asn Gln  Ala Gly Thr Ala Asp  Val Thr  
               
               
                         2835                 2840                 2845  
               
               
                   
               
               
                 aaa ttc cgt  tac atg tct ttc gac  cac gac cat gac atc  aag gaa      8660  
               
               
                 Lys Phe Arg  Tyr Met Ser Phe Asp  His Asp His Asp Ile  Lys Glu  
               
               
                         2850                 2855                 2860  
               
               
                   
               
               
                 gac agt atg  gag aaa ata gct atc  agc aca tct gga ccc  tgc cgt      8705  
               
               
                 Asp Ser Met  Glu Lys Ile Ala Ile  Ser Thr Ser Gly Pro  Cys Arg  
               
               
                         2865                 2870                 2875  
               
               
                   
               
               
                 cgt ctt ggc  cac aaa ggg tac ttc  ctg tta gct caa tgt  cct cca      8750  
               
               
                 Arg Leu Gly  His Lys Gly Tyr Phe  Leu Leu Ala Gln Cys  Pro Pro  
               
               
                         2880                 2885                 2890  
               
               
                   
               
               
                 ggt gac agt  gta acc gtc agt atc  acg agc gga gca tct  gag aat      8795  
               
               
                 Gly Asp Ser  Val Thr Val Ser Ile  Thr Ser Gly Ala Ser  Glu Asn  
               
               
                         2895                 2900                 2905  
               
               
                   
               
               
                 tca tgc acc  gtg gag aaa aag atc  agg agg aag ttt gtc  ggt aga      8840  
               
               
                 Ser Cys Thr  Val Glu Lys Lys Ile  Arg Arg Lys Phe Val  Gly Arg  
               
               
                         2910                 2915                 2920  
               
               
                   
               
               
                 gag gag tac  ttg ttc cca ccc gtc  cat gga aag ctg gta  aag tgc      8885  
               
               
                 Glu Glu Tyr  Leu Phe Pro Pro Val  His Gly Lys Leu Val  Lys Cys  
               
               
                         2925                 2930                 2935  
               
               
                   
               
               
                 cac gtt tac  gat cac ttg aag gag  acg tct gcc ggg tac  ata acc      8930  
               
               
                 His Val Tyr  Asp His Leu Lys Glu  Thr Ser Ala Gly Tyr  Ile Thr  
               
               
                         2940                 2945                 2950  
               
               
                   
               
               
                 atg cac agg  cca ggc cca cac gcg  tat aag tcc tat ctg  gag gaa      8975  
               
               
                 Met His Arg  Pro Gly Pro His Ala  Tyr Lys Ser Tyr Leu  Glu Glu  
               
               
                         2955                 2960                 2965  
               
               
                   
               
               
                 gcg tca ggc  gaa gtg tac att aaa  cca cct tct ggc aag  aac gtc      9020  
               
               
                 Ala Ser Gly  Glu Val Tyr Ile Lys  Pro Pro Ser Gly Lys  Asn Val  
               
               
                         2970                 2975                 2980  
               
               
                   
               
               
                 acc tac gaa  tgt aag tgt ggc gac  tac agc aca ggt atc  gtg agc      9065  
               
               
                 Thr Tyr Glu  Cys Lys Cys Gly Asp  Tyr Ser Thr Gly Ile  Val Ser  
               
               
                         2985                 2990                 2995  
               
               
                   
               
               
                 acg cga acg  aag atg aac ggc tgc  act aaa gca aaa cag  tgc att      9110  
               
               
                 Thr Arg Thr  Lys Met Asn Gly Cys  Thr Lys Ala Lys Gln  Cys Ile  
               
               
                         3000                 3005                 3010  
               
               
                   
               
               
                 gcc tac aag  agc gac caa acg aaa  tgg gtc ttc aac tcg  ccg gat      9155  
               
               
                 Ala Tyr Lys  Ser Asp Gln Thr Lys  Trp Val Phe Asn Ser  Pro Asp  
               
               
                         3015                 3020                 3025  
               
               
                   
               
               
                 ctt att agg  cac aca gac cac tca  gtg caa ggt aaa ttg  cac att      9200  
               
               
                 Leu Ile Arg  His Thr Asp His Ser  Val Gln Gly Lys Leu  His Ile  
               
               
                         3030                 3035                 3040  
               
               
                   
               
               
                 cca ttc cgc  ttg aca ccg aca gtc  tgc ccg gtt ccg tta  gct cac      9245  
               
               
                 Pro Phe Arg  Leu Thr Pro Thr Val  Cys Pro Val Pro Leu  Ala His  
               
               
                         3045                 3050                 3055  
               
               
                   
               
               
                 acg cct aca  gtc acg aag tgg ttc  aaa ggc atc acc ctc  cac ctg      9290  
               
               
                 Thr Pro Thr  Val Thr Lys Trp Phe  Lys Gly Ile Thr Leu  His Leu  
               
               
                         3060                 3065                 3070  
               
               
                   
               
               
                 act gca atg  cga cca aca ttg ctg  aca acg aga aaa ttg  ggg ctg      9335  
               
               
                 Thr Ala Met  Arg Pro Thr Leu Leu  Thr Thr Arg Lys Leu  Gly Leu  
               
               
                         3075                 3080                 3085  
               
               
                   
               
               
                 cga gca gac  gca aca gca gaa tgg  att aca ggg tct aca  tcc agg      9380  
               
               
                 Arg Ala Asp  Ala Thr Ala Glu Trp  Ile Thr Gly Ser Thr  Ser Arg  
               
               
                         3090                 3095                 3100  
               
               
                   
               
               
                 aat ttt tct  gtg ggg cga gaa ggg  ctg gag tac gta tgg  ggt aac      9425  
               
               
                 Asn Phe Ser  Val Gly Arg Glu Gly  Leu Glu Tyr Val Trp  Gly Asn  
               
               
                         3105                 3110                 3115  
               
               
                   
               
               
                 cat gaa cca  gtc aga gtc tgg gcc  cag gag tcg gca cca  ggc gac      9470  
               
               
                 His Glu Pro  Val Arg Val Trp Ala  Gln Glu Ser Ala Pro  Gly Asp  
               
               
                         3120                 3125                 3130  
               
               
                   
               
               
                 cca cat gga  tgg ccg cat gag atc  atc atc cac tat tat  cat cgg      9515  
               
               
                 Pro His Gly  Trp Pro His Glu Ile  Ile Ile His Tyr Tyr  His Arg  
               
               
                         3135                 3140                 3145  
               
               
                   
               
               
                 cat cca gtc  tac act gtc att gtg  ctg tgt ggt gtc gct  ctt gct      9560  
               
               
                 His Pro Val  Tyr Thr Val Ile Val  Leu Cys Gly Val Ala  Leu Ala  
               
               
                         3150                 3155                 3160  
               
               
                   
               
               
                 atc ctg gta  ggc act gca tca tca  gca gct tgc atc gcc  aaa gca      9605  
               
               
                 Ile Leu Val  Gly Thr Ala Ser Ser  Ala Ala Cys Ile Ala  Lys Ala  
               
               
                         3165                 3170                 3175  
               
               
                   
               
               
                 aga aga gac  tgc ctg acg cca tac  gcg ctt gca ccg aac  gca acg      9650  
               
               
                 Arg Arg Asp  Cys Leu Thr Pro Tyr  Ala Leu Ala Pro Asn  Ala Thr  
               
               
                         3180                 3185                 3190  
               
               
                   
               
               
                 gta ccc aca  gca tta gcg gtt ttg  tgc tgc att cgg cca  acc aac      9695  
               
               
                 Val Pro Thr  Ala Leu Ala Val Leu  Cys Cys Ile Arg Pro  Thr Asn  
               
               
                         3195                 3200                 3205  
               
               
                   
               
               
                 gct gaa aca  ttt gga gaa act ttg  aac cat ctg tgg ttt  aac aac      9740  
               
               
                 Ala Glu Thr  Phe Gly Glu Thr Leu  Asn His Leu Trp Phe  Asn Asn  
               
               
                         3210                 3215                 3220  
               
               
                   
               
               
                 caa ccg ttt  ctc tgg gca cag ttg  tgc att cct ctg gca  gcg ctt      9785  
               
               
                 Gln Pro Phe  Leu Trp Ala Gln Leu  Cys Ile Pro Leu Ala  Ala Leu  
               
               
                         3225                 3230                 3235  
               
               
                   
               
               
                 gtt att ctg  ttc cgc tgc ttt tca  tgc tgc atg cct ttt  tta ttg      9830  
               
               
                 Val Ile Leu  Phe Arg Cys Phe Ser  Cys Cys Met Pro Phe  Leu Leu  
               
               
                         3240                 3245                 3250  
               
               
                   
               
               
                 gtt gca ggc  gtc tgc ctg ggg aag  gta gac gcc ttc gaa  cat gcg      9875  
               
               
                 Val Ala Gly  Val Cys Leu Gly Lys  Val Asp Ala Phe Glu  His Ala  
               
               
                         3255                 3260                 3265  
               
               
                   
               
               
                 acc act gtg  cca aat gtt ccg ggg  atc ccg tat aag gcg  ttg gtc      9920  
               
               
                 Thr Thr Val  Pro Asn Val Pro Gly  Ile Pro Tyr Lys Ala  Leu Val  
               
               
                         3270                 3275                 3280  
               
               
                   
               
               
                 gaa cgc gca  ggt tac gcg cca ctt  aac ctg gag atc acg  gtc gtc      9965  
               
               
                 Glu Arg Ala  Gly Tyr Ala Pro Leu  Asn Leu Glu Ile Thr  Val Val  
               
               
                         3285                 3290                 3295  
               
               
                   
               
               
                 tca tcg gaa  tta aca cct tca act  aac aag gag tac gtg  acc tgc     10010  
               
               
                 Ser Ser Glu  Leu Thr Pro Ser Thr  Asn Lys Glu Tyr Val  Thr Cys  
               
               
                         3300                 3305                 3310  
               
               
                   
               
               
                 aaa ttc cac  aca gtc att cct tca  cca caa gtt aaa tgc  tgc ggg     10055  
               
               
                 Lys Phe His  Thr Val Ile Pro Ser  Pro Gln Val Lys Cys  Cys Gly  
               
               
                         3315                 3320                 3325  
               
               
                   
               
               
                 tcc ctc gag  tgc aag gca tcc tca  aag gcg gat tac aca  tgc cgc     10100  
               
               
                 Ser Leu Glu  Cys Lys Ala Ser Ser  Lys Ala Asp Tyr Thr  Cys Arg  
               
               
                         3330                 3335                 3340  
               
               
                   
               
               
                 gtt ttt ggc  ggt gtg tac cct ttc  atg tgg gga ggc gca  caa tgc     10145  
               
               
                 Val Phe Gly  Gly Val Tyr Pro Phe  Met Trp Gly Gly Ala  Gln Cys  
               
               
                         3345                 3350                 3355  
               
               
                   
               
               
                 ttc tgt gac  agt gag aac aca caa  ctg agt gag gcg tac  gtc gag     10190  
               
               
                 Phe Cys Asp  Ser Glu Asn Thr Gln  Leu Ser Glu Ala Tyr  Val Glu  
               
               
                         3360                 3365                 3370  
               
               
                   
               
               
                 ttc gct cca  gac tgc act ata gat  cac gca gtc gca cta  aaa gtt     10235  
               
               
                 Phe Ala Pro  Asp Cys Thr Ile Asp  His Ala Val Ala Leu  Lys Val  
               
               
                         3375                 3380                 3385  
               
               
                   
               
               
                 cac aca gct  gct ctg aaa gtc ggc  ctg cgt ata gta tac  ggc aac     10280  
               
               
                 His Thr Ala  Ala Leu Lys Val Gly  Leu Arg Ile Val Tyr  Gly Asn  
               
               
                         3390                 3395                 3400  
               
               
                   
               
               
                 acc acc gcg  cac ctg gat acg ttt  gtc aat ggc gtc acg  cca ggt     10325  
               
               
                 Thr Thr Ala  His Leu Asp Thr Phe  Val Asn Gly Val Thr  Pro Gly  
               
               
                         3405                 3410                 3415  
               
               
                   
               
               
                 tcc tca cgg  gac ctg aag gtc ata  gca ggg ccg ata tca  gcc gct     10370  
               
               
                 Ser Ser Arg  Asp Leu Lys Val Ile  Ala Gly Pro Ile Ser  Ala Ala  
               
               
                         3420                 3425                 3430  
               
               
                   
               
               
                 ttt tca ccc  ttt gac cat aag gtc  gtc atc aga aag ggg  ctt gtt     10415  
               
               
                 Phe Ser Pro  Phe Asp His Lys Val  Val Ile Arg Lys Gly  Leu Val  
               
               
                         3435                 3440                 3445  
               
               
                   
               
               
                 tac aac tac  gac ttc cct gag tat  gga gct atg aaa cca  gga gcg     10460  
               
               
                 Tyr Asn Tyr  Asp Phe Pro Glu Tyr  Gly Ala Met Lys Pro  Gly Ala  
               
               
                         3450                 3455                 3460  
               
               
                   
               
               
                 ttc ggc gat  att caa gca tcc tcg  ctt gat gct aca gac  ata gta     10505  
               
               
                 Phe Gly Asp  Ile Gln Ala Ser Ser  Leu Asp Ala Thr Asp  Ile Val  
               
               
                         3465                 3470                 3475  
               
               
                   
               
               
                 gcc cgc act  gac ata cgg ctg ctg  aag cct tct gtc aag  aac atc     10550  
               
               
                 Ala Arg Thr  Asp Ile Arg Leu Leu  Lys Pro Ser Val Lys  Asn Ile  
               
               
                         3480                 3485                 3490  
               
               
                   
               
               
                 cac gtc ccc  tac acc caa gca gta  tca ggg tat gaa atg  tgg aag     10595  
               
               
                 His Val Pro  Tyr Thr Gln Ala Val  Ser Gly Tyr Glu Met  Trp Lys  
               
               
                         3495                 3500                 3505  
               
               
                   
               
               
                 aac aac tca  gga cga ccc ctg caa  gaa aca gca cca ttt  gga tgt     10640  
               
               
                 Asn Asn Ser  Gly Arg Pro Leu Gln  Glu Thr Ala Pro Phe  Gly Cys  
               
               
                         3510                 3515                 3520  
               
               
                   
               
               
                 aaa att gaa  gtg gag cct ctg cga  gcg tct aac tgt gct  tac ggg     10685  
               
               
                 Lys Ile Glu  Val Glu Pro Leu Arg  Ala Ser Asn Cys Ala  Tyr Gly  
               
               
                         3525                 3530                 3535  
               
               
                   
