Abstract:
The present invention relates to compositions and methods for enhancing the effect of vaccines in animals, such as domestic, sport, or pet species, and humans. More particularly, the use of Ribavirin as an adjuvant to a vaccine protocol and compositions having Ribavirin and an antigen are described.

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS  
       [0001]    This application is a continuation of U.S. patent application Ser. No. 10/104,966 filed on Mar. 22, 2002, which is a continuation of U.S. patent application Ser. No. 09/705,547 filed on Nov. 3, 2000, both of which claim the benefit of priority of U.S. provisional patent application No. 60/229,175, filed Aug. 29, 2000. The aforementioned applications are hereby expressly incorporated by reference in their entireties. 
     
    
     
       FIELD OF THE INVENTION  
         [0002]    The present invention relates to compositions and methods for enhancing the effect of vaccines in animals, such as domestic, sport, or pet species, and humans. More particularly, preferred embodiments concern the use of Ribavirin as an adjuvant and compositions having Ribavirin and an antigen.  
         BACKGROUND OF THE INVENTION  
         [0003]    The use of vaccines to prevent disease in humans, farm livestock, sports animals, and household pets is a common practice. Frequently, however, the antigen used in a vaccine is not sufficiently immunogenic to raise the antibody titre to levels that are sufficient to provide protection against subsequent challenge or to maintain the potential for mounting these levels over extended time periods. Further, many vaccines are altogether deficient in inducing cell-mediated immunity, which is a primary immune defense against bacterial and viral infection. A considerable amount of research is currently focussed on the development of more potent vaccines and ways to enhance the immunogenicity of antigen-containing preparations. (See e.g., U.S. Pat. Nos. 6,056,961; 6,060,068; 6,063,380; and Li et al.,  Science  288:2219-2222 (2000)).  
           [0004]    Notorious among such “weak” vaccines are hepatitis B vaccines. For example, recombinant vaccines against hepatitis B virus such as Genhevacb (Pasteur Merieux Serums et Vaccines, 58, Avenue Leclerc 69007 Lyon, France), Engerixb (Smith, Kline and Symbol French), and Recombivaxhb (Merck, Sharp, and Dhome) are effective only after at least three injections at 0, 30, and 60 or 180 days, followed by an obligatory booster after one year. (Chedid et al., U.S. Pat. No. 6,063,380). Additionally, many subjects receiving these vaccines respond poorly, if at all. Because many regions of the world are endemic for HBV infection, the poorly immunogenic character of existing HBV vaccines has become an extremely serious problem.  
           [0005]    To obtain a stronger, humoral and/or cellular response, it is common to administer a vaccine in a material that enhances the immune response of the patient to the antigen present in the vaccine. The most commonly used adjuvants for vaccine protocols are oil preparations and alum. (Chedid et al., U.S. Pat. No. 6,063,380). A greater repertoire of safe and effective adjuvants is needed.  
           [0006]    Nucleoside analogs have been widely used in anti-viral therapies due to their capacity to reduce viral replication. (Hosoya et al.,  J. Inf. Dis.,  168:641-646 (1993)). Ribavirin (1-β-D-ribofuranosyl-1,2,4-triazole-3-carboxamide) is a synthetic guanosine analog that has been used to inhibit RNA and DNA virus replication. (Huffman et al.,  Antimicrob. Agents. Chemother.,  3:235 (1973); Sidwell et al.,  Science,  177:705 (1972)). Ribavirin has been shown to be a competitive inhibitor of inositol mono-phosphate (IMP) dehydrogenase (IMPDH), which converts IMP to IMX (which is then converted to GMP). De Clercq,  Anti viral Agents: characteristic activity spectrum depending on the molecular target with which they interact,  Academic press, Inc., New York N.Y., pp. 1-55 (1993). Intracellular pools of GTP become depleted as a result of long term Ribavirin treatment.  
           [0007]    In addition to antiviral activity, investigators have observed that a few guanosine analogs have an effect on the immune system. (U.S. Pat. Nos. 6,063,772 and 4,950,647). Ribavirin has been shown to inhibit functional humoral immune responses (Peavy et al.,  J. Immunol.,  126:861-864 (1981); Powers et al.,  Antimicrob. Agents. Chemother.,  22:108-114 (1982)) and IgE-mediated modulation of mast cell secretion. (Marquardt et al.,  J. Pharmacol. Exp. Therapeutics,  240:145-149 (1987)). Some investigators report that a daily oral therapy of Ribavirin has an immune modulating effect on humans and mice. (Hultgren et al.,  J. Gen. Virol.,  79:2381-2391 (1998) and Cramp et al.,  Gastron. Enterol.,  118:346-355 (2000)). Nevertheless, the current understanding of the effects of Ribavirin on the immune system is in its infancy.  
         SUMMARY OF THE INVENTION  
         [0008]    It has been discovered that Ribavirin can be used as an adjuvant to enhance an immune response to an antigen. Embodiments described herein include “strong” vaccine preparations that comprise an antigen and Ribavirin. Generally, these preparations have an amount of Ribavirin that is sufficient to enhance an immune response to the antigen. Other aspects of the invention include methods of enhancing the immune response of an animal, including a human, to an antigen. By one approach, for example, an animal in need of a potent immune response to an antigen is identified and then is provided an amount of Ribavirin together with the antigen that is effective to enhance an immune response in the animal. In some methods, the Ribavirin and the antigen are provided in combination and in others, the Ribavirin and the antigen are provided separately. Thus, several embodiments concern the manufacture and use of vaccine preparations having Ribavirin and an antigen.  
           [0009]    Preferred vaccine compositions comprise Ribavirin and a hepatitis viral antigen. The antigen can be a peptide or nucleic acid-based (e.g., a RNA encoding a peptide antigen or a construct that expresses a peptide antigen when introduced to a subject). HBV antigens that are suitable include, for example, hepatitis B surface antigen (HBsAg), hepatitis core antigen (HBcAg), hepatitis e antigen (HBeAg), and nucleic acids encoding these molecules. Compositions having Ribavirin and an antigen from the hepatitis A virus (HAV) or Ribavirin and a nucleic acid encoding an antigen from HAV are also embodiments. Still further, compositions having Ribavirin and an antigen from the hepatitis C virus (HCV) or Ribavirin and a nucleic acid encoding an antigen from HCV are embodiments.  
           [0010]    Furthermore, compositions having a mixture of the antigens above are embodiments of the present invention. For example, some compositions comprise a HBV antigen, a HAV antigen, and Ribavirin or a HBV antigen, a HCV antigen, and Ribavirin or a HAV antigen, a HCV antigen, and Ribavirin or a HBV antigen, a HAV antigen, a HCV antigen, and Ribavirin. Other embodiments comprise Ribavirin and a nucleic acid encoding a mixture of the antigens described above. Some embodiments also include other adjuvants, binders, emulsifiers, carriers, and fillers, as known in the art, including, but not limited to, alum, oil, and other compounds that enhance an immune response.  
           [0011]    Preferred methods involve providing an animal in need with a sufficient amount of Ribavirin and a hepatitis viral antigen (e.g., HBV antigen, HAV antigen, HCV antigen a nucleic acid encoding one of these antigens or any combination thereof). Accordingly, one embodiment includes identifying an animal in need of an enhanced immune response to a hepatitis viral antigen (e.g., an animal at risk or already infected with a hepatitis infection) and providing to said animal an amount of Ribavirin that is effective to enhance an immune response to the hepatitis viral antigen. 
       
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0012]    [0012]FIG. 1 is a graph showing the humoral response to 10 and 100 μg recombinant Hepatitis C virus (HCV) non structural 3 protein (NS3), as determined by mean end point titres, when a single dose of 1 mg of Ribavirin was co-administered.  
         [0013]    [0013]FIG. 2 is a graph showing the humoral response to 20 μg recombinant Hepatitis C virus (HCV) non structural 3 protein (NS3), as determined by mean end point titres, when a single dose of 0.1, 1.0, or 10 mg of Ribavirin was co-administered.  
         [0014]    [0014]FIG. 3 is a graph showing the effects of a single dose of 1 mg Ribavirin on NS3-specific lymph node proliferative responses, as determined by in vitro recall responses. 
     
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0015]    It has been discovered that compositions comprising Ribavirin and an antigen can boost an animal&#39;s immune response to the antigen. That is, Ribavirin can be used as an “adjuvant,” which for the purposes of this disclosure, refers to a compound that has the ability to enhance the immune response to a particular antigen. Such adjuvant activity is manifested by a significant increase in immune-mediated protection against the antigen, and was demonstrated by an increase in the titer of antibody raised to the antigen and an increase in proliferative T cell responses.  
         [0016]    Several vaccine preparations that comprise Ribavirin and an antigen are described herein. Vaccine formulations containing Ribavirin can vary according to the amount of Ribavirin, the form of Ribavirin, and the type of antigen. The antigen can be a peptide or a nucleic acid (e.g., a RNA encoding a peptide antigen or a construct that expresses a peptide antigen when introduced into a subject). Preferred vaccine formulations comprise Ribavirin and a hepatitis viral antigen (e.g., HBV antigen, HAV antigen, HCV antigen, a nucleic acid encoding these molecules, or any combination thereof).  
         [0017]    Methods of enhancing the immune response of an animal, including humans, to an antigen are also described herein. One method, for example, involves identifying an animal in need of an enhanced immune response to an antigen and providing the animal the antigen and an amount of Ribavirin that is effective to enhance an immune response to the antigen. Preferred methods involve providing the animal in need with Ribavirin and a hepatitis antigen (e.g., HBV antigen, HAV antigen, HCV antigen, a nucleic acid encoding these molecules, or any combination thereof). The section below describes the manufacture of vaccines having Ribavirin and an antigen in greater detail.  
         [0018]    Vaccines Containing Ribavirin  
         [0019]    The vaccines comprise Ribavirin and an antigen and may contain other ingredients including, but not limited to, adjuvants, binding agents, excipients such as stabilizers (to promote long term storage), emulsifiers, thickening agents, salts, preservatives, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. These vaccine preparations are suitable for treatment of animals either as a preventive measure to avoid a disease or condition or as a therapeutic to treat animals already afflicted with a disease or condition.  
         [0020]    The vaccine compositions can be manufactured in accordance with conventional methods of galenic pharmacy to produce medicinal agents for administration to animals, e.g., mammals including humans. Ribavirin can be obtained from commercial suppliers (e.g., Sigma and ICN). Ribavirin and/or the antigen can be formulated into the vaccine with and without modification. For example, the Ribavirin and/or antigen can be modified or derivatized to make a more stable molecule and/or a more potent adjuvant. By one approach, the stability of Ribavirin and/or an antigen can be enhanced by coupling the molecules to a support such as a hydrophilic polymer (e.g., polyethylene glycol).  
         [0021]    Many more Ribavirin derivatives can be generated using conventional techniques in rational drug design and combinatorial chemistry. For example, Molecular Simulations Inc. (MSI), as well as many other suppliers, provide software that allows one of skill to build a combinatorial library of organic molecules. The C2.Analog Builder program, for example, can be integrated with MSI&#39;s suite of Cerius2 molecular diversity software to develop a library of Ribavirin derivatives that can be used with the embodiments described herein. (See e.g., http://msi.com/life/products/cerius2/index.html, herein expressly incorporated by reference in its entirety).  
         [0022]    By one approach, the chemical structure of Ribavirin is recorded on a computer readable media and is accessed by one or more modeling software application programs. The C2.Analog Builder program in conjunction with C2Diversity program allows the user to generate a very large virtual library based on the diversity of R-groups for each substituent position, for example. Compounds having the same structure as the modeled Ribavirin derivatives created in the virtual library are then made using conventional chemistry or can be obtained from a commercial source.  
         [0023]    The newly manufactured Ribavirin derivatives are then screened in characterization assays, which determine the extent of adjuvant activity of the molecule and/or the extent of its ability to modulate of an immune response. Some characterization assays may involve virtual drug screening software, such as C2.Ludi. C2.Ludi is a software program that allows a user to explore databases of molecules (e.g., Ribavirin derivatives) for their ability to interact with the active site of a protein of interest (e.g., RAC2 or another GTP binding protein). Based upon predicted interactions discovered with the virtual drug screening software, the Ribavirin derivatives can be prioritized for further characterization in conventional assays that determine adjuvant activity and/or the extent of a molecule to modulate an immune response.  
         [0024]    Example 1 describes a characterization assay that was used to evaluate the adjuvant activity of Ribavirin.  
       EXAMPLE 1  
       [0025]    This characterization assay can be used with any Ribavirin derivative or combinations of Ribavirin derivatives to determine the extent of adjuvant activity of the particular vaccine formulation. Accordingly, groups of three to five Balb/c mice (BK Universal, Uppsala, Sweden) were immunized i.p or s.c. (e.g., at the base of the tail) with 10 μg or 100 μg of recombinant hepatitis C virus non-structural 3 (NS3) protein. The rNS3 was dissolved in phosphate buffered saline (PBS) alone or PBS containing 1 mg Ribavirin (obtained from ICN, Costa Mesa, Calif.). Mice were injected with a total volume of 100 μl per injection.  
         [0026]    At two and four weeks following ip. immunization, all mice were bled by retro-orbital sampling. Serum samples were collected and analyzed for the presence of antibodies to rNS3. To determine the antibody titer, an enzyme immunoassay (EIA) was performed. (See e.g., Hultgren et al.,  J Gen Virol.  79:2381-91 (1998) and Hultgren et al.,  Clin. Diagn. Lab. Immunol.  4:630-632 (1997), both of which are herein expressly incorporated by reference in their entireties). The antibody levels were recorded as the highest serum dilution giving an optical density at 405 nm more than twice that of non-immunized mice.  
         [0027]    Mice that received 10 μg or 100 μg rNS3 mixed with 1 mg Ribavirin in PBS displayed consistently higher levels of NS3 antibodies. The antibody titer that was detected by EIA at two weeks post-immunization is shown in FIG. 1. The vaccine formulations having 1 mg of Ribavirin and either 10 μg or 100 μg of rNS3 induced a significantly greater antibody titer than the vaccine formulations composed of only rNS3. This data provides evidence that Ribavirin has an adjuvant effect on the humoral immune response of an animal and thus, enhances the immune response to the antigen.  
         [0028]    The example below describes experiments that were performed to determine the amount of Ribavirin that was needed to elicit an adjuvant effect.  
       EXAMPLE 2  
       [0029]    To determine the dose of Ribavirin that is required to provide an adjuvant effect, the following experiments were performed. Groups of mice (three per group) were immunized with a 20 μg rNS3 alone or a mixture of 20 μg rNS3 and 0.1 mg, 1 mg, or 10 mg Ribavirin. The levels of antibody to the antigen were then determined by EIA. The mean endpoint titers at weeks 1 and 3 were plotted and are shown in FIG. 2. It was discovered that the adjuvant effect provided by Ribavirin had different kinetics depending on the dose of Ribavirin provided. For example, low doses (&lt;1 mg) of Ribavirin were found to enhance antibody levels at week one but not at week three, whereas, higher doses (1-10 mg) were found to enhance antibody levels at week three. These data further verify that Ribavirin can be administered as an adjuvant and establish that that the dose of Ribavirin can modulate the kinetics of the adjuvant effect.  
         [0030]    The example below describes another characterization assay that was performed to evaluate the ability of Ribavirin to modulate a cellular immune response.  
       EXAMPLE 3  
       [0031]    This characterization assay can be used with any Ribavirin derivative or combinations of Ribavirin derivatives to determine the extent that a particular vaccine formulation modulates a cellular immune response. To determine CD4 +  T cell responses to Ribavirin-containing vaccine, groups of mice were immunized s.c. with either 100 μg rNS3 in PBS or 100μg rNS3 and 1 mg Ribavirin in PBS. The mice were sacrificed ten days post-immunization and their lymph nodes were harvested and drained. In vitro recall assays were then performed. (See e.g., Hultgren et al.,  J Gen Virol.  79:2381-91 (1998) and Hultgren et al.,  Clin. Diagn. Lab. Immunol.  4:630-632 (1997), both of which are herein expressly incorporated by reference in their entireties). The amount of CD4 +  T cell proliferation was determined at 96 h of culture by the incorporation of [ 3 H] thymidine.  
         [0032]    As shown in FIG. 2, mice that were immunized with 100 μg rNS3 mixed with 1 mg Ribavirin had a much greater T cell proliferative response than mice that were immunized with 100 μg rNS3 in PBS. This data provides evidence that Ribavirin can enhance a cellular immune response (e.g., by promoting the effective priming of T cells).  
         [0033]    The example below describes the use of Ribavirin in conjunction with a commercial vaccine preparation.  
       EXAMPLE 4  
       [0034]    The adjuvant effect of Ribavirin was also tested when mixed with two doses of a commercially available vaccine containing HBsAg and alum. (Engerix, SKB). Approximately 0.2 μg or 2 μg of Engerix vaccine was mixed with either PBS or 1 mg Ribavirin in PBS and the mixtures were injected intra peritoneally into groups of mice (three per group). A booster containing the same mixture was given on week four and all mice were bled on week six. The serum samples were diluted from 1:60 to 1:37500 and the dilutions were tested by EIA, as described above, except that purified human HBsAg (kindly provided by Professor DL Peterson, Commonwealth University, Va.) was used as the solid phase antigen. As shown in TABLE 1, vaccine formulations having Ribavirin enhanced the response to 2 μg of an existing vaccine despite the fact that the vaccine already contained alum. That is, by adding Ribavirin to a suboptimal vaccine dose (i.e., one that does not induce detectable antibodies alone) antibodies became detectable, providing evidence that the addition of Ribavirin allows for the use of lower antigen amounts in a vaccine formulation without compromising the immune response.  
                                                                                                         TABLE 1                           End point antibody titer to HBsAg in EIA                0.02 μg Engerix   0.2 μg Engerix                No Ribavirin   1 mg Ribavirin   No Ribavirin   1 mg Ribavirin            Week   #1   #2   #3   #1   #2   #3   #1   #2   #3   #1   #2   #3               6   &lt;60   &lt;60   &lt;60   &lt;60   &lt;60   &lt;60   &lt;60   &lt;60   &lt;60   300   60   &lt;60                  
 