               
               
                 cac atc cct  atc tcg att gac atc  cct gat gca gct ttt  gtg aga     10730  
               
               
                 His Ile Pro  Ile Ser Ile Asp Ile  Pro Asp Ala Ala Phe  Val Arg  
               
               
                         3540                 3545                 3550  
               
               
                   
               
               
                 tca tca gaa  tca cca aca att tta  gaa gtt agc tgc aca  gta gca     10775  
               
               
                 Ser Ser Glu  Ser Pro Thr Ile Leu  Glu Val Ser Cys Thr  Val Ala  
               
               
                         3555                 3560                 3565  
               
               
                   
               
               
                 gac tgc att  tat tct gca gac ttt  ggt ggt tct cta aca  tta cag     10820  
               
               
                 Asp Cys Ile  Tyr Ser Ala Asp Phe  Gly Gly Ser Leu Thr  Leu Gln  
               
               
                         3570                 3575                 3580  
               
               
                   
               
               
                 tac aaa gct  gac agg gag gga cat  tgt cca gtt cac tcc  cac tcc     10865  
               
               
                 Tyr Lys Ala  Asp Arg Glu Gly His  Cys Pro Val His Ser  His Ser  
               
               
                         3585                 3590                 3595  
               
               
                   
               
               
                 acg aca gct  gtt ttg aag gaa gcg  acc aca cat gtg act  gcc gta     10910  
               
               
                 Thr Thr Ala  Val Leu Lys Glu Ala  Thr Thr His Val Thr  Ala Val  
               
               
                         3600                 3605                 3610  
               
               
                   
               
               
                 ggc agc ata  aca cta cat ttt agc  aca tcg agc cca caa  gca aat     10955  
               
               
                 Gly Ser Ile  Thr Leu His Phe Ser  Thr Ser Ser Pro Gln  Ala Asn  
               
               
                         3615                 3620                 3625  
               
               
                   
               
               
                 ttt ata gtt  tcg cta tgc ggc aag  aag tcc acc tgc aat  gct gaa     11000  
               
               
                 Phe Ile Val  Ser Leu Cys Gly Lys  Lys Ser Thr Cys Asn  Ala Glu  
               
               
                         3630                 3635                 3640  
               
               
                   
               
               
                 tgt aaa cca  ccg gcc gac cac ata  att gga gaa cca cat  aaa gtc     11045  
               
               
                 Cys Lys Pro  Pro Ala Asp His Ile  Ile Gly Glu Pro His  Lys Val  
               
               
                         3645                 3650                 3655  
               
               
                   
               
               
                 gac caa gaa  ttc cag gcg gca gtt  tcc aaa aca tct tgg  aac tgg     11090  
               
               
                 Asp Gln Glu  Phe Gln Ala Ala Val  Ser Lys Thr Ser Trp  Asn Trp  
               
               
                         3660                 3665                 3670  
               
               
                   
               
               
                 ctg ctt gca  ctg ttt ggg gga gca  tca tcc ctc att gtt  gta gga     11135  
               
               
                 Leu Leu Ala  Leu Phe Gly Gly Ala  Ser Ser Leu Ile Val  Val Gly  
               
               
                         3675                 3680                 3685  
               
               
                   
               
               
                 ctt ata gtg  ttg gtc tgc agc tct  atg ctt ata aac aca  cgt aga     11180  
               
               
                 Leu Ile Val  Leu Val Cys Ser Ser  Met Leu Ile Asn Thr  Arg Arg  
               
               
                         3690                 3695                 3700  
               
               
                   
               
               
                 tga ctgagcgcgg acactgacat agcggtaaaa ctcgatgtac ttccgaggaa         11233  
               
               
                   
               
               
                 gcgtggtgca taatgccacg cgccgcttga cactaaaact cgatgtattt ccgaggaagc  11293  
               
               
                   
               
               
                 acagtgcata atgctgtgca gtgtcacatt aatcgtatat cacactacat attaacaaca  11353  
               
               
                   
               
               
                 ctatatcact tttatgagac tcactatggg tctctaatat acactacaca tattttactt  11413  
               
               
                   
               
               
                 aaaaacacta tacacacttt ataaattctt ttataatttt tcttttgttt ttattttgtt  11473  
               
               
                   
               
               
                 tttaaaattt c                                                       11484  
               
               
                   
               
               
                   
               
               
                 &lt;210&gt; SEQ ID NO 2  
               
               
                 &lt;211&gt; LENGTH: 1852  
               
               
                 &lt;212&gt; TYPE: PRT  
               
               
                 &lt;213&gt; ORGANISM: Western equine encephalomyelitis virus - strain  
               
               
                       71V-1658  
               
               
                   
               
               
                 &lt;400&gt; SEQUENCE: 2  
               
               
                   
               
               
                 Met Glu Arg Ile His Val Asp Leu Asp Ala Asp Ser Pro Tyr Val Lys  
               
               
                 1               5                   10                  15  
               
               
                   
               
               
                 Ser Leu Gln Arg Thr Phe Pro Gln Phe Glu Ile Glu Ala Arg Gln Val  
               
               
                             20                  25                  30  
               
               
                   
               
               
                 Thr Asp Asn Asp His Ala Asn Ala Arg Ala Phe Ser His Val Ala Thr  
               
               
                         35                  40                  45  
               
               
                   
               
               
                 Lys Leu Ile Glu Ser Glu Val Asp Arg Asp Gln Val Ile Leu Asp Ile  
               
               
                     50                  55                  60  
               
               
                   
               
               
                 Gly Ser Ala Pro Val Arg His Ala His Ser Asn His Arg Tyr His Cys  
               
               
                 65                  70                  75                  80  
               
               
                   
               
               
                 Ile Cys Pro Met Ile Ser Ala Glu Asp Pro Asp Arg Leu Gln Arg Tyr  
               
               
                                 85                  90                  95  
               
               
                   
               
               
                 Ala Glu Arg Leu Lys Lys Ser Asp Ile Thr Asp Lys Asn Ile Ala Ser  
               
               
                             100                 105                 110  
               
               
                   
               
               
                 Lys Ala Ala Asp Leu Leu Glu Val Met Ser Thr Pro Asp Ala Glu Thr  
               
               
                         115                 120                 125  
               
               
                   
               
               
                 Pro Ser Leu Cys Met His Thr Asp Ala Thr Cys Arg Tyr Phe Gly Ser  
               
               
                     130                 135                 140  
               
               
                   
               
               
                 Val Ala Val Tyr Gln Asp Val Tyr Ala Val His Ala Pro Thr Ser Ile  
               
               
                 145                 150                 155                 160  
               
               
                   
               
               
                 Tyr His Gln Ala Leu Lys Gly Val Arg Thr Ile Tyr Trp Ile Gly Phe  
               
               
                                 165                 170                 175  
               
               
                   
               
               
                 Asp Thr Thr Pro Phe Met Tyr Lys Asn Met Ala Gly Ser Tyr Pro Thr  
               
               
                             180                 185                 190  
               
               
                   
               
               
                 Tyr Asn Thr Asn Trp Ala Asp Glu Arg Val Leu Glu Ala Arg Asn Ile  
               
               
                         195                 200                 205  
               
               
                   
               
               
                 Gly Leu Gly Asn Ser Asp Leu Gln Glu Ser Arg Leu Gly Lys Leu Ser  
               
               
                     210                 215                 220  
               
               
                   
               
               
                 Ile Leu Arg Lys Lys Arg Leu Gln Pro Thr Asn Lys Ile Ile Phe Ser  
               
               
                 225                 230                 235                 240  
               
               
                   
               
               
                 Val Gly Ser Thr Ile Tyr Thr Glu Asp Arg Ser Leu Leu Arg Ser Trp  
               
               
                                 245                 250                 255  
               
               
                   
               
               
                 His Leu Pro Asn Val Phe His Leu Lys Gly Lys Ser Asn Phe Thr Gly  
               
               
                             260                 265                 270  
               
               
                   
               
               
                 Arg Cys Gly Thr Ile Val Ser Cys Glu Gly Tyr Val Ile Lys Lys Ile  
               
               
                         275                 280                 285  
               
               
                   
               
               
                 Thr Ile Ser Pro Gly Leu Tyr Gly Lys Val Glu Asn Leu Ala Ser Thr  
               
               
                     290                 295                 300  
               
               
                   
               
               
                 Met His Arg Glu Gly Phe Leu Ser Cys Lys Val Thr Asp Thr Leu Arg  
               
               
                 305                 310                 315                 320  
               
               
                   
               
               
                 Gly Glu Arg Val Ser Phe Ala Val Cys Thr Tyr Val Pro Ala Thr Leu  
               
               
                                 325                 330                 335  
               
               
                   
               
               
                 Cys Asp Gln Met Thr Gly Ile Leu Ala Thr Asp Val Ser Val Asp Asp  
               
               
                             340                 345                 350  
               
               
                   
               
               
                 Ala Gln Lys Leu Leu Val Gly Leu Asn Gln Arg Ile Val Val Asn Gly  
               
               
                         355                 360                 365  
               
               
                   
               
               
                 Arg Thr Gln Arg Asn Thr Asn Thr Met Gln Asn Tyr Leu Leu Pro Val  
               
               
                     370                 375                 380  
               
               
                   
               
               
                 Val Ala Gln Ala Phe Ser Arg Trp Ala Arg Glu His Arg Ala Asp Leu  
               
               
                 385                 390                 395                 400  
               
               
                   
               
               
                 Asp Asp Glu Lys Glu Leu Gly Val Arg Glu Arg Thr Leu Thr Met Gly  
               
               
                                 405                 410                 415  
               
               
                   
               
               
                 Cys Cys Trp Ala Phe Lys Thr Gln Lys Ile Thr Ser Ile Tyr Lys Lys  
               
               
                             420                 425                 430  
               
               
                   
               
               
                 Pro Gly Thr Gln Thr Ile Lys Lys Val Pro Ala Val Phe Asp Ser Phe  
               
               
                         435                 440                 445  
               
               
                   
               
               
                 Val Ile Pro Arg Leu Thr Ser His Gly Leu Asp Met Gly Phe Arg Arg  
               
               
                     450                 455                 460  
               
               
                   
               
               
                 Arg Leu Lys Leu Leu Leu Glu Pro Thr Val Lys Pro Ala Pro Ala Ile  
               
               
                 465                 470                 475                 480  
               
               
                   
               
               
                 Thr Met Ala Asp Val Glu His Leu Arg Gly Leu Gln Gln Glu Ala Glu  
               
               
                                 485                 490                 495  
               
               
                   
               
               
                 Glu Val Ala Ala Ala Glu Glu Ile Arg Glu Ala Leu Pro Pro Leu Leu  
               
               
                             500                 505                 510  
               
               
                   
               
               
                 Pro Glu Ile Glu Lys Glu Thr Val Glu Ala Glu Val Asp Leu Ile Met  
               
               
                         515                 520                 525  
               
               
                   
               
               
                 Gln Glu Ala Gly Ala Gly Ser Val Glu Thr Pro Arg Gly His Ile Arg  
               
               
                     530                 535                 540  
               
               
                   
               
               
                 Val Thr Ser Tyr Pro Gly Glu Glu Lys Ile Gly Ser Tyr Ala Ile Leu  
               
               
                 545                 550                 555                 560  
               
               
                   
               
               
                 Ser Pro Gln Ala Val Leu Asn Ser Glu Lys Leu Ala Cys Ile His Pro  
               
               
                                 565                 570                 575  
               
               
                   
               
               
                 Leu Ala Glu Gln Val Leu Val Met Thr His Lys Gly Arg Ala Gly Arg  
               
               
                             580                 585                 590  
               
               
                   
               
               
                 Tyr Lys Val Glu Pro Tyr His Gly Lys Val Ile Val Pro Glu Gly Thr  
               
               
                         595                 600                 605  
               
               
                   
               
               
                 Ala Val Pro Val Gln Asp Phe Gln Ala Leu Ser Glu Ser Ala Thr Ile  
               
               
                     610                 615                 620  
               
               
                   
               
               
                 Val Phe Asn Glu Arg Glu Phe Val Asn Arg Tyr Leu His His Ile Ala  
               
               
                 625                 630                 635                 640  
               
               
                   
               
               
                 Ile Asn Gly Gly Ala Leu Asn Thr Asp Glu Glu Tyr Tyr Lys Thr Val  
               
               
                                 645                 650                 655  
               
               
                   
               
               
                 Lys Thr Gln Asp Thr Asp Ser Glu Tyr Val Phe Asp Ile Asp Ala Arg  
               
               
                             660                 665                 670  
               
               
                   
               
               
                 Lys Cys Val Lys Arg Glu Asp Ala Gly Pro Leu Cys Leu Thr Gly Asp  
               
               
                         675                 680                 685  
               
               
                   
               
               
                 Leu Val Asp Pro Pro Phe His Glu Phe Ala Tyr Glu Ser Leu Lys Thr  
               
               
                     690                 695                 700  
               
               
                   
               
               
                 Arg Pro Ala Ala Pro His Lys Val Pro Thr Ile Gly Val Tyr Gly Val  
               
               
                 705                 710                 715                 720  
               
               
                   
               
               
                 Pro Gly Ser Gly Lys Ser Gly Ile Ile Lys Ser Ala Val Thr Lys Lys  
               
               
                                 725                 730                 735  
               
               
                   
               
               
                 Asp Leu Val Val Ser Ala Lys Lys Glu Asn Cys Ala Glu Ile Ile Arg  
               
               
                             740                 745                 750  
               
               
                   
               
               
                 Asp Val Arg Arg Met Arg Arg Met Asp Val Ala Ala Arg Thr Val Asp  
               
               
                         755                 760                 765  
               
               
                   
               
               
                 Ser Val Leu Leu Asn Gly Val Lys His Pro Val Asn Thr Leu Tyr Ile  
               
               
                     770                 775                 780  
               
               
                   
               
               
                 Asp Glu Ala Phe Ala Cys His Ala Gly Thr Leu Leu Ala Leu Ile Ala  
               
               
                 785                 790                 795                 800  
               
               
                   
               
               
                 Ile Val Lys Pro Lys Lys Val Val Leu Cys Gly Asp Pro Lys Gln Cys  
               
               
                                 805                 810                 815  
               
               
                   
               
               
                 Gly Phe Phe Asn Met Met Cys Leu Lys Val His Phe Asn His Asp Ile  
               
               
                             820                 825                 830  
               
               
                   
               
               
                 Cys Thr Glu Val Tyr His Lys Ser Ile Ser Arg Arg Cys Thr Gln Thr  
               
               
                         835                 840                 845  
               
               
                   
               
               
                 Val Thr Ala Ile Val Ser Thr Leu Phe Tyr Asp Lys Arg Met Lys Thr  
               
               
                     850                 855                 860  
               
               
                   
               
               
                 Val Asn Pro Cys Ala Asp Lys Ile Ile Ile Asp Thr Thr Gly Thr Thr  
               
               
                 865                 870                 875                 880  
               
               
                   
               
               
                 Lys Pro His Lys Asp Asp Leu Ile Leu Thr Cys Phe Arg Gly Trp Val  
               
               
                                 885                 890                 895  
               
               
                   
               
               
                 Lys Gln Leu Gln Ile Asp Tyr Lys Asn His Glu Ile Met Thr Ala Ala  
               
               
                             900                 905                 910  
               
               
                   
               
               
                 Ala Ser Gln Gly Leu Thr Arg Lys Gly Val Tyr Ala Val Arg Tyr Lys  
               
               
                         915                 920                 925  
               
               
                   
               
               
                 Val Asn Glu Asn Pro Leu Tyr Ser Gln Thr Ser Glu His Val Asn Val  
               
               
                     930                 935                 940  
               
               
                   
               
               
                 Leu Leu Thr Arg Thr Glu Lys Arg Ile Val Trp Lys Thr Leu Ala Gly  
               
               
                 945                 950                 955                 960  
               
               
                   
               
               
                 Asp Pro Trp Ile Lys Thr Leu Thr Ala Lys Tyr Pro Gly Asp Phe Thr  
               
               
                                 965                 970                 975  
               
               
                   
               
               
                 Ala Ser Leu Asp Asp Trp Gln Arg Glu His Asp Ala Ile Met Ala Arg  
               
               
                             980                 985                 990  
               
               
                   
               
               
                 Val Leu Asp Lys Pro Gln Thr Ala  Asp Val Phe Gln Asn  Lys Val Asn  
               
               
                         995                 1000                 1005  
               
               
                   
               
               
                 Val Cys  Trp Ala Lys Ala Leu  Glu Pro Val Leu Ala  Thr Ala Asn  
               
               
                     1010                 1015                 1020  
               
               
                   
               
               
                 Ile Val  Leu Thr Arg Gln Gln  Trp Glu Thr Leu His  Pro Phe Lys  
               
               
                     1025                 1030                 1035  
               
               
                   
               
               
                 His Asp  Arg Ala Tyr Ser Pro  Glu Met Ala Leu Asn  Phe Phe Cys  
               
               
                     1040                 1045                 1050  
               
               
                   
               
               
                 Thr Arg  Phe Phe Gly Val Asp  Leu Asp Ser Gly Leu  Phe Ser Ala  
               
               
                     1055                 1060                 1065  
               
               
                   
               
               
                 Pro Thr  Val Ala Leu Thr Tyr  Arg Asp Gln His Trp  Asp Asn Ser  
               
               
                     1070                 1075                 1080  
               
               
                   
               
               
                 Pro Gly  Lys Asn Met Tyr Gly  Leu Asn Arg Glu Val  Ala Lys Glu  
               
               
                     1085                 1090                 1095  
               
               
                   
               
               
                 Leu Ser  Arg Arg Tyr Pro Cys  Ile Thr Lys Ala Val  Asp Thr Gly  
               
               
                     1100                 1105                 1110  
               
               
                   
               
               
                 Arg Val  Ala Asp Ile Arg Asn  Asn Thr Ile Lys Asp  Tyr Ser Pro  
               
               
                     1115                 1120                 1125  
               
               
                   
               
               
                 Thr Ile  Asn Val Val Pro Leu  Asn Arg Arg Leu Pro  His Ser Leu  
               
               
                     1130                 1135                 1140  
               
               
                   
               
               
                 Ile Val  Asp His Lys Gly Gln  Gly Thr Thr Asp His  Ser Gly Phe  
               
               
                     1145                 1150                 1155  
               
               
                   
               
               
                 Leu Ser  Lys Met Lys Gly Lys  Ser Val Leu Val Ile  Gly Asp Pro  
               
               
                     1160                 1165                 1170  
               
               
                   
               
               
                 Ile Ser  Ile Pro Gly Lys Lys  Val Glu Ser Met Gly  Pro Leu Pro  
               
               
                     1175                 1180                 1185  
               
               
                   
               
               
                 Thr Asn  Thr Ile Arg Cys Asp  Leu Asp Leu Gly Ile  Pro Ser His  
               
               
                     1190                 1195                 1200  
               
               
                   
               
               
                 Val Gly  Lys Tyr Asp Ile Ile  Phe Val Asn Val Arg  Thr Pro Tyr  
               
               
                     1205                 1210                 1215  
               
               
                   
               
               
                 Arg Asn  His His Tyr Gln Gln  Cys Glu Asp His Ala  Ile His His  
               
               
                     1220                 1225                 1230  
               
               
                   
               
               
                 Ser Met  Leu Thr Cys Lys Ala  Val His His Leu Asn  Thr Gly Gly  
               
               
                     1235                 1240                 1245  
               
               
                   
               
               
                 Thr Cys  Val Ala Ile Gly Tyr  Gly Leu Ala Asp Arg  Ala Thr Glu  
               
               
                     1250                 1255                 1260  
               
               
                   
               
               
                 Asn Ile  Ile Thr Ala Val Ala  Arg Ser Phe Arg Phe  Thr Arg Val  
               
               
                     1265                 1270                 1275  
               
               
                   
               
               
                 Cys Gln  Pro Lys Asn Thr Ala  Glu Asn Thr Glu Val  Leu Phe Val  
               
               
                     1280                 1285                 1290  
               
               
                   
               
               
                 Phe Phe  Gly Lys Asp Asn Gly  Asn His Thr His Asp  Gln Asp Arg  
               
               
                     1295                 1300                 1305  
               
               
                   
               
               
                 Leu Gly  Val Val Leu Asp Asn  Ile Tyr Gln Gly Ser  Thr Arg Tyr  
               
               
                     1310                 1315                 1320  
               
               
                   
               
               
                 Glu Ala  Gly Arg Ala Pro Ala  Tyr Arg Val Ile Arg  Gly Asp Ile  
               
               
                     1325                 1330                 1335  
               
               
                   
               
               
                 Ser Lys  Ser Ala Asp Gln Ala  Ile Val Asn Ala Ala  Asn Ser Lys  
               
               
                     1340                 1345                 1350  
               
               
                   
               
               
                 Gly Gln  Pro Gly Ser Gly Val  Cys Gly Ala Leu Tyr  Arg Lys Trp  
               
               
                     1355                 1360                 1365  
               
               
                   
               
               
                 Pro Ala  Ala Phe Asp Arg Gln  Pro Ile Ala Val Gly  Thr Ala Arg  
               
               
                     1370                 1375                 1380  
               
               
                   
               
               
                 Leu Val  Lys His Glu Pro Leu  Ile Ile His Ala Val  Gly Pro Asn  
               
               
                     1385                 1390                 1395  
               
               
                   
               
               
                 Phe Ser  Lys Met Pro Glu Pro  Glu Gly Asp Leu Lys  Leu Ala Ala  
               
               
                     1400                 1405                 1410  
               
               
                   
               
               
                 Ala Tyr  Met Ser Ile Ala Ser  Ile Val Asn Ala Glu  Arg Ile Thr  
               
               
                     1415                 1420                 1425  
               
               
                   
               
               
                 Lys Ile  Ser Val Pro Leu Leu  Ser Thr Gly Ile Tyr  Ser Gly Gly  
               
               
                     1430                 1435                 1440  
               
               
                   
               
               
                 Lys Asp  Arg Val Met Gln Ser  Leu His His Leu Phe  Thr Ala Phe  
               
               
                     1445                 1450                 1455  
               
               
                   
               
               
                 Asp Thr  Thr Asp Ala Asp Val  Thr Ile Tyr Cys Leu  Asp Lys Gln  
               
               
                     1460                 1465                 1470  
               
               
                   
               
               
                 Trp Glu  Thr Arg Ile Ile Glu  Ala Ile His Arg Lys  Glu Ser Val  
               
               
                     1475                 1480                 1485  
               
               
                   
               
               
                 Glu Ile  Leu Asp Asp Asp Lys  Pro Val Asp Ile Asp  Leu Val Arg  
               
               
                     1490                 1495                 1500  
               
               
                   
               
               
                 Val His  Pro Asn Ser Ser Leu  Ala Gly Arg Pro Gly  Tyr Ser Val  
               
               
                     1505                 1510                 1515  
               
               
                   
               
               
                 Asn Glu  Gly Lys Leu Tyr Ser  Tyr Leu Glu Gly Thr  Arg Phe His  
               
               
                     1520                 1525                 1530  
               
               
                   
               
               
                 Gln Thr  Ala Lys Asp Ile Ala  Glu Ile His Ala Met  Trp Pro Asn  
               
               
                     1535                 1540                 1545  
               
               
                   
               
               
                 Lys Ser  Glu Ala Asn Glu Gln  Ile Cys Leu Tyr Ile  Leu Gly Glu  
               
               
                     1550                 1555                 1560  
               
               
                   
               
               
                 Ser Met  Ser Ser Ile Arg Ser  Lys Cys Pro Val Glu  Glu Ser Glu  
               
               
                     1565                 1570                 1575  
               
               
                   
               
               
                 Ala Ser  Ala Pro Pro His Thr  Leu Pro Cys Leu Cys  Asn Tyr Ala  
               
               
                     1580                 1585                 1590  
               
               
                   
               
               
                 Met Thr  Ala Glu Arg Val Tyr  Arg Leu Arg Ser Ala  Lys Lys Glu  
               
               
                     1595                 1600                 1605  
               
               
                   
               
               
                 Gln Phe  Ala Val Cys Ser Ser  Phe Leu Leu Pro Lys  Tyr Arg Ile  
               
               
                     1610                 1615                 1620  
               
               
                   
               
               
                 Thr Gly  Val Gln Lys Leu Gln  Cys Ser Lys Pro Val  Leu Phe Ser  
               
               
                     1625                 1630                 1635  
               
               
                   
               
               
                 Gly Val  Val Pro Pro Ala Val  His Pro Arg Lys Tyr  Ala Glu Ile  
               
               
                     1640                 1645                 1650  
               
               
                   
               
               
                 Ile Leu  Glu Thr Pro Pro Pro  Pro Ala Thr Thr Thr  Val Ile Cys  
               
               
                     1655                 1660                 1665  
               
               
                   
               
               
                 Glu Pro  Thr Val Pro Glu Arg  Ile Pro Ser Pro Val  Ile Ser Arg  
               
               
                     1670                 1675                 1680  
               
               
                   
               
               
                 Ala Pro  Ser Ala Glu Ser Leu  Leu Ser Leu Gly Gly  Val Ser Phe  
               
               
                     1685                 1690                 1695  
               
               
                   
               
               
                 Ser Ser  Ser Ala Thr Arg Ser  Ser Thr Ala Trp Ser  Asp Tyr Asp  
               
               
                     1700                 1705                 1710  
               
               
                   
               
               
                 Arg Arg  Phe Val Val Thr Ala  Asp Val His Gln Ala  Asn Thr Ser  
               
               
                     1715                 1720                 1725  
               
               
                   
               
               
                 Thr Trp  Ser Ile Pro Ser Ala  Pro Gly Leu Asp Val  Gln Leu Pro  
               
               
                     1730                 1735                 1740  
               
               
                   
               
               
                 Ser Asp  Val Thr Asp Ser His  Trp Ser Ile Pro Ser  Ala Ser Gly  
               
               
                     1745                 1750                 1755  
               
               
                   
               
               
                 Phe Glu  Val Arg Thr Pro Ser  Val Gln Asp Leu Thr  Ala Glu Cys  
               
               
                     1760                 1765                 1770  
               
               
                   
               
               
                 Ala Lys  Pro Arg Gly Leu Ala  Glu Ile Met Gln Asp  Phe Asn Thr  
               
               
                     1775                 1780                 1785  
               
               
                   
               
               
                 Ala Pro  Phe Gln Phe Leu Ser  Asp Tyr Arg Pro Val  Pro Ala Pro  
               
               
                     1790                 1795                 1800  
               
               
                   
               
               
                 Arg Arg  Arg Pro Ile Pro Ser  Pro Arg Ser Thr Ala  Ser Ala Pro  
               
               
                     1805                 1810                 1815  
               
               
                   
               
               
                 Pro Val  Pro Lys Pro Arg Arg  Thr Lys Tyr Gln Gln  Pro Pro Gly  
               
               
                     1820                 1825                 1830  
               
               
                   
               
               
                 Val Ala  Arg Ala Ile Ser Glu  Ala Glu Leu Asp Glu  Tyr Ile Arg  
               
               
                     1835                 1840                 1845  
               
               
                   
               
               
                 Gln His  Ser Asn  
               
               
                     1850  
               
               
                   
               
               
                   
               
               
                 &lt;210&gt; SEQ ID NO 3  
               
               
                 &lt;211&gt; LENGTH: 614  
               
               
                 &lt;212&gt; TYPE: PRT  
               
               
                 &lt;213&gt; ORGANISM: Western equine encephalomyelitis virus - strain 71V-1658  
               
               
                   
               
               
                 &lt;400&gt; SEQUENCE: 3  
               
               
                   
               
               
                 Arg Tyr Glu Ala Gly Ala Tyr Ile Phe Ser Ser Glu Thr Gly Gln Gly  
               
               
                 1               5                   10                  15  
               
               
                   
               
               
                 His Leu Gln Gln Lys Ser Val Arg Gln Cys Lys Leu Gln Glu Pro Ile  
               
               
                             20                  25                  30  
               
               
                   
               
               
                 Leu Asp Arg Ala Val His Glu Lys Tyr Tyr Ala Pro Arg Leu Asp Leu  
               
               
                         35                  40                  45  
               
               
                   
               
               
                 Glu Arg Glu Lys Met Leu Gln Lys Lys Leu Gln Leu Cys Ala Ser Glu  
               
               
                     50                  55                  60  
               
               
                   
               
               
                 Gly Asn Arg Ser Arg Tyr Gln Ser Arg Lys Val Glu Asn Met Lys Ala  
               
               
                 65                  70                  75                  80  
               
               
                   
               
               
                 Ile Thr Ala Glu Arg Leu Ile Ser Gly Leu Gly Thr Tyr Leu Ser Ser  
               
               
                                 85                  90                  95  
               
               
                   
               
               
                 Glu Val Asn Pro Val Glu Cys Tyr Arg Val Asn Tyr Pro Val Pro Ile  
               
               
                             100                 105                 110  
               
               
                   
               
               
                 Tyr Ser Ser Thr Val Ile Asn Arg Phe Thr Ser Ala Glu Val Ala Val  
               
               
                         115                 120                 125  
               
               
                   
               
               
                 Lys Thr Cys Asn Leu Val Ile Gln Glu Asn Tyr Pro Thr Val Ala Ser  
               
               
                     130                 135                 140  
               
               
                   
               
               
                 Tyr Cys Ile Thr Asp Glu Tyr Asp Ala Tyr Leu Asp Met Val Asp Gly  
               
               
                 145                 150                 155                 160  
               
               
                   