         [0035]    Any antigen that can be used to generate an immune response in an animal can be combined with Ribavirin so as to prepare the vaccines described herein. That is, antigens that can be incorporated into such a vaccine comprise bacterial antigens, fungal antigens, plant antigens, mold antigens, viral antigens, cancer cell antigens, toxin antigens, chemical antigens, and self-antigens. Although many of these antigens are molecules that induce a significant immune response without an adjuvant, Ribavirin can be administered in conjunction with or combined with “strong” or “weak” antigens to enhance the immune response. In addition, the use of Ribavirin as an adjuvant may allow for the use of lower amounts of vaccine antigens while retaining immunogenicity.  
         [0036]    Preferred embodiments comprise Ribavirin and a viral antigen. Preferred viral antigens are hepatitis viral antigens. Vaccines can comprise, for example, Ribavirin and an HBV antigen, HAV antigen, HCV antigen or any combination of these antigens. Preferred viral antigens include hepatitis B surface antigen (HBsAg), hepatitis core antigen (HBcAg), and hepatitis E antigen (HBeAg).  
         [0037]    For example, HCV vaccine embodiments comprise Ribavirin and a HCV peptide of at least 3 consecutive amino acids of SEQ. ID. No.: 1. That is, a vaccine embodiment can have Ribavirin and a HCV peptide with a length of at least 3-10 consecutive amino acids, 10-50 consecutive amino acids, 50-100 consecutive amino acids, 100-200 consecutive amino acids, 200-400 consecutive amino acids, 400-800 consecutive amino acids, 800-1200 consecutive amino acids, 1200-1600 consecutive amino acids, 1600-2000 consecutive amino acids, 2000-2500 consecutive amino acids, and 2500-3011 consecutive amino acids of SEQ ID. No. 1. Preferred HCV vaccines comprise Ribavirin and a peptide of at least 3 consecutive amino acids of HCV core protein (SEQ. ID. No. 2), HCV E1 protein (SEQ. ID. No. 3), HCV E2 protein (SEQ. ID. No. 4), HCV NS2 (SEQ. ID. No. 5), HCV NS3 (SEQ. ID. No. 6), HCV NS4A (SEQ. ID. No. 7), HCV NS4B (SEQ. ID. No. 8), or HCV NS5A/B (SEQ. ID. No. 9). That is, preferred HCV vaccines can comprise Ribavirin and a peptide with a length of at least 3-10 consecutive amino acids, 10-50 consecutive amino acids, 50-100 consecutive amino acids, 100-200 consecutive amino acids, 200-400 consecutive amino acids, 400-800 consecutive amino acids, and 800-1040 consecutive amino acids of any one of (SEQ. ID. Nos. 2-9).  
         [0038]    Similarly, preferred HBV vaccine embodiments comprise Ribavirin and a HBV peptide of at least 3 consecutive amino acids of HBsAg (SEQ. ID. No.: 10) or HBcAg and HBeAg (SEQ. ID. No. 11). That is, a vaccine embodiment can have Ribavirin and a HBV peptide with a length of at least 3-10 consecutive amino acids, 10-50 consecutive amino acids, 50-100 consecutive amino acids, 100-150 consecutive amino acids, 150-200 consecutive amino acids, and 200-226 consecutive amino acids of either SEQ. ID. No. 10 or SEQ. ID. No. 11. Further, preferred HAV embodiments comprise Ribavirin and a HAV peptide with a length of at least 3-10 consecutive amino acids, 10-50 consecutive amino acids, 50-100 consecutive amino acids, 100-200 consecutive amino acids, 200-400 consecutive amino acids, 400-800 consecutive amino acids, 800-1200 consecutive amino acids, 1200-1600 consecutive amino acids, 1600-2000 consecutive amino acids, and 2000-2227 consecutive amino acids of SEQ ID. No. 12.  
         [0039]    In addition to peptide antigens, nucleic acid-based antigens can be used in the vaccine compositions described herein. Various nucleic acid-based vaccines are known and it is contemplated that these compositions and approaches to immunotherapy can be augmented by introducing Ribavirin (See e.g., U.S. Pat. No. 5,589,466, herein expressly incorporated by reference in its entirety).  
         [0040]    By one approach, for example, a gene encoding a polypeptide antigen of interest is cloned into an expression vector capable of expressing the polypeptide when introduced into a subject. The expression construct is introduced into the subject in a mixture of Ribavirin or in conjunction with Ribavirin (e.g., Ribavirin is administered shortly after the expression construct at the same site). Alternatively, RNA encoding a polypeptide antigen of interest is provided to the subject in a mixture with Ribavirin or in conjunction with Ribavirin. Where the polynucleotide is to be DNA, promoters suitable for use in various vertebrate systems are well known. For example, for use in murine systems, suitable strong promoters include RSV LTR, MPSV LTR, SV40 IEP, and metallothionein promoter. In humans, on the other hand, promoters such as CMV IEP can be used. All forms of DNA, whether replicating or non-replicating, which do not become integrated into the genome, and which are expressible, can be used.  
         [0041]    Preferred nucleic acid-based antigens include a nucleotide sequence of at least 9 consecutive nucleotides of HCV (SEQ. ID. No. 13), HBV (SEQ. ID. No.:14), or HAV (SEQ. ID. No. 15). That is, a nucleic acid based antigen can comprise at least 9-25 consecutive nucleotides, 25-50 consecutive nucleotides, 50-100 consecutive nucleotides, 100-200 consecutive nucleotides, 200-500 consecutive nucleotides, 500-1000 consecutive nucleotides, 1000-2000 consecutive nucleotides, 2000-4000 consecutive nucleotides, 4000-8000 consecutive nucleotides, and 8000-9416 consecutive nucleotides of any one of SEQ. ID. Nos.: 13-15 or an RNA that corresponds to these sequences.  
         [0042]    The example below describes one approach for using a nucleic acid-based antigen in conjunction with Ribavirin.  
       EXAMPLE 5  
       [0043]    The following describes an approach to immunize an animal with a vaccine comprising a nucleic acid-based antigen and Ribavirin. Five to six week old female and male Balb/C mice are anesthetized by intraperitoneal injection with 0.3 ml of 2.5% Avertin. A 1.5 cm incision is made on the anterior thigh, and the quadriceps muscle is directly visualized. One group of mice are injected with approximately 20 μg of an expression construct having the gp-120 gene, driven by a cytomegalovirus (CMV) promotor and second group of mice are injected with approximately 5 μg of capped in vitro transcribed RNA (e.g., SP6, T7, or T3 (Ambion)) encoding gp-120. These two groups are controls. A third group of mice is injected with approximately 20 μg of the expression vector having the gp-120 gene and the CMV promoter mixed with 1 mg of Ribavirin and a fourth group of mice is injected with approximately 5 μg of capped in vitro transcribed RNA mixed with 1 mg Rbavirin. The vaccines are injected in 0.1 ml of solution (PBS) in a 1 cc syringe through a 27 gauge needle over one minute, approximately 0.5 cm from the distal insertion site of the muscle into the knee and about 0.2 cm deep. A suture is placed over the injection site for future localization, and the skin is then closed with stainless steel clips.  
         [0044]    Blood samples are obtained prior to the injection (Day 0) and up to more than 40 days post injection. The serum from each sample is serially diluted and assayed in a standard ELISA technique assay for the detection of antibody, using recombinant gp-120 protein made in yeast as the antigen. Both IgG and IgM antibodies specific for gp-120 will be detected in all samples, however, groups three and four, which contained the Ribavirin, will exhibit a greater immune response to the gp-120 as measured by the amount and/or titer of antibody detected in the sera.  
         [0045]    Many other ingredients can be present in the vaccine. For example, the Ribavirin and antigen can be employed in admixture with conventional excipients (e.g., pharmaceutically acceptable organic or inorganic carrier substances suitable for parenteral, enteral (e.g., oral) or topical application that do not deleteriously react with the Ribavirin and/or antigen). Suitable pharmaceutically acceptable carriers include, but are not limited to, water, salt solutions, alcohols, gum arabic, vegetable oils, benzyl alcohols, polyetylene glycols, gelatine, carbohydrates such as lactose, amylose or starch, magnesium stearate, talc, silicic acid, viscous paraffin, perfume oil, fatty acid monoglycerides and diglycerides, pentaerythritol fatty acid esters, hydroxy methylcellulose, polyvinyl pyrrolidone, etc. Many more suitable carriers are described in Remmington&#39;s Pharmaceutical Sciences, 15th Edition, Easton:Mack Publishing Company, pages 1405-1412 and 1461-1487(1975) and The National Formulary XIV, 14th Edition, Washington, American Pharmaceutical Association (1975), herein expressly incorporated by reference in their entireties. Vaccines can be sterilized and if desired mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, flavoring and/or aromatic substances and the like that do not deleteriously react with Ribavirin or the antigen.  
         [0046]    The effective dose and method of administration of a particular vaccine formulation can vary based on the individual patient and the type and stage of the disease, as well as other factors known to those of skill in the art. Therapeutic efficacy and toxicity of the vaccines can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., ED50 (the dose therapeutically effective in 50% of the population). The data obtained from cell culture assays and animal studies can be used to formulate a range of dosage for human use. The dosage of the vaccines lies preferably within a range of circulating concentrations that include the ED50 with no toxicity. The dosage varies within this range depending upon the type of Ribavirin derivative and antigen, the dosage form employed, the sensitivity of the patient, and the route of administration.  
         [0047]    Since Ribavirin has been on the market for several years, many dosage forms and routes of administration are known. All known dosage forms and routes of administration can be provided within the context of the embodiments described herein. Preferably, an amount of Ribavirin that is effective to enhance an immune response to an antigen in an animal can be considered to be an amount that is sufficient to achieve a blood serum level of antigen approximately 0.25-12.5 μg/ml in the animal, preferably, about 2.5 μg/ml. In some embodiments, the amount of Ribavirin is determined according to the body weight of the animal to be given the vaccine. Accordingly, the amount of Ribavirin in a vaccine formulation can be from about 0.1-6.0 mg/kg body weight. That is, some embodiments have an amount of Ribavirin that corresponds to approximately 0.1-1.0 mg/kg, 1.1-2.0 mg/kg, 2.1-3.0 mg/kg, 3.1-4.0 mg/kg, 4.1-5.0 mg/kg, 5.1, and 6.0 mg/kg body weight of an animal. More conventionally, the vaccines contain approximately 0.25 mg-2000 mg of Ribavirin. That is, some embodiments have approximately 250 μg, 500 μg, 1 mg, 25 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 1 g, 1.1 g, 1.2 g, 1.3 g, 1.4 g, 1.5 g, 1.6 g, 1.7 g, 1.8 g, 1.9 g, and 2 g of Ribavirin.  
         [0048]    Vaccines comprising various antigens and amounts of these antigens have been provided to animals for many years. Thus, conventional vaccine preparations can be modified by adding an amount of Ribavirin that is sufficient to enhance an immune response to the antigen. That is, existing conventional vaccine formulations can be modified by simply adding Ribavirin to the preparation or by administering the conventional vaccine in conjunction with Ribavirin (e.g., shortly before or after providing the antigen). As one of skill in the art will appreciate, the amount of antigens in a vaccine can vary depending on the type of antigen and its immunogenicity. The amount of antigens in the vaccines can vary accordingly. Nevertheless, as a general guide, the vaccines can have approximately 0.25 mg-2000 mg of an antigen (e.g., a hepatitis viral antigen).  
         [0049]    In some approaches described herein, the exact amount of Ribavirin and/or antigen is chosen by the individual physician in view of the patient to be treated. Further, the amounts of Ribavirin can be added in combination to or separately from the same or equivalent amount of antigen and these amounts can be adjusted during a particular vaccination protocol so as to provide sufficient levels in light of patient-specific or antigen-specific considerations. In this vein, patient-specific and antigen-specific factors that can be taken into account include, but are not limited to, the severity of the disease state of the patient, age, and weight of the patient, diet, time and frequency of administration, drug combination(s), reaction sensitivities, and tolerance/response to therapy.  
         [0050]    Routes of administration of the vaccines described herein include, but are not limited to, transdermal, parenteral, gastrointestinal, transbronchial, and transalveolar. Transdermal administration can be accomplished by application of a cream, rinse, gel, etc. capable of allowing Ribavirin and antigen to penetrate the skin. Parenteral routes of administration include, but are not limited to, electrical or direct injection such as direct injection into a central venous line, intravenous, intramuscular, intraperitoneal, intradermal, or subcutaneous injection. Gastrointestinal routes of administration include, but are not limited to, ingestion and rectal. Transbronchial and transalveolar routes of administration include, but are not limited to, inhalation, either via the mouth or intranasally.  
         [0051]    Compositions having Ribavirin and an antigen that are suitable for transdermal administration include, but are not limited to, pharmaceutically acceptable suspensions, oils, creams, and ointments applied directly to the skin or incorporated into a protective carrier such as a transdermal device (“transdermal patch”). Examples of suitable creams, ointments, etc. can be found, for instance, in the Physician&#39;s Desk Reference. Examples of suitable transdermal devices are described, for instance, in U.S. Pat. No. 4,818,540 issued Apr. 4, 1989 to Chinen, et al., herein expressly incorporated by reference in its entirety.  
         [0052]    Compositions having Ribavirin and an antigen that are suitable for parenteral administration include, but are not limited to, pharmaceutically acceptable sterile isotonic solutions. Such solutions include, but are not limited to, saline, phosphate buffered saline and oil preparations for injection into a central venous line, intravenous, intramuscular, intraperitoneal, intradermal, or subcutaneous injection.  
         [0053]    Compositions having Ribavirin and an antigen that are suitable for transbronchial and transalveolar administration include, but not limited to, various types of aerosols for inhalation. Devices suitable for transbronchial and transalveolar administration of these are also embodiments. Such devices include, but are not limited to, atomizers and vaporizers. Many forms of currently available atomizers and vaporizers can be readily adapted to deliver vaccines having Ribavirin and an antigen.  
         [0054]    Compositions having Ribavirin and an antigen that are suitable for gastrointestinal administration include, but not limited to, pharmaceutically acceptable powders, pills or liquids for ingestion and suppositories for rectal administration.  
         [0055]    Once the vaccine comprising Ribavirin and an antigen has been obtained, it can be administered to a subject in need to treat or prevent diseases. The next section describes methods that employ the vaccines described above.  
         [0056]    Methods of Use of Vaccines that Contain Ribavirin  
         [0057]    The vaccines containing Ribavirin and an antigen can be used to treat and prevent a vast spectrum of diseases and can enhance the immune response of an animal to an antigen. As one of skill in the art will appreciate conventional vaccines have been administered to subjects in need of treatment or prevention of bacterial diseases, viral diseases, fungal diseases, and cancer. Because the vaccines described herein include conventional vaccines, which have been modified by the addition of Ribavirin, the methods described herein include the treatment and prevention of a disease using a vaccine that comprises an antigen and Ribavirin.  
         [0058]    Preferred embodiments concern methods of treating or preventing hepatitis infection. In these embodiments, an animal in need is provided a hepatitis antigen (e.g., a peptide antigen or nucleic acid-based antigen) and an amount of Ribavirin sufficient to exhibit an adjuvant activity in said animal. Accordingly, an animal can be identified as one in need by using currently available diagnostic testing or clinical evaluation. The range of hepatitis viral antigens that can be used with these embodiments is diverse. Preferred hepatitis viral antigens include an HBV antigen, an HAV antigen, an HCV antigen, nucleic acids encoding these antigens, or any combination thereof. Highly preferred embodiments include an HBV antigen selected from the group consisting of hepatitis B surface antigen (HBsAg), hepatitis core antigen (HBcAg), and hepatitis E antigen (HBeAg), in particular, the peptide and nucleic acid-based antigens describes supra. The Ribavirin and antigen can be provided separately or in combination, and other adjuvants (e.g., oil, alum, or other agents that enhance an immune response) can also be provided to the animal in need. Thus, preferred embodiments include methods of treating or preventing hepatitis in an animal (e.g., HBV) by identifying an infected animal or an animal at risk of infection and providing said animal a hepatitis antigen (e.g., HBsAg, HBcAg, and HBeAg) and an amount of Ribavirin sufficient to exhibit adjuvant activity.  
         [0059]    Other embodiments include methods of enhancing an immune response to an antigen by providing an animal in need with an amount of Ribavirin that is effective to enhance said immune response. In these embodiments, an animal in need of an enhanced immune response to an antigen is identified by using currently available diagnostic testing or clinical evaluation. Oftentimes these individuals will be suffering from a disease (e.g., bacterial, fungal, mold, viral, or cancer) or are at risk from contracting the disease. However, an animal in need of an enhanced immune response can be an animal that has been poisoned (e.g., bit by a poisonous insect or animal) or that has been exposed to a toxin or other toxic compound. Once identified, these animals are provided an appropriate antigen and an amount of Ribavirin effective to enhance an immune response in the animal.  
         [0060]    As above, the hepatitis viral antigens that can be used with these embodiments include, but are not limited to, an HBV antigen, an HAV antigen, an HCV antigen, a nucleic acid encoding these molecules, or any combination thereof. Highly preferred embodiments include an HBV antigen selected from the group consisting of hepatitis B surface antigen (HBsAg), hepatitis core antigen (HBcAg), and hepatitis E antigen (HBeAg), in particular, the peptide and nucleic acid-based antigens described supra. The Ribavirin and antigen can be provided separately or in combination, and other adjuvants (e.g., oil, alum, or other agents that enhance an immune response) can also be provided to the animal in need. Thus, preferred embodiments include methods of enhancing an immune response to a hepatitis antigen (e.g., HBV) by identifying an animal in need and providing the animal a hepatitis antigen (e.g., HBsAg, HBcAg, and HBeAg) and an amount of Ribavirin that is effective to enhance an immune response in the animal.  
         [0061]    Although the invention has been described with reference to embodiments and examples, it should be understood that various modifications can be made without departing from the spirit of the invention. Accordingly, the invention is limited only by the following claims. All references cited herein are hereby expressly incorporated by reference.  
     
       
       
         1 
         
           
             15  
           
           
             1  
             3011  
             PRT  
             Artificial Sequence  
             
               Hepatitis C virus sequence  
             
           
            1 

Met Ser Thr Asn Pro Lys Pro Gln Arg Lys Thr Lys Arg Asn Thr Asn 
 1               5                  10                  15 

Arg Arg Pro Gln Asp Val Lys Phe Pro Gly Gly Gly Gln Ile Val Gly 
            20                  25                  30 

Gly Val Tyr Leu Leu Pro Arg Arg Gly Pro Arg Leu Gly Val Arg Ala 
        35                  40                  45 

Thr Arg Lys Thr Ser Glu Arg Ser Gln Pro Arg Gly Arg Arg Gln Pro 
    50                  55                  60 

Ile Pro Lys Ala Arg Arg Pro Glu Gly Arg Thr Trp Ala Gln Pro Gly 
65                  70                  75                  80 

Tyr Pro Trp Pro Leu Tyr Gly Asn Glu Gly Cys Gly Trp Ala Gly Trp 
                85                  90                  95 

Leu Leu Ser Pro Arg Gly Ser Arg Pro Ser Trp Gly Pro Thr Asp Pro 
            100                 105                 110 

Arg Arg Arg Ser Arg Asn Leu Gly Lys Val Ile Asp Thr Leu Thr Cys 
        115                 120                 125 

Gly Phe Ala Asp Leu Met Gly Tyr Ile Pro Leu Val Gly Ala Pro Leu 
    130                 135                 140 

Gly Gly Ala Ala Arg Ala Leu Ala His Gly Val Arg Val Leu Glu Asp 
145                 150                 155                 160 

Gly Val Asn Tyr Ala Thr Gly Asn Leu Pro Gly Cys Ser Phe Ser Ile 
                165                 170                 175 

Phe Leu Leu Ala Leu Leu Ser Cys Leu Thr Val Pro Ala Ser Ala Tyr 
            180                 185                 190 

Gln Val Arg Asn Ser Ser Gly Leu Tyr His Val Thr Asn Asp Cys Pro 
        195                 200                 205 

Asn Ser Ser Val Val Tyr Glu Ala Ala Asp Ala Ile Leu His Thr Pro 
    210                 215                 220 

Gly Cys Val Pro Cys Val Arg Glu Gly Asn Ala Ser Arg Cys Trp Val 
225                 230                 235                 240 

Ala Val Thr Pro Thr Val Ala Thr Arg Asp Gly Lys Leu Pro Thr Thr 
                245                 250                 255 

Gln Leu Arg Arg His Ile Asp Leu Leu Val Gly Ser Ala Thr Leu Cys 
            260                 265                 270 

Ser Ala Leu Tyr Val Gly Asp Leu Cys Gly Ser Val Phe Leu Val Gly 
        275                 280                 285 