               
               
                 Ala Ser Cys Cys Leu Asp Thr Ala Thr Phe Cys Pro Ala Lys Leu Arg  
               
               
                                 165                 170                 175  
               
               
                   
               
               
                 Ser Tyr Pro Lys Lys His Ser Tyr Leu Gln Pro Glu Ile Arg Ser Ala  
               
               
                             180                 185                 190  
               
               
                   
               
               
                 Val Pro Ser Pro Ile Gln Asn Thr Leu Gln Asn Val Leu Ala Ala Ala  
               
               
                         195                 200                 205  
               
               
                   
               
               
                 Thr Lys Arg Asn Cys Asn Val Thr Gln Met Arg Glu Leu Pro Val Leu  
               
               
                     210                 215                 220  
               
               
                   
               
               
                 Asp Ser Ala Ala Phe Asn Val Asp Cys Phe Lys Lys Tyr Ala Cys Asn  
               
               
                 225                 230                 235                 240  
               
               
                   
               
               
                 Asp Glu Tyr Trp Asp Thr Phe Arg Asp Asn Pro Ile Arg Leu Thr Thr  
               
               
                                 245                 250                 255  
               
               
                   
               
               
                 Glu Asn Val Thr Gln Tyr Val Thr Lys Leu Lys Gly Pro Lys Ala Ala  
               
               
                             260                 265                 270  
               
               
                   
               
               
                 Ala Leu Phe Ala Asn Thr His Asn Leu Lys Pro Leu Gln Glu Ile Pro  
               
               
                         275                 280                 285  
               
               
                   
               
               
                 Met Asp Gln Phe Val Met Asp Leu Lys Arg Asp Val Lys Val Thr Pro  
               
               
                     290                 295                 300  
               
               
                   
               
               
                 Gly Thr Lys His Thr Glu Glu Arg Pro Lys Val Gln Val Ile Gln Ala  
               
               
                 305                 310                 315                 320  
               
               
                   
               
               
                 Ala Asp Pro Leu Ala Thr Ala Tyr Leu Cys Gly Ile His Arg Glu Leu  
               
               
                                 325                 330                 335  
               
               
                   
               
               
                 Val Arg Arg Leu Asn Ala Val Leu Leu Pro Asn Ile His Thr Leu Phe  
               
               
                             340                 345                 350  
               
               
                   
               
               
                 Asp Met Ser Ala Glu Asp Phe Asp Ala Ile Ile Ala Glu His Phe His  
               
               
                         355                 360                 365  
               
               
                   
               
               
                 His Gly Asp Pro Val Leu Glu Thr Asp Ile Ala Ser Phe Asp Lys Ser  
               
               
                     370                 375                 380  
               
               
                   
               
               
                 Glu Asp Asp Ala Ile Ala Ile Ser Ala Leu Met Ile Leu Glu Asp Leu  
               
               
                 385                 390                 395                 400  
               
               
                   
               
               
                 Gly Val Asp Gln Pro Leu Leu Asp Leu Ile Glu Ala Ala Phe Gly Asn  
               
               
                                 405                 410                 415  
               
               
                   
               
               
                 Ile Thr Ser Val His Leu Pro Thr Gly Thr Arg Phe Lys Phe Gly Ala  
               
               
                             420                 425                 430  
               
               
                   
               
               
                 Met Met Lys Ser Gly Met Phe Leu Thr Leu Phe Val Asn Thr Leu Val  
               
               
                         435                 440                 445  
               
               
                   
               
               
                 Asn Ile Met Ile Ala Ser Arg Val Leu Arg Glu Arg Leu Thr Thr Ser  
               
               
                     450                 455                 460  
               
               
                   
               
               
                 Ala Cys Ala Ala Ser Ile Gly Asp Asp Asn Ile Val His Gly Val Val  
               
               
                 465                 470                 475                 480  
               
               
                   
               
               
                 Ser Asp Thr Leu Met Ala Glu Arg Cys Ala Thr Trp Leu Asn Met Glu  
               
               
                                 485                 490                 495  
               
               
                   
               
               
                 Val Lys Ile Ile Asp Ala Val Ile Gly Ile Lys Ala Pro Tyr Phe Cys  
               
               
                             500                 505                 510  
               
               
                   
               
               
                 Gly Gly Phe Ile Leu Val Asp Gln Ile Thr Gly Thr Ala Cys Arg Val  
               
               
                         515                 520                 525  
               
               
                   
               
               
                 Ala Asp Pro Leu Lys Arg Leu Phe Lys Leu Gly Lys Pro Leu Pro Val  
               
               
                     530                 535                 540  
               
               
                   
               
               
                 Asp Asp Thr Gln Asp Cys Asp Arg Arg Arg Ala Leu His Asp Glu Ala  
               
               
                 545                 550                 555                 560  
               
               
                   
               
               
                 Met Arg Trp Asn Arg Ile Gly Ile Thr Asp Glu Leu Val Lys Ala Val  
               
               
                                 565                 570                 575  
               
               
                   
               
               
                 Glu Ser Arg Tyr Glu Ile Ile Leu Ala Gly Leu Ile Ile Thr Ser Leu  
               
               
                             580                 585                 590  
               
               
                   
               
               
                 Ser Thr Leu Ala Glu Ser Val Lys Asn Phe Lys Ser Ile Arg Gly Ser  
               
               
                         595                 600                 605  
               
               
                   
               
               
                 Pro Ile Thr Leu Tyr Gly  
               
               
                     610  
               
               
                   
               
               
                   
               
               
                 &lt;210&gt; SEQ ID NO 4  
               
               
                 &lt;211&gt; LENGTH: 1236  
               
               
                 &lt;212&gt; TYPE: PRT  
               
               
                 &lt;213&gt; ORGANISM: Western equine encephalomyelitis virus - strain 71V-1658  
               
               
                   
               
               
                 &lt;400&gt; SEQUENCE: 4  
               
               
                   
               
               
                 Met Phe Pro Tyr Pro Gln Leu Asn Phe Pro Pro Val Tyr Pro Thr Asn  
               
               
                 1               5                   10                  15  
               
               
                   
               
               
                 Pro Met Ala Tyr Arg Asp Pro Asn Pro Pro Arg Arg Arg Trp Arg Pro  
               
               
                             20                  25                  30  
               
               
                   
               
               
                 Phe Arg Pro Pro Leu Ala Ala Gln Ile Glu Asp Leu Arg Arg Ser Ile  
               
               
                         35                  40                  45  
               
               
                   
               
               
                 Val Asn Leu Thr Phe Lys Gln Arg Ser Pro Asn Pro Pro Pro Gly Pro  
               
               
                     50                  55                  60  
               
               
                   
               
               
                 Pro Pro Lys Lys Lys Lys Ser Ala Pro Lys Pro Lys Pro Thr Gln Pro  
               
               
                 65                  70                  75                  80  
               
               
                   
               
               
                 Lys Lys Lys Lys Gln Gln Ala Lys Arg Thr Lys Arg Lys Pro Lys Pro  
               
               
                                 85                  90                  95  
               
               
                   
               
               
                 Gly Lys Arg Gln Arg Met Cys Met Lys Leu Glu Ser Asp Lys Thr Phe  
               
               
                             100                 105                 110  
               
               
                   
               
               
                 Pro Ile Met Leu Asn Gly Gln Val Asn Gly Tyr Ala Cys Val Val Gly  
               
               
                         115                 120                 125  
               
               
                   
               
               
                 Gly Arg Leu Met Lys Pro Leu His Val Glu Gly Lys Ile Asp Asn Glu  
               
               
                     130                 135                 140  
               
               
                   
               
               
                 Gln Leu Ala Ala Val Lys Leu Lys Lys Ala Ser Met Tyr Asp Leu Glu  
               
               
                 145                 150                 155                 160  
               
               
                   
               
               
                 Tyr Gly Asp Val Pro Gln Asn Met Lys Ser Asp Thr Leu Gln Tyr Thr  
               
               
                                 165                 170                 175  
               
               
                   
               
               
                 Ser Asp Lys Pro Pro Gly Phe Tyr Asn Trp His His Gly Ala Val Gln  
               
               
                             180                 185                 190  
               
               
                   
               
               
                 Tyr Glu Asn Gly Arg Phe Thr Val Pro Arg Gly Val Gly Gly Lys Gly  
               
               
                         195                 200                 205  
               
               
                   
               
               
                 Asp Ser Gly Arg Pro Ile Leu Asp Asn Arg Gly Arg Val Val Ala Ile  
               
               
                     210                 215                 220  
               
               
                   
               
               
                 Val Leu Gly Gly Ala Asn Glu Gly Thr Arg Thr Ala Leu Ser Val Val  
               
               
                 225                 230                 235                 240  
               
               
                   
               
               
                 Thr Trp Asn Gln Lys Gly Val Thr Ile Arg Asp Thr Pro Glu Gly Ser  
               
               
                                 245                 250                 255  
               
               
                   
               
               
                 Glu Pro Trp Ser Leu Val Thr Ala Leu Cys Val Leu Ser Asn Val Thr  
               
               
                             260                 265                 270  
               
               
                   
               
               
                 Phe Pro Cys Asp Lys Pro Pro Val Cys Tyr Ser Leu Thr Pro Glu Arg  
               
               
                         275                 280                 285  
               
               
                   
               
               
                 Thr Leu Asp Val Leu Glu Glu Asn Val Asp Asn Pro Asn Tyr Asp Thr  
               
               
                     290                 295                 300  
               
               
                   
               
               
                 Leu Leu Glu Asn Val Leu Lys Cys Pro Ser Arg Arg Pro Lys Arg Ser  
               
               
                 305                 310                 315                 320  
               
               
                   
               
               
                 Ile Thr Asp Asp Phe Thr Leu Thr Ser Pro Tyr Leu Gly Phe Cys Pro  
               
               
                                 325                 330                 335  
               
               
                   
               
               
                 Tyr Cys Arg His Ser Thr Pro Cys Phe Ser Pro Ile Lys Ile Glu Asn  
               
               
                             340                 345                 350  
               
               
                   
               
               
                 Val Trp Asp Glu Ser Asp Asp Gly Ser Ile Arg Ile Gln Val Ser Ala  
               
               
                         355                 360                 365  
               
               
                   
               
               
                 Gln Phe Gly Tyr Asn Gln Ala Gly Thr Ala Asp Val Thr Lys Phe Arg  
               
               
                     370                 375                 380  
               
               
                   
               
               
                 Tyr Met Ser Phe Asp His Asp His Asp Ile Lys Glu Asp Ser Met Glu  
               
               
                 385                 390                 395                 400  
               
               
                   
               
               
                 Lys Ile Ala Ile Ser Thr Ser Gly Pro Cys Arg Arg Leu Gly His Lys  
               
               
                                 405                 410                 415  
               
               
                   
               
               
                 Gly Tyr Phe Leu Leu Ala Gln Cys Pro Pro Gly Asp Ser Val Thr Val  
               
               
                             420                 425                 430  
               
               
                   
               
               
                 Ser Ile Thr Ser Gly Ala Ser Glu Asn Ser Cys Thr Val Glu Lys Lys  
               
               
                         435                 440                 445  
               
               
                   
               
               
                 Ile Arg Arg Lys Phe Val Gly Arg Glu Glu Tyr Leu Phe Pro Pro Val  
               
               
                     450                 455                 460  
               
               
                   
               
               
                 His Gly Lys Leu Val Lys Cys His Val Tyr Asp His Leu Lys Glu Thr  
               
               
                 465                 470                 475                 480  
               
               
                   
               
               
                 Ser Ala Gly Tyr Ile Thr Met His Arg Pro Gly Pro His Ala Tyr Lys  
               
               
                                 485                 490                 495  
               
               
                   
               
               
                 Ser Tyr Leu Glu Glu Ala Ser Gly Glu Val Tyr Ile Lys Pro Pro Ser  
               
               
                             500                 505                 510  
               
               
                   
               
               
                 Gly Lys Asn Val Thr Tyr Glu Cys Lys Cys Gly Asp Tyr Ser Thr Gly  
               
               
                         515                 520                 525  
               
               
                   
               
               
                 Ile Val Ser Thr Arg Thr Lys Met Asn Gly Cys Thr Lys Ala Lys Gln  
               
               
                     530                 535                 540  
               
               
                   
               
               
                 Cys Ile Ala Tyr Lys Ser Asp Gln Thr Lys Trp Val Phe Asn Ser Pro  
               
               
                 545                 550                 555                 560  
               
               
                   
               
               
                 Asp Leu Ile Arg His Thr Asp His Ser Val Gln Gly Lys Leu His Ile  
               
               
                                 565                 570                 575  
               
               
                   
               
               
                 Pro Phe Arg Leu Thr Pro Thr Val Cys Pro Val Pro Leu Ala His Thr  
               
               
                             580                 585                 590  
               
               
                   
               
               
                 Pro Thr Val Thr Lys Trp Phe Lys Gly Ile Thr Leu His Leu Thr Ala  
               
               
                         595                 600                 605  
               
               
                   
               
               
                 Met Arg Pro Thr Leu Leu Thr Thr Arg Lys Leu Gly Leu Arg Ala Asp  
               
               
                     610                 615                 620  
               
               
                   
               
               
                 Ala Thr Ala Glu Trp Ile Thr Gly Ser Thr Ser Arg Asn Phe Ser Val  
               
               
                 625                 630                 635                 640  
               
               
                   
               
               
                 Gly Arg Glu Gly Leu Glu Tyr Val Trp Gly Asn His Glu Pro Val Arg  
               
               
                                 645                 650                 655  
               
               
                   
               
               
                 Val Trp Ala Gln Glu Ser Ala Pro Gly Asp Pro His Gly Trp Pro His  
               
               
                             660                 665                 670  
               
               
                   
               
               
                 Glu Ile Ile Ile His Tyr Tyr His Arg His Pro Val Tyr Thr Val Ile  
               
               
                         675                 680                 685  
               
               
                   
               
               
                 Val Leu Cys Gly Val Ala Leu Ala Ile Leu Val Gly Thr Ala Ser Ser  
               
               
                     690                 695                 700  
               
               
                   
               
               
                 Ala Ala Cys Ile Ala Lys Ala Arg Arg Asp Cys Leu Thr Pro Tyr Ala  
               
               
                 705                 710                 715                 720  
               
               
                   
               
               
                 Leu Ala Pro Asn Ala Thr Val Pro Thr Ala Leu Ala Val Leu Cys Cys  
               
               
                                 725                 730                 735  
               
               
                   
               
               
                 Ile Arg Pro Thr Asn Ala Glu Thr Phe Gly Glu Thr Leu Asn His Leu  
               
               
                             740                 745                 750  
               
               
                   
               
               
                 Trp Phe Asn Asn Gln Pro Phe Leu Trp Ala Gln Leu Cys Ile Pro Leu  
               
               
                         755                 760                 765  
               
               
                   
               
               
                 Ala Ala Leu Val Ile Leu Phe Arg Cys Phe Ser Cys Cys Met Pro Phe  
               
               
                     770                 775                 780  
               
               
                   
               
               
                 Leu Leu Val Ala Gly Val Cys Leu Gly Lys Val Asp Ala Phe Glu His  
               
               
                 785                 790                 795                 800  
               
               
                   
               
               
                 Ala Thr Thr Val Pro Asn Val Pro Gly Ile Pro Tyr Lys Ala Leu Val  
               
               
                                 805                 810                 815  
               
               
                   
               
               
                 Glu Arg Ala Gly Tyr Ala Pro Leu Asn Leu Glu Ile Thr Val Val Ser  
               
               
                             820                 825                 830  
               
               
                   
               
               
                 Ser Glu Leu Thr Pro Ser Thr Asn Lys Glu Tyr Val Thr Cys Lys Phe  
               
               
                         835                 840                 845  
               
               
                   
               
               
                 His Thr Val Ile Pro Ser Pro Gln Val Lys Cys Cys Gly Ser Leu Glu  
               
               
                     850                 855                 860  
               
               
                   
               
               
                 Cys Lys Ala Ser Ser Lys Ala Asp Tyr Thr Cys Arg Val Phe Gly Gly  
               
               
                 865                 870                 875                 880  
               
               
                   
               
               
                 Val Tyr Pro Phe Met Trp Gly Gly Ala Gln Cys Phe Cys Asp Ser Glu  
               
               
                                 885                 890                 895  
               
               
                   
               
               
                 Asn Thr Gln Leu Ser Glu Ala Tyr Val Glu Phe Ala Pro Asp Cys Thr  
               
               
                             900                 905                 910  
               
               
                   
               
               
                 Ile Asp His Ala Val Ala Leu Lys Val His Thr Ala Ala Leu Lys Val  
               
               
                         915                 920                 925  
               
               
                   
               
               
                 Gly Leu Arg Ile Val Tyr Gly Asn Thr Thr Ala His Leu Asp Thr Phe  
               
               
                     930                 935                 940  
               
               
                   
               
               
                 Val Asn Gly Val Thr Pro Gly Ser Ser Arg Asp Leu Lys Val Ile Ala  
               
               
                 945                 950                 955                 960  
               
               
                   
               
               
                 Gly Pro Ile Ser Ala Ala Phe Ser Pro Phe Asp His Lys Val Val Ile  
               
               
                                 965                 970                 975  
               
               
                   
               
               
                 Arg Lys Gly Leu Val Tyr Asn Tyr Asp Phe Pro Glu Tyr Gly Ala Met  
               
               
                             980                 985                 990  
               
               
                   
               
               
                 Lys Pro Gly Ala Phe Gly Asp Ile  Gln Ala Ser Ser Leu  Asp Ala Thr  
               
               
                         995                 1000                 1005  
               
               
                   
               
               
                 Asp Ile  Val Ala Arg Thr Asp  Ile Arg Leu Leu Lys  Pro Ser Val  
               
               
                     1010                 1015                 1020  
               
               
                   
               
               
                 Lys Asn  Ile His Val Pro Tyr  Thr Gln Ala Val Ser  Gly Tyr Glu  
               
               
                     1025                 1030                 1035  
               
               
                   
               
               
                 Met Trp  Lys Asn Asn Ser Gly  Arg Pro Leu Gln Glu  Thr Ala Pro  
               
               
                     1040                 1045                 1050  
               
               
                   
               
               
                 Phe Gly  Cys Lys Ile Glu Val  Glu Pro Leu Arg Ala  Ser Asn Cys  
               
               
                     1055                 1060                 1065  
               
               
                   
               
               
                 Ala Tyr  Gly His Ile Pro Ile  Ser Ile Asp Ile Pro  Asp Ala Ala  
               
               
                     1070                 1075                 1080  
               
               
                   
               
               
                 Phe Val  Arg Ser Ser Glu Ser  Pro Thr Ile Leu Glu  Val Ser Cys  
               
               
                     1085                 1090                 1095  
               
               
                   
               
               
                 Thr Val  Ala Asp Cys Ile Tyr  Ser Ala Asp Phe Gly  Gly Ser Leu  
               
               
                     1100                 1105                 1110  
               
               
                   
               
               
                 Thr Leu  Gln Tyr Lys Ala Asp  Arg Glu Gly His Cys  Pro Val His  
               
               
                     1115                 1120                 1125  
               
               
                   
               
               
                 Ser His  Ser Thr Thr Ala Val  Leu Lys Glu Ala Thr  Thr His Val  
               
               
                     1130                 1135                 1140  
               
               
                   
               
               
                 Thr Ala  Val Gly Ser Ile Thr  Leu His Phe Ser Thr  Ser Ser Pro  
               
               
                     1145                 1150                 1155  
               
               
                   
               
               
                 Gln Ala  Asn Phe Ile Val Ser  Leu Cys Gly Lys Lys  Ser Thr Cys  
               
               
                     1160                 1165                 1170  
               
               
                   
               
               
                 Asn Ala  Glu Cys Lys Pro Pro  Ala Asp His Ile Ile  Gly Glu Pro  
               
               
                     1175                 1180                 1185  
               
               
                   
               
               
                 His Lys  Val Asp Gln Glu Phe  Gln Ala Ala Val Ser  Lys Thr Ser  
               
               
                     1190                 1195                 1200  
               
               
                   
               
               
                 Trp Asn  Trp Leu Leu Ala Leu  Phe Gly Gly Ala Ser  Ser Leu Ile  
               
               
                     1205                 1210                 1215  
               
               
                   
               
               
                 Val Val  Gly Leu Ile Val Leu  Val Cys Ser Ser Met  Leu Ile Asn  
               
               
                     1220                 1225                 1230  
               
               
                   
               
               
                 Thr Arg  Arg  
               
               
                     1235  
               
               
                   
               
               
                   
               
               
                 &lt;210&gt; SEQ ID NO 5  
               
               
                 &lt;211&gt; LENGTH: 4150  
               
               
                 &lt;212&gt; TYPE: DNA  
               
               
                 &lt;213&gt; ORGANISM: Western equine encephalomyelitis virus - strain 71V-1658  
               
               
                 &lt;220&gt; FEATURE:  
               
               
                 &lt;221&gt; NAME/KEY: CDS  
               
               
                 &lt;222&gt; LOCATION: (159)..(3869)  
               
               
                 &lt;223&gt; OTHER INFORMATION: vector sequence 1-9; 5′ Sacl primer 9-20; 3′ 
               
               
                       end of NS4 gene 16-114; intragenic region 115-158; polyprotein  
               
               
                       (C-E3-E2-6K-E1) 159-3856; pcDW-XH7 nontranslated region 3857-4150  
               
               
                   
               
               
                 &lt;400&gt; SEQUENCE: 5  
               
               
                   
               
               
                 ggccctctag agctcatact ggcaggcctg atcatcacgt ctctgtccac gttagccgaa     60  
               
               
                   
               
               
                 agcgttaaga acttcaagag cataagaggg agcccaatca ccctctacgg ctgacctaaa    120  
               
               
                   
               
               
                 taggtgacgt agtagacacg cacctaccca ccggcaga atg ttt cca tac cct cag    176  
               
               
                                                           Met Phe Pro Tyr Pro Gln  
               
               
                                                           1               5  
               
               
                   
               
               
                 ctg aac ttt cca cca gtt tac cct aca aat ccg atg gct tac cga gat      224  
               
               
                 Leu Asn Phe Pro Pro Val Tyr Pro Thr Asn Pro Met Ala Tyr Arg Asp  
               
               
                             10                  15                  20  
               
               
                   
               
               
                 cca aac cct cct agg cgc cgc tgg agg ccg ttt cgg ccc ccg ctg gct      272  
               
               
                 Pro Asn Pro Pro Arg Arg Arg Trp Arg Pro Phe Arg Pro Pro Leu Ala  
               
               
                         25                  30                  35  
               
               
                   
               
               
                 gct caa atc gaa gat ctt agg agg tcg ata gtc aac ttg act ttc aaa      320  
               
               
                 Ala Gln Ile Glu Asp Leu Arg Arg Ser Ile Val Asn Leu Thr Phe Lys  
               
               
                     40                  45                  50  
               
               
                   
               
               
                 caa cga tca cct aat ccg ccg cca ggt cca ccg cca aag aag aag aag      368  
               
               
                 Gln Arg Ser Pro Asn Pro Pro Pro Gly Pro Pro Pro Lys Lys Lys Lys  
               
               
                 55                  60                  65                  70  
               
               
                   
               
               
                 agt gct cct aag cca aaa cct act cag cct aaa aag aag aag cag caa      416  
               
               
                 Ser Ala Pro Lys Pro Lys Pro Thr Gln Pro Lys Lys Lys Lys Gln Gln  
               
               
                                 75                  80                  85  
               
               
                   
               
               
                 gcc aag agg acg aaa cgc aag cct aaa cca ggg aaa cga caa cgt atg      464  
               
               
                 Ala Lys Arg Thr Lys Arg Lys Pro Lys Pro Gly Lys Arg Gln Arg Met  
               
               
                             90                  95                  100  
               
               
                   
               
               
                 tgt atg aag ttg gag tcg gac aag aca ttt ccg atc atg ctg aac ggc      512  
               
               
                 Cys Met Lys Leu Glu Ser Asp Lys Thr Phe Pro Ile Met Leu Asn Gly  
               
               
                         105                 110                 115  
               
               
                   
               
               
                 caa gtg aat gga tat gcc tgc gtt gtc gga gga agg ctg atg aaa cca      560  
               
               
                 Gln Val Asn Gly Tyr Ala Cys Val Val Gly Gly Arg Leu Met Lys Pro  
               
               
                     120                 125                 130  
               
               
                   
               
               
                 ctc cac gtt gaa gga aaa att gat aat gag caa tta gcg gcc gtg aaa      608  
               
               
                 Leu His Val Glu Gly Lys Ile Asp Asn Glu Gln Leu Ala Ala Val Lys  
               
               
                 135                 140                 145                 150  
               
               
                   
               
               
                 ttg aag aag gct agc atg tac gac ttg gag tac ggc gac gtt ccc cag      656  
               
               
                 Leu Lys Lys Ala Ser Met Tyr Asp Leu Glu Tyr Gly Asp Val Pro Gln  
               
               
                                 155                 160                 165  
               
               
                   
               
               
                 aac atg aaa tca gac acg ctg cag tac acc agc gac aaa cca ccg ggc      704  
               
               
                 Asn Met Lys Ser Asp Thr Leu Gln Tyr Thr Ser Asp Lys Pro Pro Gly  
               
               
                             170                 175                 180  
               
               
                   
               
               
                 ttc tac aac tgg cac cac ggc gca gtc cag tat gag aat ggg aga ttt      752  
               
               
                 Phe Tyr Asn Trp His His Gly Ala Val Gln Tyr Glu Asn Gly Arg Phe  
               
               
                         185                 190                 195  
               
               
                   
               
               
                 acc gta ccg aga gga gtg ggc ggg aaa ggc gac agc gga aga ccg atc      800  
               
               
                 Thr Val Pro Arg Gly Val Gly Gly Lys Gly Asp Ser Gly Arg Pro Ile  
               
               
                     200                 205                 210  
               
               
                   
               
               
                 ctg gac aac aga ggc aga gtt gtg gct att gtt cta gga ggt gca aat      848  
               
               
                 Leu Asp Asn Arg Gly Arg Val Val Ala Ile Val Leu Gly Gly Ala Asn  
               
               
                 215                 220                 225                 230  
               
               
                   
               
               
                 gag ggc acg cgt acg gcg ctt tca gtg gtc act tgg aac cag aaa ggg      896  
               
               
                 Glu Gly Thr Arg Thr Ala Leu Ser Val Val Thr Trp Asn Gln Lys Gly  
               
               
                                 235                 240                 245  
               
               
                   
               
               
                 gtg acc att agg gat acc ccc gaa ggt tct gaa ccg tgg tca cta gtt      944  
               
               
                 Val Thr Ile Arg Asp Thr Pro Glu Gly Ser Glu Pro Trp Ser Leu Val  
               
               
                             250                 255                 260  
               
               
                   
               
               
                 aca gcg cta tgc gtg ctt tcg aat gtc acg ttc cca tgc gac aaa cca      992  
               
               
                 Thr Ala Leu Cys Val Leu Ser Asn Val Thr Phe Pro Cys Asp Lys Pro  
               
               
                         265                 270                 275  
               
               
                   
               
               
                 ccc gtg tgc tat tca ctg acg cca gaa cga aca ctc gac gtg ctc gaa     1040  
               
               
                 Pro Val Cys Tyr Ser Leu Thr Pro Glu Arg Thr Leu Asp Val Leu Glu  
               
               
                     280                 285                 290  
               
               
                   
               
               
                 gag aac gtc gac aat cca aat tac gac acg ctg ctg gag aac gtc ttg     1088  
               
               
                 Glu Asn Val Asp Asn Pro Asn Tyr Asp Thr Leu Leu Glu Asn Val Leu  
               
               
                 295                 300                 305                 310  
               
               
                   
               
               
                 aaa tgt cca tca cgc cgg ccc aaa cga agc att acc gat gac ttc aca     1136  
               
               
                 Lys Cys Pro Ser Arg Arg Pro Lys Arg Ser Ile Thr Asp Asp Phe Thr  
               
               
                                 315                 320                 325  
               
               
                   
               
               
                 ctg acc agt ccc tac ctg ggg ttc tgc ccg tat tgc aga cac tca acg     1184  
               
               
                 Leu Thr Ser Pro Tyr Leu Gly Phe Cys Pro Tyr Cys Arg His Ser Thr  
               
               
                             330                 335                 340  
               
               
                   
               
               
                 ccg tgt ttc agc cca ata aaa att gag aac gtg tgg gac gaa tct gat     1232  
               
               
                 Pro Cys Phe Ser Pro Ile Lys Ile Glu Asn Val Trp Asp Glu Ser Asp  
               
               
                         345                 350                 355  
               
               
                   
               
               
                 gat gga tcg att aga atc cag gtc tcg gca caa ttc ggc tac aat cag     1280  
               
               
                 Asp Gly Ser Ile Arg Ile Gln Val Ser Ala Gln Phe Gly Tyr Asn Gln  
               
               
                     360                 365                 370  
               
               
                   
               
               
                 gca ggc act gcg gat gtc acc aaa ttc cgt tac atg tct ttc gac cac     1328  
               
               
                 Ala Gly Thr Ala Asp Val Thr Lys Phe Arg Tyr Met Ser Phe Asp His  
               
               
                 375                 380                 385                 390  
               
               
                   