Gln Leu Phe Thr Phe Ser Pro Arg His His Trp Thr Thr Gln Asp Cys 
    290                 295                 300 

Asn Cys Ser Ile Tyr Pro Gly His Ile Thr Gly His Arg Met Ala Trp 
305                 310                 315                 320 

Asn Met Met Met Asn Trp Ser Pro Thr Ala Ala Leu Val Val Ala Gln 
                325                 330                 335 

Leu Leu Arg Ile Pro Gln Ala Ile Met Asp Met Ile Ala Gly Ala His 
            340                 345                 350 

Trp Gly Val Leu Ala Gly Ile Lys Tyr Phe Ser Met Val Gly Asn Trp 
        355                 360                 365 

Ala Lys Val Leu Val Val Leu Leu Leu Phe Ala Gly Val Asp Ala Glu 
    370                 375                 380 

Thr His Val Thr Gly Gly Asn Ala Gly Arg Thr Thr Ala Gly Leu Val 
385                 390                 395                 400 

Gly Leu Leu Thr Pro Gly Ala Lys Gln Asn Ile Gln Leu Ile Asn Thr 
                405                 410                 415 

Asn Gly Ser Trp His Ile Asn Ser Thr Ala Leu Asn Cys Asn Glu Ser 
            420                 425                 430 

Leu Asn Thr Gly Trp Leu Ala Gly Leu Phe Tyr Gln His Lys Phe Asn 
        435                 440                 445 

Ser Ser Gly Cys Pro Glu Arg Leu Ala Ser Cys Arg Arg Leu Thr Asp 
    450                 455                 460 

Phe Ala Gln Gly Trp Gly Pro Ile Ser Tyr Ala Asn Gly Ser Gly Leu 
465                 470                 475                 480 

Asp Glu Arg Pro Tyr Cys Trp His Tyr Pro Pro Arg Pro Cys Gly Ile 
                485                 490                 495 

Val Pro Ala Lys Ser Val Cys Gly Pro Val Tyr Cys Phe Thr Pro Ser 
            500                 505                 510 

Pro Val Val Val Gly Thr Thr Asp Arg Ser Gly Ala Pro Thr Tyr Ser 
        515                 520                 525 

Trp Gly Ala Asn Asp Thr Asp Val Phe Val Leu Asn Asn Thr Arg Pro 
    530                 535                 540 

Pro Leu Gly Asn Trp Phe Gly Cys Thr Trp Met Asn Ser Thr Gly Phe 
545                 550                 555                 560 

Thr Lys Val Cys Gly Ala Pro Pro Cys Val Ile Gly Gly Val Gly Asn 
                565                 570                 575 

Asn Thr Leu Leu Cys Pro Thr Asp Cys Phe Arg Lys Tyr Pro Glu Ala 
            580                 585                 590 

Thr Tyr Ser Arg Cys Gly Ser Gly Pro Arg Ile Thr Pro Arg Cys Met 
        595                 600                 605 

Val Asp Tyr Pro Tyr Arg Leu Trp His Tyr Pro Cys Thr Ile Asn Tyr 
    610                 615                 620 

Thr Ile Phe Lys Val Arg Met Tyr Val Gly Gly Val Glu His Arg Leu 
625                 630                 635                 640 

Glu Ala Ala Cys Asn Trp Thr Arg Gly Glu Arg Cys Asp Leu Glu Asp 
                645                 650                 655 

Arg Asp Arg Ser Glu Leu Ser Pro Leu Leu Leu Ser Thr Thr Gln Trp 
            660                 665                 670 

Gln Val Leu Pro Cys Ser Phe Thr Thr Leu Pro Ala Leu Ser Thr Gly 
        675                 680                 685 

Leu Ile His Leu His Gln Asn Ile Val Asp Val Gln Tyr Leu Tyr Gly 
    690                 695                 700 

Val Gly Ser Ser Ile Ala Ser Trp Ala Ile Lys Trp Glu Tyr Val Val 
705                 710                 715                 720 

Leu Leu Phe Leu Leu Leu Ala Asp Ala Arg Val Cys Ser Cys Leu Trp 
                725                 730                 735 

Met Met Leu Leu Ile Ser Gln Ala Glu Ala Ala Leu Glu Asn Leu Val 
            740                 745                 750 

Ile Leu Asn Ala Ala Ser Leu Ala Gly Thr His Gly Leu Val Ser Phe 
        755                 760                 765 

Leu Val Phe Phe Cys Phe Ala Trp Tyr Leu Lys Gly Arg Trp Val Pro 
    770                 775                 780 

Gly Ala Val Tyr Ala Leu Tyr Gly Met Trp Pro Leu Leu Leu Leu Leu 
785                 790                 795                 800 

Leu Ala Leu Pro Gln Arg Ala Tyr Ala Leu Asp Thr Glu Val Ala Ala 
                805                 810                 815 

Ser Cys Gly Gly Val Val Leu Val Gly Leu Met Ala Leu Thr Leu Ser 
            820                 825                 830 

Pro Tyr Tyr Lys Arg Tyr Ile Ser Trp Cys Met Trp Trp Leu Gln Tyr 
        835                 840                 845 

Phe Leu Thr Arg Val Glu Ala Gln Leu His Val Trp Val Pro Pro Leu 
    850                 855                 860 

Asn Val Arg Gly Gly Arg Asp Ala Val Ile Leu Leu Thr Cys Val Val 
865                 870                 875                 880 

His Pro Ala Leu Val Phe Asp Ile Thr Lys Leu Leu Leu Ala Ile Phe 
                885                 890                 895 

Gly Pro Leu Trp Ile Leu Gln Ala Ser Leu Leu Lys Val Pro Tyr Phe 
            900                 905                 910 

Val Arg Val Gln Gly Leu Leu Arg Ile Cys Ala Leu Ala Arg Lys Ile 
        915                 920                 925 

Ala Gly Gly His Tyr Val Gln Met Ala Ile Ile Lys Leu Gly Ala Leu 
    930                 935                 940 

Thr Gly Thr Cys Val Tyr Asn His Leu Ala Pro Leu Arg Asp Trp Ala 
945                 950                 955                 960 

His Asn Gly Leu Arg Asp Leu Ala Val Ala Val Glu Pro Val Val Phe 
                965                 970                 975 

Ser Arg Met Glu Thr Lys Leu Ile Thr Trp Gly Ala Asp Thr Ala Ala 
            980                 985                 990 

Cys Gly Asp Ile Ile Asn Gly Leu Pro Val Ser Ala Arg Arg Gly Gln 
        995                 1000                1005 

Glu Ile Leu Leu Gly Pro Ala Asp Gly Met Val Ser Lys Gly Trp Arg 
    1010                1015                1020 

Leu Leu Ala Pro Ile Thr Ala Tyr Ala Gln Gln Thr Arg Gly Leu Leu 
1025                1030                1035                1040 

Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu 
                1045                1050                1055 

Gly Glu Val Gln Ile Val Ser Thr Ala Thr Gln Thr Phe Leu Ala Thr 
            1060                1065                1070 

Cys Ile Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Thr Arg 
        1075                1080                1085 

Thr Ile Ala Ser Pro Lys Gly Pro Val Ile Gln Thr Tyr Thr Asn Val 
    1090                1095                1100 

Asp Gln Asp Leu Val Gly Trp Pro Ala Pro Gln Gly Ser Arg Ser Leu 
1105                1110                1115                1120 

Thr Pro Cys Thr Cys Gly Ser Ser Asp Leu Tyr Leu Val Thr Arg His 
                1125                1130                1135 

Ala Asp Val Ile Pro Val Arg Arg Arg Gly Asp Ser Arg Gly Ser Leu 
            1140                1145                1150 

Leu Ser Pro Arg Pro Ile Ser Tyr Leu Lys Gly Ser Ser Gly Gly Pro 
        1155                1160                1165 

Leu Leu Cys Pro Thr Gly His Ala Val Gly Leu Phe Arg Ala Ala Val 
    1170                1175                1180 

Cys Thr Arg Gly Val Ala Lys Ala Val Asp Phe Ile Pro Val Glu Asn 
1185                1190                1195                1200 

Leu Glu Thr Thr Met Arg Ser Pro Val Phe Thr Asp Asn Ser Ser Pro 
                1205                1210                1215 

Pro Ala Val Pro Gln Ser Phe Gln Val Ala His Leu His Ala Pro Thr 
            1220                1225                1230 

Gly Ser Gly Lys Ser Thr Lys Val Pro Ala Ala Tyr Ala Ala Lys Gly 
        1235                1240                1245 

Tyr Lys Val Leu Val Leu Asn Pro Ser Val Ala Ala Thr Leu Gly Phe 
    1250                1255                1260 

Gly Ala Tyr Met Ser Lys Ala His Gly Val Asp Pro Asn Ile Arg Thr 
1265                1270                1275                1280 

Gly Val Arg Thr Ile Thr Thr Gly Ser Pro Ile Thr Tyr Ser Thr Tyr 
                1285                1290                1295 

Gly Lys Phe Leu Ala Asp Ala Gly Cys Ser Gly Gly Ala Tyr Asp Ile 
            1300                1305                1310 

Ile Ile Cys Asp Glu Cys His Ser Thr Asp Ala Thr Ser Ile Ser Gly 
        1315                1320                1325 

Ile Gly Thr Val Leu Asp Gln Ala Glu Thr Ala Gly Ala Arg Leu Val 
    1330                1335                1340 

Val Leu Ala Thr Ala Thr Pro Pro Gly Ser Val Thr Val Ser His Pro 
1345                1350                1355                1360 

Asn Ile Glu Glu Val Ala Leu Ser Thr Thr Gly Glu Ile Pro Phe Tyr 
                1365                1370                1375 

Gly Lys Ala Ile Pro Leu Glu Val Ile Lys Gly Gly Arg His Leu Ile 
            1380                1385                1390 

Phe Cys His Ser Lys Lys Lys Cys Asp Glu Leu Ala Ala Lys Leu Val 
        1395                1400                1405 

Ala Leu Gly Ile Asn Ala Val Ala Tyr Tyr Arg Gly Leu Asp Val Ser 
    1410                1415                1420 

Val Ile Pro Thr Ser Gly Asp Val Val Val Val Ser Thr Asp Ala Leu 
1425                1430                1435                1440 

Met Thr Gly Phe Thr Gly Asp Phe Asp Ser Val Ile Asp Cys Asn Thr 
                1445                1450                1455 

Cys Val Thr Gln Thr Val Asp Phe Ser Leu Asp Pro Thr Phe Thr Ile 
            1460                1465                1470 

Glu Thr Thr Thr Leu Pro Gln Asp Ala Val Ser Arg Thr Gln Arg Arg 
        1475                1480                1485 

Gly Arg Thr Gly Arg Gly Lys Pro Gly Ile Tyr Arg Phe Val Ala Pro 
    1490                1495                1500 

Gly Glu Arg Pro Ser Gly Met Phe Asp Ser Ser Val Leu Cys Glu Cys 
1505                1510                1515                1520 

Tyr Asp Ala Gly Cys Ala Trp Tyr Glu Leu Thr Pro Ala Glu Thr Thr 
                1525                1530                1535 

Val Arg Leu Arg Ala Tyr Met Asn Thr Pro Gly Leu Pro Val Cys Gln 
            1540                1545                1550 

Asp His Leu Gly Phe Trp Glu Gly Val Phe Thr Gly Leu Thr His Ile 
        1555                1560                1565 

Asp Ala His Phe Leu Ser Gln Thr Lys Gln Ser Gly Glu Asn Phe Pro 
    1570                1575                1580 

Tyr Leu Val Ala Tyr Gln Ala Thr Val Cys Ala Arg Ala Gln Ala Pro 
1585                1590                1595                1600 

Pro Pro Ser Trp Asp Gln Met Arg Lys Cys Leu Ile Arg Leu Lys Pro 
                1605                1610                1615 

Thr Leu His Gly Pro Thr Pro Leu Leu Tyr Arg Leu Gly Ala Val Gln 
            1620                1625                1630 

Asn Glu Val Thr Leu Thr His Pro Ile Thr Lys Tyr Ile Met Thr Cys 
        1635                1640                1645 

Met Ser Ala Asp Leu Glu Val Val Thr Ser Thr Trp Val Leu Val Gly 
    1650                1655                1660 

Gly Val Leu Ala Ala Leu Ala Ala Tyr Cys Leu Ser Thr Gly Cys Val 
1665                1670                1675                1680 

Val Ile Val Gly Arg Ile Val Leu Ser Gly Lys Pro Ala Ile Ile Pro 
                1685                1690                1695 

Asp Arg Glu Val Leu Tyr Gln Glu Phe Asp Glu Met Glu Glu Cys Ser 
            1700                1705                1710 

Gln His Leu Pro Tyr Ile Glu Gln Gly Met Met Leu Ala Glu Gln Phe 
        1715                1720                1725 

Lys Gln Lys Ala Leu Gly Leu Leu Gln Thr Ala Ser Arg His Ala Glu 
    1730                1735                1740 

Val Ile Thr Pro Ala Val Gln Thr Asn Trp Gln Lys Leu Glu Val Phe 
1745                1750                1755                1760 

Trp Ala Lys His Met Trp Asn Phe Ile Ser Gly Ile Gln Tyr Leu Ala 
                1765                1770                1775 

Gly Leu Ser Thr Leu Pro Gly Asn Pro Ala Ile Ala Ser Leu Met Ala 
            1780                1785                1790 

Phe Thr Ala Ala Val Thr Ser Pro Leu Thr Thr Gly Gln Thr Leu Leu 
        1795                1800                1805 

Phe Asn Ile Leu Gly Gly Trp Val Ala Ala Gln Leu Ala Ala Pro Gly 
    1810                1815                1820 

Ala Ala Thr Ala Phe Val Gly Ala Gly Leu Ala Gly Ala Ala Leu Asp 
1825                1830                1835                1840 

Ser Val Gly Leu Gly Lys Val Leu Val Asp Ile Leu Ala Gly Tyr Gly 
                1845                1850                1855 

Ala Gly Val Ala Gly Ala Leu Val Ala Phe Lys Ile Met Ser Gly Glu 
            1860                1865                1870 

Val Pro Ser Thr Glu Asp Leu Val Asn Leu Leu Pro Ala Ile Leu Ser 
        1875                1880                1885 

Pro Gly Ala Leu Ala Val Gly Val Val Phe Ala Ser Ile Leu Arg Arg 
    1890                1895                1900 

Arg Val Gly Pro Gly Glu Gly Ala Val Gln Trp Met Asn Arg Leu Ile 
1905                1910                1915                1920 

Ala Phe Ala Ser Arg Gly Asn His Val Ser Pro Thr His Tyr Val Pro 
                1925                1930                1935 

Glu Ser Asp Ala Ala Ala Arg Val Thr Ala Ile Leu Ser Ser Leu Thr 
            1940                1945                1950 

Val Thr Gln Leu Leu Arg Arg Leu His Gln Trp Ile Ser Ser Glu Cys 
        1955                1960                1965 

Thr Thr Pro Cys Ser Gly Ser Trp Leu Arg Asp Ile Trp Asp Trp Ile 
    1970                1975                1980 

Cys Glu Val Leu Ser Asp Phe Lys Thr Trp Leu Lys Ala Lys Leu Met 
1985                1990                1995                2000 

Pro Gln Leu Pro Gly Ile Pro Phe Val Ser Cys Gln Arg Gly Tyr Arg 
                2005                2010                2015 

Gly Val Trp Arg Gly Asp Gly Ile Met His Thr Arg Cys His Cys Gly 
            2020                2025                2030 

Ala Glu Ile Thr Gly His Val Lys Asn Gly Thr Met Arg Ile Val Gly 
        2035                2040                2045 

Pro Arg Thr Cys Lys Asn Met Trp Ser Gly Thr Phe Phe Ile Asn Ala 
    2050                2055                2060 

Tyr Thr Thr Gly Pro Cys Thr Pro Leu Pro Ala Pro Asn Tyr Lys Phe 
2065                2070                2075                2080 

Ala Leu Trp Arg Val Ser Ala Glu Glu Tyr Val Glu Ile Arg Arg Val 
                2085                2090                2095 

Gly Asp Phe His Tyr Val Ser Gly Met Thr Thr Asp Asn Leu Lys Cys 
            2100                2105                2110 

Pro Cys Gln Ile Pro Ser Pro Glu Phe Phe Thr Glu Leu Asp Gly Val 
        2115                2120                2125 

Arg Leu His Arg Phe Ala Pro Pro Cys Lys Pro Leu Leu Arg Glu Glu 
    2130                2135                2140 

Val Ser Phe Arg Val Gly Leu His Glu Tyr Pro Val Gly Ser Gln Leu 
2145                2150                2155                2160 

Pro Cys Glu Pro Glu Pro Asp Val Ala Val Leu Thr Ser Met Leu Thr 
                2165                2170                2175 

Asp Pro Ser His Ile Thr Ala Glu Ala Ala Gly Arg Arg Leu Ala Arg 
            2180                2185                2190 

Gly Ser Pro Pro Ser Met Ala Ser Ser Ser Ala Ser Gln Leu Ser Ala 
        2195                2200                2205 

Pro Ser Leu Lys Ala Thr Cys Thr Ala Asn His Asp Ser Pro Asp Ala 
    2210                2215                2220 

Glu Leu Ile Glu Ala Asn Leu Leu Trp Arg Gln Glu Met Gly Gly Asn 
2225                2230                2235                2240 

Ile Thr Arg Val Glu Ser Glu Asn Lys Val Val Ile Leu Asp Ser Phe 
                2245                2250                2255 

Asp Pro Leu Val Ala Glu Glu Asp Glu Arg Glu Val Ser Val Pro Ala 
            2260                2265                2270 

Glu Ile Leu Arg Lys Ser Arg Arg Phe Ala Pro Ala Leu Pro Val Trp 
        2275                2280                2285 

Ala Arg Pro Asp Tyr Asn Pro Leu Leu Val Glu Thr Trp Lys Lys Pro 
    2290                2295                2300 

Asp Tyr Glu Pro Pro Val Val His Gly Cys Pro Leu Pro Pro Pro Arg 
2305                2310                2315                2320 

Ser Pro Pro Val Pro Pro Pro Arg Lys Lys Arg Thr Val Val Leu Thr 
                2325                2330                2335 

Glu Ser Thr Leu Pro Thr Ala Leu Ala Glu Leu Ala Thr Lys Ser Phe 
            2340                2345                2350 

Gly Ser Ser Ser Thr Ser Gly Ile Thr Gly Asp Asn Thr Thr Thr Ser 
        2355                2360                2365 

Ser Glu Pro Ala Pro Ser Gly Cys Pro Pro Asp Ser Asp Val Glu Ser 
    2370                2375                2380 

Tyr Ser Ser Met Pro Pro Leu Glu Gly Glu Pro Gly Asp Pro Asp Leu 
2385                2390                2395                2400 

Ser Asp Gly Ser Trp Ser Thr Val Ser Ser Gly Ala Asp Thr Glu Asp 
                2405                2410                2415 

Val Val Cys Cys Ser Met Ser Tyr Ser Trp Thr Gly Ala Leu Val Thr 
            2420                2425                2430 

Pro Cys Ala Ala Glu Glu Gln Lys Leu Pro Ile Asn Ala Leu Ser Asn 
        2435                2440                2445 

Ser Leu Leu Arg His His Asn Leu Val Tyr Ser Thr Thr Ser Arg Ser 
    2450                2455                2460 

Ala Cys Gln Arg Lys Lys Lys Val Thr Phe Asp Arg Leu Gln Val Leu 
2465                2470                2475                2480 

Asp Ser His Tyr Gln Asp Val Leu Lys Glu Val Lys Ala Ala Ala Ser 
                2485                2490                2495 

Lys Val Lys Ala Asn Leu Leu Ser Val Glu Glu Ala Cys Ser Leu Ala 
            2500                2505                2510 

Pro Pro His Ser Ala Lys Ser Lys Phe Gly Tyr Gly Ala Lys Asp Val 
        2515                2520                2525 

Arg Cys His Ala Arg Lys Ala Val Ala His Ile Asn Ser Val Trp Lys 
    2530                2535                2540 

Asp Leu Leu Glu Asp Ser Val Thr Pro Ile Asp Thr Thr Ile Met Ala 
2545                2550                2555                2560 

Lys Asn Glu Val Phe Cys Val Gln Pro Glu Lys Gly Gly Arg Lys Pro 
                2565                2570                2575 

Ala Arg Leu Ile Val Phe Pro Asp Leu Gly Val Arg Val Cys Glu Lys 
            2580                2585                2590 

Met Ala Leu Tyr Asp Val Val Ser Lys Leu Pro Leu Ala Val Met Gly 
        2595                2600                2605 

Ser Ser Tyr Gly Phe Gln Tyr Ser Pro Gly Gln Arg Val Glu Phe Leu 
    2610                2615                2620 

Val Gln Ala Trp Lys Ser Lys Lys Thr Pro Met Gly Leu Ser Tyr Asp 
2625                2630                2635                2640 

Thr Arg Cys Phe Asp Ser Thr Val Thr Glu Ser Asp Ile Arg Thr Glu 
                2645                2650                2655 

Glu Ala Ile Tyr Gln Cys Cys Asp Leu Asp Pro Gln Ala Arg Val Ala 
            2660                2665                2670 

Ile Lys Ser Leu Thr Glu Arg Leu Tyr Val Gly Gly Pro Leu Thr Asn 
        2675                2680                2685 

Ser Arg Gly Glu Asn Cys Gly Tyr Arg Arg Cys Arg Ala Ser Arg Val 
    2690                2695                2700 

Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr Arg Tyr Ile Lys Ala Arg 
2705                2710                2715                2720 

Ala Ala Cys Arg Ala Ala Gly Leu Gln Asp Cys Thr Met Leu Val Cys 
                2725                2730                2735 

Gly Asp Asp Leu Val Val Ile Cys Glu Ser Ala Gly Val Gln Glu Asp 
            2740                2745                2750 

Ala Ala Ser Leu Arg Ala Phe Thr Glu Ala Met Thr Arg Tyr Ser Ala 
        2755                2760                2765 

Pro Pro Gly Asp Pro Pro Gln Pro Glu Tyr Asp Leu Glu Leu Ile Thr 
    2770                2775                2780 

Ser Cys Ser Ser Asn Val Ser Val Ala His Asp Gly Ala Gly Lys Arg 
2785                2790                2795                2800 

Val Tyr Tyr Leu Thr Arg Asp Pro Thr Thr Pro Leu Ala Arg Ala Ala 
                2805                2810                2815 

Trp Glu Thr Ala Arg His Thr Pro Val Asn Ser Trp Leu Gly Asn Ile 
            2820                2825                2830 

Ile Met Phe Ala Pro Thr Leu Trp Ala Arg Met Ile Leu Met Thr His 
        2835                2840                2845 

Phe Phe Ser Val Leu Ile Ala Arg Asp Gln Leu Glu Gln Ala Leu Asn 
    2850                2855                2860 

Cys Glu Ile Tyr Gly Ala Cys Tyr Ser Ile Glu Pro Leu Asp Leu Pro 
2865                2870                2875                2880 

Pro Ile Ile Gln Arg Leu His Gly Leu Ser Ala Phe Ser Leu His Ser 
                2885                2890                2895 

Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala Ala Cys Leu Arg Lys Leu 
            2900                2905                2910 

Gly Val Pro Pro Leu Arg Ala Trp Arg His Arg Ala Trp Ser Val Arg 
        2915                2920                2925 

Ala Arg Leu Leu Ala Arg Gly Gly Lys Ala Ala Ile Cys Gly Lys Tyr 
    2930                2935                2940 

Leu Phe Asn Trp Ala Val Arg Thr Lys Leu Lys Leu Thr Pro Ile Thr 
2945                2950                2955                2960 

Ala Ala Gly Arg Leu Asp Leu Ser Gly Trp Phe Thr Ala Gly Tyr Ser 
                2965                2970                2975 

Gly Gly Asp Ile Tyr His Ser Val Ser His Ala Arg Pro Arg Trp Phe 
            2980                2985                2990 

Trp Phe Cys Leu Leu Leu Leu Ala Ala Gly Val Gly Ile Tyr Leu Leu 
        2995                3000                3005 

Pro Asn Arg 
    3010 

 
           
             2  
             182  
             PRT  
             Artificial Sequence  
             
               Hepatitis C virus core protein sequence  
             
           
            2 

Met Ser Thr Asn Pro Lys Pro Gln Arg Lys Thr Lys Arg Asn Thr Asn 
 1               5                  10                  15 

Arg Arg Pro Gln Asp Val Lys Phe Pro Gly Gly Gly Gln Ile Val Gly 
            20                  25                  30 

Gly Val Tyr Leu Leu Pro Arg Arg Gly Pro Arg Leu Gly Val Arg Ala 
        35                  40                  45 

Thr Arg Lys Thr Ser Glu Arg Ser Gln Pro Arg Gly Arg Arg Gln Pro 
    50                  55                  60 

Ile Pro Lys Ala Arg Arg Pro Glu Gly Arg Thr Trp Ala Gln Pro Gly 
65                  70                  75                  80 

Tyr Pro Trp Pro Leu Tyr Gly Asn Glu Gly Cys Gly Trp Ala Gly Trp 
                85                  90                  95 

Leu Leu Ser Pro Arg Gly Ser Arg Pro Ser Trp Gly Pro Thr Asp Pro 
            100                 105                 110 

Arg Arg Arg Ser Arg Asn Leu Gly Lys Val Ile Asp Thr Leu Thr Cys 
        115                 120                 125 

Gly Phe Ala Asp Leu Met Gly Tyr Ile Pro Leu Val Gly Ala Pro Leu 
    130                 135                 140 

Gly Gly Ala Ala Arg Ala Leu Ala His Gly Val Arg Val Leu Glu Asp 
145                 150                 155                 160 