               
               
                 gac cat gac atc aag gaa gac agt atg gag aaa ata gct atc agc aca     1376  
               
               
                 Asp His Asp Ile Lys Glu Asp Ser Met Glu Lys Ile Ala Ile Ser Thr  
               
               
                                 395                 400                 405  
               
               
                   
               
               
                 tct gga ccc tgc cgt cgt ctt ggc cac aaa ggg tac ttc ctg tta gct     1424  
               
               
                 Ser Gly Pro Cys Arg Arg Leu Gly His Lys Gly Tyr Phe Leu Leu Ala  
               
               
                             410                 415                 420  
               
               
                   
               
               
                 caa tgt cct cca ggt gac agt gta acc gtc agt atc acg agc gga gca     1472  
               
               
                 Gln Cys Pro Pro Gly Asp Ser Val Thr Val Ser Ile Thr Ser Gly Ala  
               
               
                         425                 430                 435  
               
               
                   
               
               
                 tct gag aat tca tgc acc gtg gag aaa aag atc agg agg aag ttt gtc     1520  
               
               
                 Ser Glu Asn Ser Cys Thr Val Glu Lys Lys Ile Arg Arg Lys Phe Val  
               
               
                     440                 445                 450  
               
               
                   
               
               
                 ggt aga gag gag tac ttg ttc cca ccc gtc cat gga aag ctg gta aag     1568  
               
               
                 Gly Arg Glu Glu Tyr Leu Phe Pro Pro Val His Gly Lys Leu Val Lys  
               
               
                 455                 460                 465                 470  
               
               
                   
               
               
                 tgc cac gtt tac gat cac ttg aag gag acg tct gcc ggg tac ata acc     1616  
               
               
                 Cys His Val Tyr Asp His Leu Lys Glu Thr Ser Ala Gly Tyr Ile Thr  
               
               
                                 475                 480                 485  
               
               
                   
               
               
                 atg cac agg cca ggc cca cac gcg tat aag tcc tat ctg gag gaa gcg     1664  
               
               
                 Met His Arg Pro Gly Pro His Ala Tyr Lys Ser Tyr Leu Glu Glu Ala  
               
               
                             490                 495                 500  
               
               
                   
               
               
                 tca ggc gaa gtg tac att aaa cca cct tct ggc aag aac gtc acc tac     1712  
               
               
                 Ser Gly Glu Val Tyr Ile Lys Pro Pro Ser Gly Lys Asn Val Thr Tyr  
               
               
                         505                 510                 515  
               
               
                   
               
               
                 gaa tgt aag tgt ggc gac tac agc aca ggt atc gtg agc acg cga acg     1760  
               
               
                 Glu Cys Lys Cys Gly Asp Tyr Ser Thr Gly Ile Val Ser Thr Arg Thr  
               
               
                     520                 525                 530  
               
               
                   
               
               
                 aag atg aac ggc tgc act aaa gca aaa cag tgc att gcc tac aag agc     1808  
               
               
                 Lys Met Asn Gly Cys Thr Lys Ala Lys Gln Cys Ile Ala Tyr Lys Ser  
               
               
                 535                 540                 545                 550  
               
               
                   
               
               
                 gac caa acg aaa tgg gtc ttc aac tcg ccg gat ctt att agg cac aca     1856  
               
               
                 Asp Gln Thr Lys Trp Val Phe Asn Ser Pro Asp Leu Ile Arg His Thr  
               
               
                                 555                 560                 565  
               
               
                   
               
               
                 gac cac tca gtg caa ggt aaa ttg cac att cca ttc cgc ttg aca ccg     1904  
               
               
                 Asp His Ser Val Gln Gly Lys Leu His Ile Pro Phe Arg Leu Thr Pro  
               
               
                             570                 575                 580  
               
               
                   
               
               
                 aca gtc tgc ccg gtt ccg tta gct cac acg cct aca gtc acg aag tgg     1952  
               
               
                 Thr Val Cys Pro Val Pro Leu Ala His Thr Pro Thr Val Thr Lys Trp  
               
               
                         585                 590                 595  
               
               
                   
               
               
                 ttc aaa ggc atc acc ctc cac ctg act gca atg cga cca aca ttg ctg     2000  
               
               
                 Phe Lys Gly Ile Thr Leu His Leu Thr Ala Met Arg Pro Thr Leu Leu  
               
               
                     600                 605                 610  
               
               
                   
               
               
                 aca acg aga aaa ttg ggg ctg cga gca gac gca aca gca gaa tgg att     2048  
               
               
                 Thr Thr Arg Lys Leu Gly Leu Arg Ala Asp Ala Thr Ala Glu Trp Ile  
               
               
                 615                 620                 625                 630  
               
               
                   
               
               
                 aca ggg tct aca tcc agg aat ttt tct gtg ggg cga gaa ggg ctg gag     2096  
               
               
                 Thr Gly Ser Thr Ser Arg Asn Phe Ser Val Gly Arg Glu Gly Leu Glu  
               
               
                                 635                 640                 645  
               
               
                   
               
               
                 tac gta tgg ggt aac cat gaa cca gtc aga gtc tgg gcc cag gag tcg     2144  
               
               
                 Tyr Val Trp Gly Asn His Glu Pro Val Arg Val Trp Ala Gln Glu Ser  
               
               
                             650                 655                 660  
               
               
                   
               
               
                 gca cca ggc gac cca cat gga tgg ccg cat gag atc atc atc cac tat     2192  
               
               
                 Ala Pro Gly Asp Pro His Gly Trp Pro His Glu Ile Ile Ile His Tyr  
               
               
                         665                 670                 675  
               
               
                   
               
               
                 tat cat cgg cat cca gtc tac act gtc att gtg ctg tgt ggt gtc gct     2240  
               
               
                 Tyr His Arg His Pro Val Tyr Thr Val Ile Val Leu Cys Gly Val Ala  
               
               
                     680                 685                 690  
               
               
                   
               
               
                 ctt gct atc ctg gta ggc act gca tca tca gca gct tgc atc gcc aaa     2288  
               
               
                 Leu Ala Ile Leu Val Gly Thr Ala Ser Ser Ala Ala Cys Ile Ala Lys  
               
               
                 695                 700                 705                 710  
               
               
                   
               
               
                 gca aga aga gac tgc ctg acg cca tac gcg ctt gca ccg aac gca acg     2336  
               
               
                 Ala Arg Arg Asp Cys Leu Thr Pro Tyr Ala Leu Ala Pro Asn Ala Thr  
               
               
                                 715                 720                 725  
               
               
                   
               
               
                 gta ccc aca gca tta gcg gtt ttg tgc tgc att cgg cca acc aac gct     2384  
               
               
                 Val Pro Thr Ala Leu Ala Val Leu Cys Cys Ile Arg Pro Thr Asn Ala  
               
               
                             730                 735                 740  
               
               
                   
               
               
                 gaa aca ttt gga gaa act ttg aac cat ctg tgg ttt aac aac caa ccg     2432  
               
               
                 Glu Thr Phe Gly Glu Thr Leu Asn His Leu Trp Phe Asn Asn Gln Pro  
               
               
                         745                 750                 755  
               
               
                   
               
               
                 ttt ctc tgg gca cag ttg tgc att cct ctg gca gcg ctt gtt att ctg     2480  
               
               
                 Phe Leu Trp Ala Gln Leu Cys Ile Pro Leu Ala Ala Leu Val Ile Leu  
               
               
                     760                 765                 770  
               
               
                   
               
               
                 ttc cgc tgc ttt tca tgc tgc atg cct ttt tta ttg gtt gca ggc gtc     2528  
               
               
                 Phe Arg Cys Phe Ser Cys Cys Met Pro Phe Leu Leu Val Ala Gly Val  
               
               
                 775                 780                 785                 790  
               
               
                   
               
               
                 tgc ctg ggg aag gta gac gcc ttc gaa cat gcg acc act gtg cca aat     2576  
               
               
                 Cys Leu Gly Lys Val Asp Ala Phe Glu His Ala Thr Thr Val Pro Asn  
               
               
                                 795                 800                 805  
               
               
                   
               
               
                 gtt ccg ggg atc ccg tat aag gcg ttg gtc gaa cgc gca ggt tac gcg     2624  
               
               
                 Val Pro Gly Ile Pro Tyr Lys Ala Leu Val Glu Arg Ala Gly Tyr Ala  
               
               
                             810                 815                 820  
               
               
                   
               
               
                 cca ctt aac ctg gag atc acg gtc gtc tca tcg gaa tta aca cct tca     2672  
               
               
                 Pro Leu Asn Leu Glu Ile Thr Val Val Ser Ser Glu Leu Thr Pro Ser  
               
               
                         825                 830                 835  
               
               
                   
               
               
                 act aac aag gag tac gtg acc tgc aaa ttc cac aca gtc att cct tca     2720  
               
               
                 Thr Asn Lys Glu Tyr Val Thr Cys Lys Phe His Thr Val Ile Pro Ser  
               
               
                     840                 845                 850  
               
               
                   
               
               
                 cca caa gtt aaa tgc tgc ggg tcc ctc gag tgc aag gca tcc tca aag     2768  
               
               
                 Pro Gln Val Lys Cys Cys Gly Ser Leu Glu Cys Lys Ala Ser Ser Lys  
               
               
                 855                 860                 865                 870  
               
               
                   
               
               
                 gcg gat tac aca tgc cgc gtt ttt ggc ggt gtg tac cct ttc atg tgg     2816  
               
               
                 Ala Asp Tyr Thr Cys Arg Val Phe Gly Gly Val Tyr Pro Phe Met Trp  
               
               
                                 875                 880                 885  
               
               
                   
               
               
                 gga ggc gca caa tgc ttc tgt gac agt gag aac aca caa ctg agt gag     2864  
               
               
                 Gly Gly Ala Gln Cys Phe Cys Asp Ser Glu Asn Thr Gln Leu Ser Glu  
               
               
                             890                 895                 900  
               
               
                   
               
               
                 gcg tac gtc gag ttc gct cca gac tgc act ata gat cac gca gtc gca     2912  
               
               
                 Ala Tyr Val Glu Phe Ala Pro Asp Cys Thr Ile Asp His Ala Val Ala  
               
               
                         905                 910                 915  
               
               
                   
               
               
                 cta aaa gtt cac aca gct gct ctg aaa gtc ggc ctg cgt ata gta tac     2960  
               
               
                 Leu Lys Val His Thr Ala Ala Leu Lys Val Gly Leu Arg Ile Val Tyr  
               
               
                     920                 925                 930  
               
               
                   
               
               
                 ggc aac acc acc gcg cac ctg gat acg ttt gtc aat ggc gtc acg cca     3008  
               
               
                 Gly Asn Thr Thr Ala His Leu Asp Thr Phe Val Asn Gly Val Thr Pro  
               
               
                 935                 940                 945                 950  
               
               
                   
               
               
                 ggt tcc tca cgg gac ctg aag gtc ata gca ggg ccg ata tca gcc gct     3056  
               
               
                 Gly Ser Ser Arg Asp Leu Lys Val Ile Ala Gly Pro Ile Ser Ala Ala  
               
               
                                 955                 960                 965  
               
               
                   
               
               
                 ttt tca ccc ttt gac cat aag gtc gtc atc aga aag ggg ctt gtt tac     3104  
               
               
                 Phe Ser Pro Phe Asp His Lys Val Val Ile Arg Lys Gly Leu Val Tyr  
               
               
                             970                 975                 980  
               
               
                   
               
               
                 aac tac gac ttc cct gag tat gga gct atg aaa cca gga gcg ttc ggc     3152  
               
               
                 Asn Tyr Asp Phe Pro Glu Tyr Gly Ala Met Lys Pro Gly Ala Phe Gly  
               
               
                         985                 990                 995  
               
               
                   
               
               
                 gat att  caa gca tcc tcg ctt  gat gct aca gac ata  gta gcc cgc      3197  
               
               
                 Asp Ile  Gln Ala Ser Ser Leu  Asp Ala Thr Asp Ile  Val Ala Arg  
               
               
                     1000                 1005                 1010  
               
               
                   
               
               
                 act gac  ata cgg ctg ctg aag  cct tct gtc aag aac  atc cac gtc      3242  
               
               
                 Thr Asp  Ile Arg Leu Leu Lys  Pro Ser Val Lys Asn  Ile His Val  
               
               
                     1015                 1020                 1025  
               
               
                   
               
               
                 ccc tac  acc caa gca gta tca  ggg tat gaa atg tgg  aag aac aac      3287  
               
               
                 Pro Tyr  Thr Gln Ala Val Ser  Gly Tyr Glu Met Trp  Lys Asn Asn  
               
               
                     1030                 1035                 1040  
               
               
                   
               
               
                 tca gga  cga ccc ctg caa gaa  aca gca cca ttt gga  tgt aaa att      3332  
               
               
                 Ser Gly  Arg Pro Leu Gln Glu  Thr Ala Pro Phe Gly  Cys Lys Ile  
               
               
                     1045                 1050                 1055  
               
               
                   
               
               
                 gaa gtg  gag cct ctg cga gcg  tct aac tgt gct tac  ggg cac atc      3377  
               
               
                 Glu Val  Glu Pro Leu Arg Ala  Ser Asn Cys Ala Tyr  Gly His Ile  
               
               
                     1060                 1065                 1070  
               
               
                   
               
               
                 cct atc  tcg att gac atc cct  gat gca gct ttt gtg  aga tca tca      3422  
               
               
                 Pro Ile  Ser Ile Asp Ile Pro  Asp Ala Ala Phe Val  Arg Ser Ser  
               
               
                     1075                 1080                 1085  
               
               
                   
               
               
                 gaa tca  cca aca att tta gaa  gtt agc tgc aca gta  gca gac tgc      3467  
               
               
                 Glu Ser  Pro Thr Ile Leu Glu  Val Ser Cys Thr Val  Ala Asp Cys  
               
               
                     1090                 1095                 1100  
               
               
                   
               
               
                 att tat  tct gca gac ttt ggt  ggt tct cta aca tta  cag tac aaa      3512  
               
               
                 Ile Tyr  Ser Ala Asp Phe Gly  Gly Ser Leu Thr Leu  Gln Tyr Lys  
               
               
                     1105                 1110                 1115  
               
               
                   
               
               
                 gct gac  agg gag gga cat tgt  cca gtt cac tcc cac  tcc acg aca      3557  
               
               
                 Ala Asp  Arg Glu Gly His Cys  Pro Val His Ser His  Ser Thr Thr  
               
               
                     1120                 1125                 1130  
               
               
                   
               
               
                 gct gtt  ttg aag gaa gcg acc  aca cat gtg act gcc  gta ggc agc      3602  
               
               
                 Ala Val  Leu Lys Glu Ala Thr  Thr His Val Thr Ala  Val Gly Ser  
               
               
                     1135                 1140                 1145  
               
               
                   
               
               
                 ata aca  cta cat ttt agc aca  tcg agc cca caa gca  aat ttt ata      3647  
               
               
                 Ile Thr  Leu His Phe Ser Thr  Ser Ser Pro Gln Ala  Asn Phe Ile  
               
               
                     1150                 1155                 1160  
               
               
                   
               
               
                 gtt tcg  cta tgc ggc aag aag  tcc acc tgc aat gct  gaa tgt aaa      3692  
               
               
                 Val Ser  Leu Cys Gly Lys Lys  Ser Thr Cys Asn Ala  Glu Cys Lys  
               
               
                     1165                 1170                 1175  
               
               
                   
               
               
                 cca ccg  gcc gac cac ata att  gga gaa cca cat aaa  gtc gac caa      3737  
               
               
                 Pro Pro  Ala Asp His Ile Ile  Gly Glu Pro His Lys  Val Asp Gln  
               
               
                     1180                 1185                 1190  
               
               
                   
               
               
                 gaa ttc  cag gcg gca gtt tcc  aaa aca tct tgg aac  tgg ctg ctt      3782  
               
               
                 Glu Phe  Gln Ala Ala Val Ser  Lys Thr Ser Trp Asn  Trp Leu Leu  
               
               
                     1195                 1200                 1205  
               
               
                   
               
               
                 gca ctg  ttt ggg gga gca tca  tcc ctc att gtt gta  gga ctt ata      3827  
               
               
                 Ala Leu  Phe Gly Gly Ala Ser  Ser Leu Ile Val Val  Gly Leu Ile  
               
               
                     1210                 1215                 1220  
               
               
                   
               
               
                 gtg ttg  gtc tgc agc tct atg  ctt ata aac aca cgt  aga tga          3869  
               
               
                 Val Leu  Val Cys Ser Ser Met  Leu Ile Asn Thr Arg  Arg  
               
               
                     1225                 1230                 1235  
               
               
                   
               
               
                 ctgagcgcgg acactgacat agcggtaaaa ctcgatgtac ttccgaggaa gcgtggtgca   3929  
               
               
                   
               
               
                 taatgccacg cgccgcttga cactaaaact cgatgtattt ccgaggaagc acagtgcata   3989  
               
               
                   
               
               
                 atgctgtgca gtgtcacatt aatcgtatat cacactacat attaacaaca ctatatcact   4049  
               
               
                   
               
               
                 tttatgagac tcactatggg tctctaatat acactacaca tattttactt aaaaacacta   4109  
               
               
                   
               
               
                 tacacacttt ataaattctc tcataatttc actttaggtt t                       4150  
               
               
                   
               
               
                   
               
               
                 &lt;210&gt; SEQ ID NO 6  
               
               
                 &lt;211&gt; LENGTH: 1236  
               
               
                 &lt;212&gt; TYPE: PRT  
               
               
                 &lt;213&gt; ORGANISM: Western equine encephalomyelitis virus - strain 71V-1658  
               
               
                   
               
               
                 &lt;400&gt; SEQUENCE: 6  
               
               
                   
               
               
                 Met Phe Pro Tyr Pro Gln Leu Asn Phe Pro Pro Val Tyr Pro Thr Asn  
               
               
                 1               5                   10                  15  
               
               
                   
               
               
                 Pro Met Ala Tyr Arg Asp Pro Asn Pro Pro Arg Arg Arg Trp Arg Pro  
               
               
                             20                  25                  30  
               
               
                   
               
               
                 Phe Arg Pro Pro Leu Ala Ala Gln Ile Glu Asp Leu Arg Arg Ser Ile  
               
               
                         35                  40                  45  
               
               
                   
               
               
                 Val Asn Leu Thr Phe Lys Gln Arg Ser Pro Asn Pro Pro Pro Gly Pro  
               
               
                     50                  55                  60  
               
               
                   
               
               
                 Pro Pro Lys Lys Lys Lys Ser Ala Pro Lys Pro Lys Pro Thr Gln Pro  
               
               
                 65                  70                  75                  80  
               
               
                   
               
               
                 Lys Lys Lys Lys Gln Gln Ala Lys Arg Thr Lys Arg Lys Pro Lys Pro  
               
               
                                 85                  90                  95  
               
               
                   
               
               
                 Gly Lys Arg Gln Arg Met Cys Met Lys Leu Glu Ser Asp Lys Thr Phe  
               
               
                             100                 105                 110  
               
               
                   
               
               
                 Pro Ile Met Leu Asn Gly Gln Val Asn Gly Tyr Ala Cys Val Val Gly  
               
               
                         115                 120                 125  
               
               
                   
               
               
                 Gly Arg Leu Met Lys Pro Leu His Val Glu Gly Lys Ile Asp Asn Glu  
               
               
                     130                 135                 140  
               
               
                   
               
               
                 Gln Leu Ala Ala Val Lys Leu Lys Lys Ala Ser Met Tyr Asp Leu Glu  
               
               
                 145                 150                 155                 160  
               
               
                   
               
               
                 Tyr Gly Asp Val Pro Gln Asn Met Lys Ser Asp Thr Leu Gln Tyr Thr  
               
               
                                 165                 170                 175  
               
               
                   
               
               
                 Ser Asp Lys Pro Pro Gly Phe Tyr Asn Trp His His Gly Ala Val Gln  
               
               
                             180                 185                 190  
               
               
                   
               
               
                 Tyr Glu Asn Gly Arg Phe Thr Val Pro Arg Gly Val Gly Gly Lys Gly  
               
               
                         195                 200                 205  
               
               
                   
               
               
                 Asp Ser Gly Arg Pro Ile Leu Asp Asn Arg Gly Arg Val Val Ala Ile  
               
               
                     210                 215                 220  
               
               
                   
               
               
                 Val Leu Gly Gly Ala Asn Glu Gly Thr Arg Thr Ala Leu Ser Val Val  
               
               
                 225                 230                 235                 240  
               
               
                   
               
               
                 Thr Trp Asn Gln Lys Gly Val Thr Ile Arg Asp Thr Pro Glu Gly Ser  
               
               
                                 245                 250                 255  
               
               
                   
               
               
                 Glu Pro Trp Ser Leu Val Thr Ala Leu Cys Val Leu Ser Asn Val Thr  
               
               
                             260                 265                 270  
               
               
                   
               
               
                 Phe Pro Cys Asp Lys Pro Pro Val Cys Tyr Ser Leu Thr Pro Glu Arg  
               
               
                         275                 280                 285  
               
               
                   
               
               
                 Thr Leu Asp Val Leu Glu Glu Asn Val Asp Asn Pro Asn Tyr Asp Thr  
               
               
                     290                 295                 300  
               
               
                   
               
               
                 Leu Leu Glu Asn Val Leu Lys Cys Pro Ser Arg Arg Pro Lys Arg Ser  
               
               
                 305                 310                 315                 320  
               
               
                   
               
               
                 Ile Thr Asp Asp Phe Thr Leu Thr Ser Pro Tyr Leu Gly Phe Cys Pro  
               
               
                                 325                 330                 335  
               
               
                   
               
               
                 Tyr Cys Arg His Ser Thr Pro Cys Phe Ser Pro Ile Lys Ile Glu Asn  
               
               
                             340                 345                 350  
               
               
                   
               
               
                 Val Trp Asp Glu Ser Asp Asp Gly Ser Ile Arg Ile Gln Val Ser Ala  
               
               
                         355                 360                 365  
               
               
                   
               
               
                 Gln Phe Gly Tyr Asn Gln Ala Gly Thr Ala Asp Val Thr Lys Phe Arg  
               
               
                     370                 375                 380  
               
               
                   
               
               
                 Tyr Met Ser Phe Asp His Asp His Asp Ile Lys Glu Asp Ser Met Glu  
               
               
                 385                 390                 395                 400  
               
               
                   
               
               
                 Lys Ile Ala Ile Ser Thr Ser Gly Pro Cys Arg Arg Leu Gly His Lys  
               
               
                                 405                 410                 415  
               
               
                   
               
               
                 Gly Tyr Phe Leu Leu Ala Gln Cys Pro Pro Gly Asp Ser Val Thr Val  
               
               
                             420                 425                 430  
               
               
                   
               
               
                 Ser Ile Thr Ser Gly Ala Ser Glu Asn Ser Cys Thr Val Glu Lys Lys  
               
               
                         435                 440                 445  
               
               
                   
               
               
                 Ile Arg Arg Lys Phe Val Gly Arg Glu Glu Tyr Leu Phe Pro Pro Val  
               
               
                     450                 455                 460  
               
               
                   
               
               
                 His Gly Lys Leu Val Lys Cys His Val Tyr Asp His Leu Lys Glu Thr  
               
               
                 465                 470                 475                 480  
               
               
                   
               
               
                 Ser Ala Gly Tyr Ile Thr Met His Arg Pro Gly Pro His Ala Tyr Lys  
               
               
                                 485                 490                 495  
               
               
                   
               
               
                 Ser Tyr Leu Glu Glu Ala Ser Gly Glu Val Tyr Ile Lys Pro Pro Ser  
               
               
                             500                 505                 510  
               
               
                   
               
               
                 Gly Lys Asn Val Thr Tyr Glu Cys Lys Cys Gly Asp Tyr Ser Thr Gly  
               
               
                         515                 520                 525  
               
               
                   
               
               
                 Ile Val Ser Thr Arg Thr Lys Met Asn Gly Cys Thr Lys Ala Lys Gln  
               
               
                     530                 535                 540  
               
               
                   
               
               
                 Cys Ile Ala Tyr Lys Ser Asp Gln Thr Lys Trp Val Phe Asn Ser Pro  
               
               
                 545                 550                 555                 560  
               
               
                   
               
               
                 Asp Leu Ile Arg His Thr Asp His Ser Val Gln Gly Lys Leu His Ile  
               
               
                                 565                 570                 575  
               
               
                   
               
               
                 Pro Phe Arg Leu Thr Pro Thr Val Cys Pro Val Pro Leu Ala His Thr  
               
               
                             580                 585                 590  
               
               
                   
               
               
                 Pro Thr Val Thr Lys Trp Phe Lys Gly Ile Thr Leu His Leu Thr Ala  
               
               
                         595                 600                 605  
               
               
                   
               
               
                 Met Arg Pro Thr Leu Leu Thr Thr Arg Lys Leu Gly Leu Arg Ala Asp  
               
               
                     610                 615                 620  
               
               
                   
               
               
                 Ala Thr Ala Glu Trp Ile Thr Gly Ser Thr Ser Arg Asn Phe Ser Val  
               
               
                 625                 630                 635                 640  
               
               
                   
               
               
                 Gly Arg Glu Gly Leu Glu Tyr Val Trp Gly Asn His Glu Pro Val Arg  
               
               
                                 645                 650                 655  
               
               
                   
               
               
                 Val Trp Ala Gln Glu Ser Ala Pro Gly Asp Pro His Gly Trp Pro His  
               
               
                             660                 665                 670  
               
               
                   
               
               
                 Glu Ile Ile Ile His Tyr Tyr His Arg His Pro Val Tyr Thr Val Ile  
               
               
                         675                 680                 685  
               
               
                   
               
               
                 Val Leu Cys Gly Val Ala Leu Ala Ile Leu Val Gly Thr Ala Ser Ser  
               
               
                     690                 695                 700  
               
               
                   
               
               
                 Ala Ala Cys Ile Ala Lys Ala Arg Arg Asp Cys Leu Thr Pro Tyr Ala  
               
               
                 705                 710                 715                 720  
               
               
                   
               
               
                 Leu Ala Pro Asn Ala Thr Val Pro Thr Ala Leu Ala Val Leu Cys Cys  
               
               
                                 725                 730                 735  
               
               
                   
               
               
                 Ile Arg Pro Thr Asn Ala Glu Thr Phe Gly Glu Thr Leu Asn His Leu  
               
               
                             740                 745                 750  
               
               
                   
               
               
                 Trp Phe Asn Asn Gln Pro Phe Leu Trp Ala Gln Leu Cys Ile Pro Leu  
               
               
                         755                 760                 765  
               
               
                   
               
               
                 Ala Ala Leu Val Ile Leu Phe Arg Cys Phe Ser Cys Cys Met Pro Phe  
               
               
                     770                 775                 780  
               
               
                   
               
               
                 Leu Leu Val Ala Gly Val Cys Leu Gly Lys Val Asp Ala Phe Glu His  
               
               
                 785                 790                 795                 800  
               
               
                   
               
               
                 Ala Thr Thr Val Pro Asn Val Pro Gly Ile Pro Tyr Lys Ala Leu Val  
               
               
                                 805                 810                 815  
               
               
                   
               
               
                 Glu Arg Ala Gly Tyr Ala Pro Leu Asn Leu Glu Ile Thr Val Val Ser  
               
               
                             820                 825                 830  
               
               
                   
               
               
                 Ser Glu Leu Thr Pro Ser Thr Asn Lys Glu Tyr Val Thr Cys Lys Phe  
               
               
                         835                 840                 845  
               
               
                   
               
               
                 His Thr Val Ile Pro Ser Pro Gln Val Lys Cys Cys Gly Ser Leu Glu  
               
               
                     850                 855                 860  
               
               
                   
               
               
                 Cys Lys Ala Ser Ser Lys Ala Asp Tyr Thr Cys Arg Val Phe Gly Gly  
               
               
                 865                 870                 875                 880  
               
               
                   
               
               
                 Val Tyr Pro Phe Met Trp Gly Gly Ala Gln Cys Phe Cys Asp Ser Glu  
               
               
                                 885                 890                 895  
               
               
                   
               
               
                 Asn Thr Gln Leu Ser Glu Ala Tyr Val Glu Phe Ala Pro Asp Cys Thr  
               
               
                             900                 905                 910  
               
               
                   
               
               
                 Ile Asp His Ala Val Ala Leu Lys Val His Thr Ala Ala Leu Lys Val  
               
               
                         915                 920                 925  
               
               
                   
               
               
                 Gly Leu Arg Ile Val Tyr Gly Asn Thr Thr Ala His Leu Asp Thr Phe  
               
               
                     930                 935                 940  
               
               
                   
               
               
                 Val Asn Gly Val Thr Pro Gly Ser Ser Arg Asp Leu Lys Val Ile Ala  
               
               
                 945                 950                 955                 960  
               
               
                   
               
               
                 Gly Pro Ile Ser Ala Ala Phe Ser Pro Phe Asp His Lys Val Val Ile  
               
               
                                 965                 970                 975  
               
               
                   
               
               
                 Arg Lys Gly Leu Val Tyr Asn Tyr Asp Phe Pro Glu Tyr Gly Ala Met  
               
               
                             980                 985                 990  
               
               
                   