Gly Val Asn Tyr Ala Thr Gly Asn Leu Pro Gly Cys Ser Phe Ser Ile 
                165                 170                 175 

Phe Leu Leu Ala Leu Leu 
            180 

 
           
             3  
             197  
             PRT  
             Artificial Sequence  
             
               Hepatitis C virus E1 protein sequence  
             
           
            3 

Ser Cys Leu Thr Val Pro Ala Ser Ala Tyr Gln Val Arg Asn Ser Ser 
 1               5                  10                  15 

Gly Leu Tyr His Val Thr Asn Asp Cys Pro Asn Ser Ser Val Val Tyr 
            20                  25                  30 

Glu Ala Ala Asp Ala Ile Leu His Thr Pro Gly Cys Val Pro Cys Val 
        35                  40                  45 

Arg Glu Gly Asn Ala Ser Arg Cys Trp Val Ala Val Thr Pro Thr Val 
    50                  55                  60 

Ala Thr Arg Asp Gly Lys Leu Pro Thr Thr Gln Leu Arg Arg His Ile 
65                  70                  75                  80 

Asp Leu Leu Val Gly Ser Ala Thr Leu Cys Ser Ala Leu Tyr Val Gly 
                85                  90                  95 

Asp Leu Cys Gly Ser Val Phe Leu Val Gly Gln Leu Phe Thr Phe Ser 
            100                 105                 110 

Pro Arg His His Trp Thr Thr Gln Asp Cys Asn Cys Ser Ile Tyr Pro 
        115                 120                 125 

Gly His Ile Thr Gly His Arg Met Ala Trp Asn Met Met Met Asn Trp 
    130                 135                 140 

Ser Pro Thr Ala Ala Leu Val Val Ala Gln Leu Leu Arg Ile Pro Gln 
145                 150                 155                 160 

Ala Ile Met Asp Met Ile Ala Gly Ala His Trp Gly Val Leu Ala Gly 
                165                 170                 175 

Ile Lys Tyr Phe Ser Met Val Gly Asn Trp Ala Lys Val Leu Val Val 
            180                 185                 190 

Leu Leu Leu Phe Ala 
        195 

 
           
             4  
             350  
             PRT  
             Artificial Sequence  
             
               Hepatitis C virus E2 protein sequence  
             
           
            4 

Gly Val Asp Ala Glu Thr His Val Thr Gly Gly Asn Ala Gly Arg Thr 
 1               5                  10                  15 

Thr Ala Gly Leu Val Gly Leu Leu Thr Pro Gly Ala Lys Gln Asn Ile 
            20                  25                  30 

Gln Leu Ile Asn Thr Asn Gly Ser Trp His Ile Asn Ser Thr Ala Leu 
        35                  40                  45 

Asn Cys Asn Glu Ser Leu Asn Thr Gly Trp Leu Ala Gly Leu Phe Tyr 
    50                  55                  60 

Gln His Lys Phe Asn Ser Ser Gly Cys Pro Glu Arg Leu Ala Ser Cys 
65                  70                  75                  80 

Arg Arg Leu Thr Asp Phe Ala Gln Gly Trp Gly Pro Ile Ser Tyr Ala 
                85                  90                  95 

Asn Gly Ser Gly Leu Asp Glu Arg Pro Tyr Cys Trp His Tyr Pro Pro 
            100                 105                 110 

Arg Pro Cys Gly Ile Val Pro Ala Lys Ser Val Cys Gly Pro Val Tyr 
        115                 120                 125 

Cys Phe Thr Pro Ser Pro Val Val Val Gly Thr Thr Asp Arg Ser Gly 
    130                 135                 140 

Ala Pro Thr Tyr Ser Trp Gly Ala Asn Asp Thr Asp Val Phe Val Leu 
145                 150                 155                 160 

Asn Asn Thr Arg Pro Pro Leu Gly Asn Trp Phe Gly Cys Thr Trp Met 
                165                 170                 175 

Asn Ser Thr Gly Phe Thr Lys Val Cys Gly Ala Pro Pro Cys Val Ile 
            180                 185                 190 

Gly Gly Val Gly Asn Asn Thr Leu Leu Cys Pro Thr Asp Cys Phe Arg 
        195                 200                 205 

Lys Tyr Pro Glu Ala Thr Tyr Ser Arg Cys Gly Ser Gly Pro Arg Ile 
    210                 215                 220 

Thr Pro Arg Cys Met Val Asp Tyr Pro Tyr Arg Leu Trp His Tyr Pro 
225                 230                 235                 240 

Cys Thr Ile Asn Tyr Thr Ile Phe Lys Val Arg Met Tyr Val Gly Gly 
                245                 250                 255 

Val Glu His Arg Leu Glu Ala Ala Cys Asn Trp Thr Arg Gly Glu Arg 
            260                 265                 270 

Cys Asp Leu Glu Asp Arg Asp Arg Ser Glu Leu Ser Pro Leu Leu Leu 
        275                 280                 285 

Ser Thr Thr Gln Trp Gln Val Leu Pro Cys Ser Phe Thr Thr Leu Pro 
    290                 295                 300 

Ala Leu Ser Thr Gly Leu Ile His Leu His Gln Asn Ile Val Asp Val 
305                 310                 315                 320 

Gln Tyr Leu Tyr Gly Val Gly Ser Ser Ile Ala Ser Trp Ala Ile Lys 
                325                 330                 335 

Trp Glu Tyr Val Val Leu Leu Phe Leu Leu Leu Ala Asp Ala 
            340                 345                 350 

 
           
             5  
             315  
             PRT  
             Artificial Sequence  
             
               Hepatitis C virus NS2 protein sequence  
             
           
            5 

Arg Val Cys Ser Cys Leu Trp Met Met Leu Leu Ile Ser Gln Ala Glu 
 1               5                  10                  15 

Ala Ala Leu Glu Asn Leu Val Ile Leu Asn Ala Ala Ser Leu Ala Gly 
            20                  25                  30 

Thr His Gly Leu Val Ser Phe Leu Val Phe Phe Cys Phe Ala Trp Tyr 
        35                  40                  45 

Leu Lys Gly Arg Trp Val Pro Gly Ala Val Tyr Ala Leu Tyr Gly Met 
    50                  55                  60 

Trp Pro Leu Leu Leu Leu Leu Leu Ala Leu Pro Gln Arg Ala Tyr Ala 
65                  70                  75                  80 

Leu Asp Thr Glu Val Ala Ala Ser Cys Gly Gly Val Val Leu Val Gly 
                85                  90                  95 

Leu Met Ala Leu Thr Leu Ser Pro Tyr Tyr Lys Arg Tyr Ile Ser Trp 
            100                 105                 110 

Cys Met Trp Trp Leu Gln Tyr Phe Leu Thr Arg Val Glu Ala Gln Leu 
        115                 120                 125 

His Val Trp Val Pro Pro Leu Asn Val Arg Gly Gly Arg Asp Ala Val 
    130                 135                 140 

Ile Leu Leu Thr Cys Val Val His Pro Ala Leu Val Phe Asp Ile Thr 
145                 150                 155                 160 

Lys Leu Leu Leu Ala Ile Phe Gly Pro Leu Trp Ile Leu Gln Ala Ser 
                165                 170                 175 

Leu Leu Lys Val Pro Tyr Phe Val Arg Val Gln Gly Leu Leu Arg Ile 
            180                 185                 190 

Cys Ala Leu Ala Arg Lys Ile Ala Gly Gly His Tyr Val Gln Met Ala 
        195                 200                 205 

Ile Ile Lys Leu Gly Ala Leu Thr Gly Thr Cys Val Tyr Asn His Leu 
    210                 215                 220 

Ala Pro Leu Arg Asp Trp Ala His Asn Gly Leu Arg Asp Leu Ala Val 
225                 230                 235                 240 

Ala Val Glu Pro Val Val Phe Ser Arg Met Glu Thr Lys Leu Ile Thr 
                245                 250                 255 

Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Asn Gly Leu Pro 
            260                 265                 270 

Val Ser Ala Arg Arg Gly Gln Glu Ile Leu Leu Gly Pro Ala Asp Gly 
        275                 280                 285 

Met Val Ser Lys Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala Tyr Ala 
    290                 295                 300 

Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile 
305                 310                 315 

 
           
             6  
             613  
             PRT  
             Artificial Sequence  
             
               Hepatitis C virus NS3 protein sequence  
             
           
            6 

Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu Gly Glu Val Gln 
 1               5                  10                  15 

Ile Val Ser Thr Ala Thr Gln Thr Phe Leu Ala Thr Cys Ile Asn Gly 
            20                  25                  30 

Val Cys Trp Thr Val Tyr His Gly Ala Gly Thr Arg Thr Ile Ala Ser 
        35                  40                  45 

Pro Lys Gly Pro Val Ile Gln Thr Tyr Thr Asn Val Asp Gln Asp Leu 
    50                  55                  60 

Val Gly Trp Pro Ala Pro Gln Gly Ser Arg Ser Leu Thr Pro Cys Thr 
65                  70                  75                  80 

Cys Gly Ser Ser Asp Leu Tyr Leu Val Thr Arg His Ala Asp Val Ile 
                85                  90                  95 

Pro Val Arg Arg Arg Gly Asp Ser Arg Gly Ser Leu Leu Ser Pro Arg 
            100                 105                 110 

Pro Ile Ser Tyr Leu Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro 
        115                 120                 125 

Thr Gly His Ala Val Gly Leu Phe Arg Ala Ala Val Cys Thr Arg Gly 
    130                 135                 140 

Val Ala Lys Ala Val Asp Phe Ile Pro Val Glu Asn Leu Glu Thr Thr 
145                 150                 155                 160 

Met Arg Ser Pro Val Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro 
                165                 170                 175 

Gln Ser Phe Gln Val Ala His Leu His Ala Pro Thr Gly Ser Gly Lys 
            180                 185                 190 

Ser Thr Lys Val Pro Ala Ala Tyr Ala Ala Lys Gly Tyr Lys Val Leu 
        195                 200                 205 

Val Leu Asn Pro Ser Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met 
    210                 215                 220 

Ser Lys Ala His Gly Val Asp Pro Asn Ile Arg Thr Gly Val Arg Thr 
225                 230                 235                 240 

Ile Thr Thr Gly Ser Pro Ile Thr Tyr Ser Thr Tyr Gly Lys Phe Leu 
                245                 250                 255 

Ala Asp Ala Gly Cys Ser Gly Gly Ala Tyr Asp Ile Ile Ile Cys Asp 
            260                 265                 270 

Glu Cys His Ser Thr Asp Ala Thr Ser Ile Ser Gly Ile Gly Thr Val 
        275                 280                 285 

Leu Asp Gln Ala Glu Thr Ala Gly Ala Arg Leu Val Val Leu Ala Thr 
    290                 295                 300 

Ala Thr Pro Pro Gly Ser Val Thr Val Ser His Pro Asn Ile Glu Glu 
305                 310                 315                 320 

Val Ala Leu Ser Thr Thr Gly Glu Ile Pro Phe Tyr Gly Lys Ala Ile 
                325                 330                 335 

Pro Leu Glu Val Ile Lys Gly Gly Arg His Leu Ile Phe Cys His Ser 
            340                 345                 350 

Lys Lys Lys Cys Asp Glu Leu Ala Ala Lys Leu Val Ala Leu Gly Ile 
        355                 360                 365 

Asn Ala Val Ala Tyr Tyr Arg Gly Leu Asp Val Ser Val Ile Pro Thr 
    370                 375                 380 

Ser Gly Asp Val Val Val Val Ser Thr Asp Ala Leu Met Thr Gly Phe 
385                 390                 395                 400 

Thr Gly Asp Phe Asp Ser Val Ile Asp Cys Asn Thr Cys Val Thr Gln 
                405                 410                 415 

Thr Val Asp Phe Ser Leu Asp Pro Thr Phe Thr Ile Glu Thr Thr Thr 
            420                 425                 430 

Leu Pro Gln Asp Ala Val Ser Arg Thr Gln Arg Arg Gly Arg Thr Gly 
        435                 440                 445 

Arg Gly Lys Pro Gly Ile Tyr Arg Phe Val Ala Pro Gly Glu Arg Pro 
    450                 455                 460 

Ser Gly Met Phe Asp Ser Ser Val Leu Cys Glu Cys Tyr Asp Ala Gly 
465                 470                 475                 480 

Cys Ala Trp Tyr Glu Leu Thr Pro Ala Glu Thr Thr Val Arg Leu Arg 
                485                 490                 495 

Ala Tyr Met Asn Thr Pro Gly Leu Pro Val Cys Gln Asp His Leu Gly 
            500                 505                 510 

Phe Trp Glu Gly Val Phe Thr Gly Leu Thr His Ile Asp Ala His Phe 
        515                 520                 525 

Leu Ser Gln Thr Lys Gln Ser Gly Glu Asn Phe Pro Tyr Leu Val Ala 
    530                 535                 540 

Tyr Gln Ala Thr Val Cys Ala Arg Ala Gln Ala Pro Pro Pro Ser Trp 
545                 550                 555                 560 