               
               
                 Lys Pro Gly Ala Phe Gly Asp Ile  Gln Ala Ser Ser Leu  Asp Ala Thr  
               
               
                         995                 1000                 1005  
               
               
                   
               
               
                 Asp Ile  Val Ala Arg Thr Asp  Ile Arg Leu Leu Lys  Pro Ser Val  
               
               
                     1010                 1015                 1020  
               
               
                   
               
               
                 Lys Asn  Ile His Val Pro Tyr  Thr Gln Ala Val Ser  Gly Tyr Glu  
               
               
                     1025                 1030                 1035  
               
               
                   
               
               
                 Met Trp  Lys Asn Asn Ser Gly  Arg Pro Leu Gln Glu  Thr Ala Pro  
               
               
                     1040                 1045                 1050  
               
               
                   
               
               
                 Phe Gly  Cys Lys Ile Glu Val  Glu Pro Leu Arg Ala  Ser Asn Cys  
               
               
                     1055                 1060                 1065  
               
               
                   
               
               
                 Ala Tyr  Gly His Ile Pro Ile  Ser Ile Asp Ile Pro  Asp Ala Ala  
               
               
                     1070                 1075                 1080  
               
               
                   
               
               
                 Phe Val  Arg Ser Ser Glu Ser  Pro Thr Ile Leu Glu  Val Ser Cys  
               
               
                     1085                 1090                 1095  
               
               
                   
               
               
                 Thr Val  Ala Asp Cys Ile Tyr  Ser Ala Asp Phe Gly  Gly Ser Leu  
               
               
                     1100                 1105                 1110  
               
               
                   
               
               
                 Thr Leu  Gln Tyr Lys Ala Asp  Arg Glu Gly His Cys  Pro Val His  
               
               
                     1115                 1120                 1125  
               
               
                   
               
               
                 Ser His  Ser Thr Thr Ala Val  Leu Lys Glu Ala Thr  Thr His Val  
               
               
                     1130                 1135                 1140  
               
               
                   
               
               
                 Thr Ala  Val Gly Ser Ile Thr  Leu His Phe Ser Thr  Ser Ser Pro  
               
               
                     1145                 1150                 1155  
               
               
                   
               
               
                 Gln Ala  Asn Phe Ile Val Ser  Leu Cys Gly Lys Lys  Ser Thr Cys  
               
               
                     1160                 1165                 1170  
               
               
                   
               
               
                 Asn Ala  Glu Cys Lys Pro Pro  Ala Asp His Ile Ile  Gly Glu Pro  
               
               
                     1175                 1180                 1185  
               
               
                   
               
               
                 His Lys  Val Asp Gln Glu Phe  Gln Ala Ala Val Ser  Lys Thr Ser  
               
               
                     1190                 1195                 1200  
               
               
                   
               
               
                 Trp Asn  Trp Leu Leu Ala Leu  Phe Gly Gly Ala Ser  Ser Leu Ile  
               
               
                     1205                 1210                 1215  
               
               
                   
               
               
                 Val Val  Gly Leu Ile Val Leu  Val Cys Ser Ser Met  Leu Ile Asn  
               
               
                     1220                 1225                 1230  
               
               
                   
               
               
                 Thr Arg  Arg  
               
               
                     1235  
               
               
                   
               
               
                   
               
               
                 &lt;210&gt; SEQ ID NO 7  
               
               
                 &lt;211&gt; LENGTH: 4395  
               
               
                 &lt;212&gt; TYPE: DNA  
               
               
                 &lt;213&gt; ORGANISM: Western equine encephalomyelitis virus - STRAIN 71v-1658  
               
               
                 &lt;220&gt; FEATURE:  
               
               
                 &lt;221&gt; NAME/KEY: CMV promoter  
               
               
                 &lt;222&gt; LOCATION: (1)..(1260)  
               
               
                 &lt;223&gt; OTHER INFORMATION: Pvax vector sequence: 1-196; CMV promoter:  
               
               
                       1-115; CMV putative transcriptional start site: 125; T7 promoter:  
               
               
                       48-167; pVAX multicloning region: 168-196; polyprotein (C-E3-E2-  
               
               
                       6K-E1): 214-4065; pcDW-HX45 nontranslated region: 4066-4348; pcDW-  
               
               
                       HX45 vector sequence:4349-4385; pVAX vector sequence: 4386  
               
               
                   
               
               
                 &lt;400&gt; SEQUENCE: 7  
               
               
                   
               
               
                 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg     60  
               
               
                   
               
               
                 gcggtaggcg tgtacggtgg gaggtcatat ataagcagag tctctctggc taactagaga    120  
               
               
                   
               
               
                 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg    180  
               
               
                   
               
               
                 ctagcgttta aacttaagct tggtaccgag ctcatactgg caggcctgat catcacgtct    240  
               
               
                   
               
               
                 ctgtccacgt tagccgaaag cgttaagaac ttcaagagca taagagggag cccaatcacc    300  
               
               
                   
               
               
                 ctctacggct gacctaaata ggtgacgtag tagacacgca cctacccacc gccagaatgt    360  
               
               
                   
               
               
                 ttccataccc tcagctgaac tttccaccag tttaccctac aaatccgatg gcttaccgag    420  
               
               
                   
               
               
                 atccaaaccc tcctaggcgc cgctggaggc cgtttcggcc cccgctggct gctcaaatcg    480  
               
               
                   
               
               
                 aagatcttag gaggtcgata gtcaacttga ctttcaaaca acgatcacct aatccgccgc    540  
               
               
                   
               
               
                 caggtccacc gccaaagaag aagaagagtg ctcctaagcc aaaacctact cagcctaaaa    600  
               
               
                   
               
               
                 agaagaagca gcaagccaag aggacgaaac gcaagcctaa accagggaaa cgacaacgta    660  
               
               
                   
               
               
                 tgtgtatgaa gttggagtcg gacaagacat ttccgatcat gctgaacggc caagtgaatg    720  
               
               
                   
               
               
                 gatatgcctg cgttgtcgga ggaaggctga tgaaaccact ccacgttgaa ggaaaaattg    780  
               
               
                   
               
               
                 ataatgagca attagcggcc gtgaaattga agaaggctag catgtacgac ttggagtacg    840  
               
               
                   
               
               
                 gcgacgttcc ccagaacatg aaatcagaca cgctgcagta caccagcgac aaaccaccgg    900  
               
               
                   
               
               
                 gcttctacaa ctggcaccac ggcgcagtcc agtatgagaa tgggagattt accgtaccga    960  
               
               
                   
               
               
                 gaggagtggg cgggaaaggc gacagcggaa gaccgatcct ggacaacaga ggcagagttg   1020  
               
               
                   
               
               
                 tggctattgt tctaggaggt gcaaatgagg gcacgcgtac ggcgctttca gtggtcactt   1080  
               
               
                   
               
               
                 ggaaccagaa aggggtgacc attagggata cccccgaagg ttctgaaccg tggtcactag   1140  
               
               
                   
               
               
                 ttacagcgct atgcgtgctt tcgaatgtca cgttcccatg cgacaaacca cccgtgtgct   1200  
               
               
                   
               
               
                 attcactgac gccagaacga acactcgacg tgctcgaaga gaacgtcgac aatccaaatt   1260  
               
               
                   
               
               
                 acgacacgct gctggagaac gtcttgaaat gtccatcacg ccggcccaaa cgaagcatta   1320  
               
               
                   
               
               
                 ccgatgactt cacactgacc agtccctacc tggggttctg cccgtattgc agacactcaa   1380  
               
               
                   
               
               
                 cgccgtgttt cagcccaata aaaattgaga acgtgtggga cgaatctgat gatggatcga   1440  
               
               
                   
               
               
                 ttagaatcca ggtctcggca caattcggct acaatcaggc aggcactgcg gatgtcacca   1500  
               
               
                   
               
               
                 aattccgtta catgtctttc gaccacgacc atgacatcaa ggaagacagt atggagaaaa   1560  
               
               
                   
               
               
                 tagctatcag cacatctgga ccctgccgtc gtcttggcca caaagggtac ttcctgttag   1620  
               
               
                   
               
               
                 ctcaatgtcc tccaggtgac agtgtaaccg tcagtatcac gagcggagca tctgagaatt   1680  
               
               
                   
               
               
                 catgcaccgt ggagaaaaag atcaggagga agtttgtcgg tagagaggag tacttgttcc   1740  
               
               
                   
               
               
                 cacccgtcca tggaaagctg gtaaagtgcc acgtttacga tcacttgaag gagacgtctg   1800  
               
               
                   
               
               
                 ccgggtacat aaccatgcac aggccaggcc cacacgcgta taagtcctat ctggaggaag   1860  
               
               
                   
               
               
                 cgtcaggcga agtgtacatt aaaccacctt ctggcaagaa cgtcacctac gaatgtaagt   1920  
               
               
                   
               
               
                 gtggcgacta cagcacaggt atcgtgagca cgcgaacgaa gatgaacggc tgcactaaag   1980  
               
               
                   
               
               
                 caaaacagtg cattgcctac aagagcgacc aaacgaaatg ggtcttcaac tcgccggatc   2040  
               
               
                   
               
               
                 ttattaggca cacagaccac tcagtgcaag gtaaattgca cattccattc cgcttgacac   2100  
               
               
                   
               
               
                 cgacagtctg cccggttccg ttagctcaca cgcctacagt cacgaagtgg ttcaaaggca   2160  
               
               
                   
               
               
                 tcaccctcca cctgactgca atgcgaccaa cattgctgac aacgagaaaa ttggggctgc   2220  
               
               
                   
               
               
                 gagcagacgc aacagcagaa tggattacag ggtctacatc caggaatttt tctgtggggc   2280  
               
               
                   
               
               
                 gagaagggct ggagtacgta tggggtaacc atgaaccagt cagagtctgg gcccaggagt   2340  
               
               
                   
               
               
                 cggcaccagg cgacccacat ggatggccgc atgagatcat catccactat tatcatcggc   2400  
               
               
                   
               
               
                 atccagtcta cactgtcatt gtgctgtgtg gtgtcgctct tgctatcctg gtaggcactg   2460  
               
               
                   
               
               
                 catcatcagc agcttgcatc gccaaagcaa gaagagactg cctgacgcca tacgcgcttg   2520  
               
               
                   
               
               
                 caccgaacgc aacggtaccc acagcattag cggttttgtg ctgcattcgg ccaaccaacg   2580  
               
               
                   
               
               
                 ctgaaacatt tggagaaact ttgaaccatc tgtggtttaa caaccaaccg tttctctggg   2640  
               
               
                   
               
               
                 cacagttgtg cattcctctg gcagcgcttg ttattctgtt ccgctgcttt tcatgctgca   2700  
               
               
                   
               
               
                 tgcctttttt attggttgca ggcgtctgcc tggggaaggt agacgccttc gaacatgcga   2760  
               
               
                   
               
               
                 ccactgtgcc aaatgttccg gggatcccgt ataaggcgtt ggtcgaacgc gcaggttacg   2820  
               
               
                   
               
               
                 cgccacttaa cctggagatc acggtcgtct catcggaatt aacaccttca actaacaagg   2880  
               
               
                   
               
               
                 agtacgtgac ctgcaaattc cacacagtca ttccttcacc acaagttaaa tgctgcgggt   2940  
               
               
                   
               
               
                 ccctcgagtg caaggcatcc tcaaaggcgg attacacatg ccgcgttttt ggcggtgtgt   3000  
               
               
                   
               
               
                 accctttcat gtggggaggc gcacaatgct tctgtgacag tgagaacaca caactgagtg   3060  
               
               
                   
               
               
                 aggcgtacgt cgagttcgct ccagactgca ctatagatca cgcagtcgca ctaaaagttc   3120  
               
               
                   
               
               
                 acacagctgc tctgaaagtc ggcctgcgta tagtatacgg caacaccacc gcgcacctgg   3180  
               
               
                   
               
               
                 atacgtttgt caatggcgtc acgccaggtt cctcacggga cctgaaggtc atagcagggc   3240  
               
               
                   
               
               
                 cgatatcagc cgctttttca ccctttgacc ataaggtcgt catcagaaag gggcttgttt   3300  
               
               
                   
               
               
                 acaactacga cttccctgag tatggagcta tgaaaccagg agcgttcggc gatattcaag   3360  
               
               
                   
               
               
                 catcctcgct tgatgctaca gacatagtag cccgcactga catacggctg ctgaagcctt   3420  
               
               
                   
               
               
                 ctgtcaagaa catccacgtc ccctacaccc aagcagtatc agggtatgaa atgtggaaga   3480  
               
               
                   
               
               
                 acaactcagg acgacccctg caagaaacag caccatttgg atgtaaaatt gaagtggagc   3540  
               
               
                   
               
               
                 ctctgcgagc gtctaactgt gcttacgggc acatccctat ctcgattgac atccctgatg   3600  
               
               
                   
               
               
                 cagcttttgt gagatcatca gaatcaccaa caattttaga agttagctgc acagtagcag   3660  
               
               
                   
               
               
                 actgcattta ttctgcagac tttggtggtt ctctaacatt acagtacaaa gctgacaggg   3720  
               
               
                   
               
               
                 agggacattg tccagttcac tcccactcca cgacagctgt tttgaaggaa gcgaccacac   3780  
               
               
                   
               
               
                 atgtgactgc cgtaggcagc ataacactac attttagcac atcgagccca caagcaaatt   3840  
               
               
                   
               
               
                 ttatagtttc gctatgcggc aagaagtcca cctgcaatgc tgaatgtaaa ccaccggccg   3900  
               
               
                   
               
               
                 accacataat tggagaacca cataaagtcg accaagaatt ccaggcggca gtttccaaaa   3960  
               
               
                   
               
               
                 catcttggaa ctggctgctt gcactgtttg ggggagcatc atccctcatt gttgtaggac   4020  
               
               
                   
               
               
                 ttatagtgtt ggtctgcagc tctatgctta taaacacacg tagatgactg agcgcggaca   4080  
               
               
                   
               
               
                 ctgacatagc ggtaaaactc gatgtacttc cgaggaagcg tggtgcataa tgccacgcgc   4140  
               
               
                   
               
               
                 cgcttgacac taaaactcga tgtatttccg aggaagcaca gtgcataatg ctgtgcagtg   4200  
               
               
                   
               
               
                 tcacattaat cgtatatcac actacatatt aacaacacta tatcactttt atgagactca   4260  
               
               
                   
               
               
                 ctatgggtct ctaatataca ctacacatat tttacttaaa aacactatac acactttata   4320  
               
               
                   
               
               
                 aattctttta taatttttct tttgctttag agcacactgg cggccgttac tagtggatcc   4380  
               
               
                   
               
               
                 gagctctaga gggcc                                                    4395