Asp Gln Met Arg Lys Cys Leu Ile Arg Leu Lys Pro Thr Leu His Gly 
                565                 570                 575 

Pro Thr Pro Leu Leu Tyr Arg Leu Gly Ala Val Gln Asn Glu Val Thr 
            580                 585                 590 

Leu Thr His Pro Ile Thr Lys Tyr Ile Met Thr Cys Met Ser Ala Asp 
        595                 600                 605 

Leu Glu Val Val Thr 
    610 

 
           
             7  
             54  
             PRT  
             Artificial Sequence  
             
               Hepatitis C virus NS4A protein sequence  
             
           
            7 

Ser Thr Trp Val Leu Val Gly Gly Val Leu Ala Ala Leu Ala Ala Tyr 
 1               5                  10                  15 

Cys Leu Ser Thr Gly Cys Val Val Ile Val Gly Arg Ile Val Leu Ser 
            20                  25                  30 

Gly Lys Pro Ala Ile Ile Pro Asp Arg Glu Val Leu Tyr Gln Glu Phe 
        35                  40                  45 

Asp Glu Met Glu Glu Cys 
    50 

 
           
             8  
             260  
             PRT  
             Artificial Sequence  
             
               Hepatitis C virus NS4B protein sequence  
             
           
            8 

Ser Gln His Leu Pro Tyr Ile Glu Gln Gly Met Met Leu Ala Glu Gln 
 1               5                  10                  15 

Phe Lys Gln Lys Ala Leu Gly Leu Leu Gln Thr Ala Ser Arg His Ala 
            20                  25                  30 

Glu Val Ile Thr Pro Ala Val Gln Thr Asn Trp Gln Lys Leu Glu Val 
        35                  40                  45 

Phe Trp Ala Lys His Met Trp Asn Phe Ile Ser Gly Ile Gln Tyr Leu 
    50                  55                  60 

Ala Gly Leu Ser Thr Leu Pro Gly Asn Pro Ala Ile Ala Ser Leu Met 
65                  70                  75                  80 

Ala Phe Thr Ala Ala Val Thr Ser Pro Leu Thr Thr Gly Gln Thr Leu 
                85                  90                  95 

Leu Phe Asn Ile Leu Gly Gly Trp Val Ala Ala Gln Leu Ala Ala Pro 
            100                 105                 110 

Gly Ala Ala Thr Ala Phe Val Gly Ala Gly Leu Ala Gly Ala Ala Leu 
        115                 120                 125 

Asp Ser Val Gly Leu Gly Lys Val Leu Val Asp Ile Leu Ala Gly Tyr 
    130                 135                 140 

Gly Ala Gly Val Ala Gly Ala Leu Val Ala Phe Lys Ile Met Ser Gly 
145                 150                 155                 160 

Glu Val Pro Ser Thr Glu Asp Leu Val Asn Leu Leu Pro Ala Ile Leu 
                165                 170                 175 

Ser Pro Gly Ala Leu Ala Val Gly Val Val Phe Ala Ser Ile Leu Arg 
            180                 185                 190 

Arg Arg Val Gly Pro Gly Glu Gly Ala Val Gln Trp Met Asn Arg Leu 
        195                 200                 205 

Ile Ala Phe Ala Ser Arg Gly Asn His Val Ser Pro Thr His Tyr Val 
    210                 215                 220 

Pro Glu Ser Asp Ala Ala Ala Arg Val Thr Ala Ile Leu Ser Ser Leu 
225                 230                 235                 240 

Thr Val Thr Gln Leu Leu Arg Arg Leu His Gln Trp Ile Ser Ser Glu 
                245                 250                 255 

Cys Thr Thr Pro 
            260 

 
           
             9  
             1040  
             PRT  
             Artificial Sequence  
             
               Hepatitis C virus NS5A/B protein sequence  
             
           
            9 

Cys Ser Gly Ser Trp Leu Arg Asp Ile Trp Asp Trp Ile Cys Glu Val 
 1               5                  10                  15 

Leu Ser Asp Phe Lys Thr Trp Leu Lys Ala Lys Leu Met Pro Gln Leu 
            20                  25                  30 

Pro Gly Ile Pro Phe Val Ser Cys Gln Arg Gly Tyr Arg Gly Val Trp 
        35                  40                  45 

Arg Gly Asp Gly Ile Met His Thr Arg Cys His Cys Gly Ala Glu Ile 
    50                  55                  60 

Thr Gly His Val Lys Asn Gly Thr Met Arg Ile Val Gly Pro Arg Thr 
65                  70                  75                  80 

Cys Lys Asn Met Trp Ser Gly Thr Phe Phe Ile Asn Ala Tyr Thr Thr 
                85                  90                  95 

Gly Pro Cys Thr Pro Leu Pro Ala Pro Asn Tyr Lys Phe Ala Leu Trp 
            100                 105                 110 

Arg Val Ser Ala Glu Glu Tyr Val Glu Ile Arg Arg Val Gly Asp Phe 
        115                 120                 125 

His Tyr Val Ser Gly Met Thr Thr Asp Asn Leu Lys Cys Pro Cys Gln 
    130                 135                 140 

Ile Pro Ser Pro Glu Phe Phe Thr Glu Leu Asp Gly Val Arg Leu His 
145                 150                 155                 160 

Arg Phe Ala Pro Pro Cys Lys Pro Leu Leu Arg Glu Glu Val Ser Phe 
                165                 170                 175 

Arg Val Gly Leu His Glu Tyr Pro Val Gly Ser Gln Leu Pro Cys Glu 
            180                 185                 190 

Pro Glu Pro Asp Val Ala Val Leu Thr Ser Met Leu Thr Asp Pro Ser 
        195                 200                 205 

His Ile Thr Ala Glu Ala Ala Gly Arg Arg Leu Ala Arg Gly Ser Pro 
    210                 215                 220 

Pro Ser Met Ala Ser Ser Ser Ala Ser Gln Leu Ser Ala Pro Ser Leu 
225                 230                 235                 240 

Lys Ala Thr Cys Thr Ala Asn His Asp Ser Pro Asp Ala Glu Leu Ile 
                245                 250                 255 

Glu Ala Asn Leu Leu Trp Arg Gln Glu Met Gly Gly Asn Ile Thr Arg 
            260                 265                 270 

Val Glu Ser Glu Asn Lys Val Val Ile Leu Asp Ser Phe Asp Pro Leu 
        275                 280                 285 

Val Ala Glu Glu Asp Glu Arg Glu Val Ser Val Pro Ala Glu Ile Leu 
    290                 295                 300 

Arg Lys Ser Arg Arg Phe Ala Pro Ala Leu Pro Val Trp Ala Arg Pro 
305                 310                 315                 320 

Asp Tyr Asn Pro Leu Leu Val Glu Thr Trp Lys Lys Pro Asp Tyr Glu 
                325                 330                 335 

Pro Pro Val Val His Gly Cys Pro Leu Pro Pro Pro Arg Ser Pro Pro 
            340                 345                 350 

Val Pro Pro Pro Arg Lys Lys Arg Thr Val Val Leu Thr Glu Ser Thr 
        355                 360                 365 

Leu Pro Thr Ala Leu Ala Glu Leu Ala Thr Lys Ser Phe Gly Ser Ser 
    370                 375                 380 

Ser Thr Ser Gly Ile Thr Gly Asp Asn Thr Thr Thr Ser Ser Glu Pro 
385                 390                 395                 400 

Ala Pro Ser Gly Cys Pro Pro Asp Ser Asp Val Glu Ser Tyr Ser Ser 
                405                 410                 415 

Met Pro Pro Leu Glu Gly Glu Pro Gly Asp Pro Asp Leu Ser Asp Gly 
            420                 425                 430 

Ser Trp Ser Thr Val Ser Ser Gly Ala Asp Thr Glu Asp Val Val Cys 
        435                 440                 445 

Cys Ser Met Ser Tyr Ser Trp Thr Gly Ala Leu Val Thr Pro Cys Ala 
    450                 455                 460 

Ala Glu Glu Gln Lys Leu Pro Ile Asn Ala Leu Ser Asn Ser Leu Leu 
465                 470                 475                 480 

Arg His His Asn Leu Val Tyr Ser Thr Thr Ser Arg Ser Ala Cys Gln 
                485                 490                 495 

Arg Lys Lys Lys Val Thr Phe Asp Arg Leu Gln Val Leu Asp Ser His 
            500                 505                 510 

Tyr Gln Asp Val Leu Lys Glu Val Lys Ala Ala Ala Ser Lys Val Lys 
        515                 520                 525 

Ala Asn Leu Leu Ser Val Glu Glu Ala Cys Ser Leu Ala Pro Pro His 
    530                 535                 540 

Ser Ala Lys Ser Lys Phe Gly Tyr Gly Ala Lys Asp Val Arg Cys His 
545                 550                 555                 560 

Ala Arg Lys Ala Val Ala His Ile Asn Ser Val Trp Lys Asp Leu Leu 
                565                 570                 575 

Glu Asp Ser Val Thr Pro Ile Asp Thr Thr Ile Met Ala Lys Asn Glu 
            580                 585                 590 

Val Phe Cys Val Gln Pro Glu Lys Gly Gly Arg Lys Pro Ala Arg Leu 
        595                 600                 605 

Ile Val Phe Pro Asp Leu Gly Val Arg Val Cys Glu Lys Met Ala Leu 
    610                 615                 620 

Tyr Asp Val Val Ser Lys Leu Pro Leu Ala Val Met Gly Ser Ser Tyr 
625                 630                 635                 640 

Gly Phe Gln Tyr Ser Pro Gly Gln Arg Val Glu Phe Leu Val Gln Ala 
                645                 650                 655 

Trp Lys Ser Lys Lys Thr Pro Met Gly Leu Ser Tyr Asp Thr Arg Cys 
            660                 665                 670 

Phe Asp Ser Thr Val Thr Glu Ser Asp Ile Arg Thr Glu Glu Ala Ile 
        675                 680                 685 

Tyr Gln Cys Cys Asp Leu Asp Pro Gln Ala Arg Val Ala Ile Lys Ser 
    690                 695                 700 

Leu Thr Glu Arg Leu Tyr Val Gly Gly Pro Leu Thr Asn Ser Arg Gly 
705                 710                 715                 720 

Glu Asn Cys Gly Tyr Arg Arg Cys Arg Ala Ser Arg Val Leu Thr Thr 
                725                 730                 735 

Ser Cys Gly Asn Thr Leu Thr Arg Tyr Ile Lys Ala Arg Ala Ala Cys 
            740                 745                 750 

Arg Ala Ala Gly Leu Gln Asp Cys Thr Met Leu Val Cys Gly Asp Asp 
        755                 760                 765 

Leu Val Val Ile Cys Glu Ser Ala Gly Val Gln Glu Asp Ala Ala Ser 
    770                 775                 780 

Leu Arg Ala Phe Thr Glu Ala Met Thr Arg Tyr Ser Ala Pro Pro Gly 
785                 790                 795                 800 

Asp Pro Pro Gln Pro Glu Tyr Asp Leu Glu Leu Ile Thr Ser Cys Ser 
                805                 810                 815 

Ser Asn Val Ser Val Ala His Asp Gly Ala Gly Lys Arg Val Tyr Tyr 
            820                 825                 830 

Leu Thr Arg Asp Pro Thr Thr Pro Leu Ala Arg Ala Ala Trp Glu Thr 
        835                 840                 845 

Ala Arg His Thr Pro Val Asn Ser Trp Leu Gly Asn Ile Ile Met Phe 
    850                 855                 860 

Ala Pro Thr Leu Trp Ala Arg Met Ile Leu Met Thr His Phe Phe Ser 
865                 870                 875                 880 

Val Leu Ile Ala Arg Asp Gln Leu Glu Gln Ala Leu Asn Cys Glu Ile 
                885                 890                 895 

Tyr Gly Ala Cys Tyr Ser Ile Glu Pro Leu Asp Leu Pro Pro Ile Ile 
            900                 905                 910 

Gln Arg Leu His Gly Leu Ser Ala Phe Ser Leu His Ser Tyr Ser Pro 
        915                 920                 925 

Gly Glu Ile Asn Arg Val Ala Ala Cys Leu Arg Lys Leu Gly Val Pro 
    930                 935                 940 

Pro Leu Arg Ala Trp Arg His Arg Ala Trp Ser Val Arg Ala Arg Leu 
945                 950                 955                 960 

Leu Ala Arg Gly Gly Lys Ala Ala Ile Cys Gly Lys Tyr Leu Phe Asn 
                965                 970                 975 

Trp Ala Val Arg Thr Lys Leu Lys Leu Thr Pro Ile Thr Ala Ala Gly 
            980                 985                 990 

Arg Leu Asp Leu Ser Gly Trp Phe Thr Ala Gly Tyr Ser Gly Gly Asp 
        995                 1000                1005 

Ile Tyr His Ser Val Ser His Ala Arg Pro Arg Trp Phe Trp Phe Cys 
    1010                1015                1020 

Leu Leu Leu Leu Ala Ala Gly Val Gly Ile Tyr Leu Leu Pro Asn Arg 
1025                1030                1035                1040 

 
           
             10  
             226  
             PRT  
             Artificial Sequence  
             
               Hepatitis B virus S antigen (HBsAg) sequence  
             
           
            10 

Met Glu Asn Ile Thr Ser Gly Phe Leu Gly Pro Leu Leu Val Leu Gln 
 1               5                  10                  15 

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

Asp Ser Trp Trp Thr Ser Leu Asn Phe Leu Gly Gly Thr Thr Val Cys 
        35                  40                  45 

Leu Gly Gln Asn Ser Gln Ser Pro Thr Ser Asn His Ser Pro Thr Ser 
    50                  55                  60 

Cys Pro Pro Thr Cys Pro Gly Tyr Arg Trp Met Cys Leu Arg Arg Phe 
65                  70                  75                  80 

Ile Ile Phe Leu Phe Ile Leu Leu Leu Cys Leu Ile Phe Leu Leu Val 
                85                  90                  95 

Leu Leu Asp Tyr Gln Gly Met Leu Pro Val Cys Pro Leu Ile Pro Gly 
            100                 105                 110 

Ser Ser Thr Thr Ser Thr Gly Pro Cys Arg Thr Cys Met Thr Thr Ala 
        115                 120                 125 

Gln Gly Thr Ser Met Tyr Pro Ser Cys Cys Cys Thr Lys Pro Ser Asp 
    130                 135                 140 

Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser Trp Ala Phe Gly Lys 
145                 150                 155                 160 

Phe Leu Trp Glu Trp Ala Ser Ala Arg Phe Ser Trp Leu Ser Leu Leu 
                165                 170                 175 

Val Pro Phe Val Gln Trp Phe Val Gly Leu Ser Pro Thr Val Trp Leu 
            180                 185                 190 

Ser Val Ile Trp Met Met Trp Tyr Trp Gly Pro Ser Leu Tyr Ser Ile 
        195                 200                 205 

Leu Ser Pro Phe Leu Pro Leu Leu Pro Ile Phe Phe Cys Leu Trp Val 
    210                 215                 220 

Tyr Ile 
225 

 
           
             11  
             212  
             PRT  
             Artificial Sequence  
             
               Hepatitis B virus C antigen and e antigen 
      (HBcAg/HBeAg) sequence  
             
           
            11 

Met Gln Leu Phe His Leu Cys Leu Ile Ile Ser Cys Ser Cys Pro Thr 
 1               5                  10                  15 

Val Gln Ala Ser Lys Leu Cys Leu Gly Trp Leu Trp Gly Met Asp Ile 
            20                  25                  30 

Asp Pro Tyr Lys Glu Phe Gly Ala Thr Val Glu Leu Leu Ser Phe Leu 
        35                  40                  45 

Pro Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp Thr Ala Ser 
    50                  55                  60 

Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys Ser Pro His 
65                  70                  75                  80 

His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu Leu Met Thr 
                85                  90                  95 

Leu Ala Thr Trp Val Gly Val Asn Leu Glu Asp Pro Ala Ser Arg Asp 
            100                 105                 110 

Leu Val Val Ser Tyr Val Asn Thr Asn Met Gly Leu Lys Phe Arg Gln 
        115                 120                 125 

Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg Glu Thr Val 
    130                 135                 140 

Ile Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr Pro Pro Ala 
145                 150                 155                 160 

Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu Thr Thr 
                165                 170                 175 

Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr Pro Ser Pro 
            180                 185                 190 

Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser Gln Ser Arg 
        195                 200                 205 

Glu Ser Gln Cys 
    210 

 
           
             12  
             2227  
             PRT  
             Artificial Sequence  
             
               Hepatitis A virus sequence  
             
           
            12 

Met Asn Met Ser Lys Gln Gly Ile Phe Gln Thr Val Gly Ser Gly Leu 
 1               5                  10                  15 

Asp His Ile Leu Ser Leu Ala Asp Ile Glu Glu Glu Gln Met Ile Gln 
            20                  25                  30 

Ser Val Asp Arg Thr Ala Val Thr Gly Ala Ser Tyr Phe Thr Ser Val 
        35                  40                  45 

Asp Gln Ser Ser Val His Thr Ala Glu Val Gly Ser His Gln Ile Glu 
    50                  55                  60 

Pro Leu Lys Thr Ser Val Asp Lys Pro Gly Ser Lys Lys Thr Gln Gly 
65                  70                  75                  80 

Glu Lys Phe Phe Leu Ile His Ser Ala Asp Trp Leu Thr Thr His Ala 
                85                  90                  95 

Leu Phe His Glu Val Ala Lys Leu Asp Val Val Lys Leu Leu Tyr Asn 
            100                 105                 110 

Glu Gln Phe Ala Val Gln Gly Leu Leu Arg Tyr His Thr Tyr Ala Arg 
        115                 120                 125 

Phe Gly Ile Glu Ile Gln Val Gln Ile Asn Pro Thr Pro Phe Gln Gln 
    130                 135                 140 

Gly Gly Leu Ile Cys Ala Met Val Pro Gly Asp Gln Ser Tyr Gly Ser 
145                 150                 155                 160 

Ile Ala Ser Leu Thr Val Tyr Pro His Gly Leu Leu Asn Cys Asn Ile 
                165                 170                 175 

Asn Asn Val Val Arg Ile Lys Val Pro Phe Ile Tyr Thr Arg Gly Ala 
            180                 185                 190 

Tyr His Phe Lys Asp Pro Gln Tyr Pro Val Trp Glu Leu Thr Ile Arg 
        195                 200                 205 

Val Trp Ser Glu Leu Asn Ile Gly Thr Gly Thr Ser Ala Tyr Thr Ser 
    210                 215                 220 

Leu Asn Val Leu Ala Arg Phe Thr Asp Leu Glu Leu His Gly Leu Thr 
225                 230                 235                 240 

Pro Leu Ser Thr Gln Met Met Arg Asn Glu Phe Arg Val Ser Thr Thr 
                245                 250                 255 

Glu Asn Val Val Asn Leu Ser Asn Tyr Glu Asp Ala Arg Ala Lys Met 
            260                 265                 270 

Ser Phe Ala Leu Asp Gln Glu Asp Trp Lys Ser Asp Pro Ser Gln Gly 
        275                 280                 285 

Gly Gly Ile Lys Ile Thr His Phe Thr Thr Trp Thr Ser Ile Pro Thr 
    290                 295                 300 

Leu Ala Ala Gln Phe Pro Phe Asn Ala Ser Asp Ser Val Gly Gln Gln 
305                 310                 315                 320 

Ile Lys Val Ile Pro Val Asp Pro Tyr Phe Phe Gln Met Thr Asn Thr 
                325                 330                 335 

Asn Pro Asp Gln Lys Cys Ile Thr Ala Leu Ala Ser Ile Cys Gln Met 
            340                 345                 350 

Phe Cys Phe Trp Arg Gly Asp Leu Val Phe Asp Phe Gln Val Phe Pro 
        355                 360                 365 

Thr Lys Tyr His Ser Gly Arg Leu Leu Phe Cys Phe Val Pro Gly Asn 
    370                 375                 380 

Glu Leu Ile Asp Val Thr Gly Ile Thr Leu Lys Gln Ala Thr Thr Ala 
385                 390                 395                 400 

Pro Cys Ala Val Met Asp Ile Thr Gly Val Gln Ser Thr Leu Arg Phe 
                405                 410                 415 

Arg Val Pro Trp Ile Ser Asp Thr Pro Tyr Arg Val Asn Arg Tyr Thr 
            420                 425                 430 

Lys Ser Ala His Gln Lys Gly Glu Tyr Thr Ala Ile Gly Lys Leu Ile 
        435                 440                 445 

Val Tyr Cys Tyr Asn Arg Leu Thr Ser Pro Ser Asn Val Ala Ser His 
    450                 455                 460 

Val Arg Val Asn Val Tyr Leu Ser Ala Ile Asn Leu Glu Cys Phe Ala 
465                 470                 475                 480 

Pro Leu Tyr His Ala Met Asp Val Thr Thr Gln Val Gly Asp Asp Ser 
                485                 490                 495 

Gly Gly Phe Ser Thr Thr Val Ser Thr Glu Gln Asn Val Pro Asp Pro 
            500                 505                 510 

Gln Val Gly Ile Thr Thr Met Arg Asp Leu Lys Gly Lys Ala Asn Arg 
        515                 520                 525 

Gly Lys Met Asp Val Ser Gly Val Gln Ala Pro Arg Gly Ser Tyr Gln 
    530                 535                 540 

Gln Gln Leu Asn Asp Pro Val Leu Ala Lys Lys Val Pro Glu Thr Phe 
545                 550                 555                 560 

Pro Glu Leu Lys Pro Gly Glu Ser Arg His Thr Ser Asp His Met Ser 
                565                 570                 575 

Ile Tyr Lys Phe Met Gly Arg Ser His Phe Leu Cys Thr Phe Thr Phe 
            580                 585                 590 

Asn Ser Asn Asn Lys Glu Tyr Thr Phe Pro Ile Thr Leu Ser Ser Thr 
        595                 600                 605 

Ser Asn Pro Pro His Gly Leu Pro Ser Thr Leu Arg Trp Phe Phe Asn 
    610                 615                 620 

Leu Phe Gln Leu Tyr Arg Gly Pro Leu Asp Leu Thr Ile Ile Ile Thr 
625                 630                 635                 640 

Gly Ala Thr Asp Val Asp Gly Met Ala Trp Phe Thr Pro Val Gly Leu 
                645                 650                 655 

Ala Val Asp Pro Trp Val Glu Lys Glu Ser Ala Leu Ser Ile Asp Tyr 
            660                 665                 670 

Lys Thr Ala Leu Gly Ala Val Arg Phe Asn Thr Arg Arg Thr Gly Asn 
        675                 680                 685 

Ile Gln Ile Arg Leu Pro Trp Tyr Ser Tyr Leu Tyr Ala Val Ser Gly 
    690                 695                 700 

Ala Leu Asp Gly Leu Gly Asp Lys Thr Asp Ser Thr Phe Gly Leu Phe 
705                 710                 715                 720 

Leu Phe Glu Ile Ala Asn Tyr Asn His Ser Asp Glu Tyr Leu Ser Phe 
                725                 730                 735 

Ser Cys Tyr Leu Ser Val Thr Glu Gln Ser Glu Phe Tyr Phe Pro Arg 
            740                 745                 750 

Ala Pro Leu Asn Ser Asn Ala Met Leu Ser Thr Glu Ser Met Met Ser 
        755                 760                 765 

Arg Ile Ala Ala Gly Asp Leu Glu Ser Ser Val Asp Asp Pro Arg Ser 
    770                 775                 780 

Glu Glu Asp Arg Arg Phe Glu Ser His Ile Glu Cys Arg Lys Pro Tyr 
785                 790                 795                 800 

Lys Glu Leu Arg Leu Glu Val Gly Lys Gln Arg Leu Lys Tyr Ala Gln 
                805                 810                 815 

Glu Glu Leu Ser Asn Glu Val Leu Pro Pro Pro Arg Lys Met Lys Gly 
            820                 825                 830 

Leu Phe Ser Gln Ala Lys Ile Ser Leu Phe Tyr Thr Glu Glu His Glu 
        835                 840                 845 

Ile Met Lys Phe Ser Trp Arg Gly Val Thr Ala Asp Thr Arg Ala Leu 
    850                 855                 860 

Arg Arg Phe Gly Phe Ser Leu Ala Ala Gly Arg Ser Val Trp Thr Leu 
865                 870                 875                 880 

Glu Met Asp Ala Gly Val Leu Thr Gly Arg Leu Ile Arg Leu Asn Asp 
                885                 890                 895 

Glu Lys Trp Thr Glu Met Lys Asp Asp Lys Ile Val Ser Leu Ile Glu 
            900                 905                 910 

Lys Phe Thr Ser Asn Lys Tyr Trp Ser Lys Val Asn Phe Pro His Gly 
        915                 920                 925 

Met Leu Asp Leu Glu Glu Ile Ala Ala Asn Ser Lys Asp Phe Pro Asn 
    930                 935                 940 

Met Ser Glu Thr Asp Leu Cys Phe Leu Leu His Trp Leu Asn Pro Lys 
945                 950                 955                 960 

Lys Ile Asn Leu Ala Asp Arg Met Leu Gly Leu Ser Gly Val Gln Glu 
                965                 970                 975 

Ile Lys Glu Gln Gly Val Gly Leu Ile Ala Glu Cys Arg Thr Phe Leu 
            980                 985                 990 

Asp Ser Ile Ala Gly Thr Leu Lys Ser Met Met Phe Gly Phe His His 
        995                 1000                1005 

Ser Val Thr Val Glu Ile Ile Asn Thr Val Leu Cys Phe Val Lys Ser 
    1010                1015                1020 

Gly Ile Leu Leu Tyr Val Ile Gln Gln Leu Asn Gln Asp Glu His Ser 
1025                1030                1035                1040 

His Ile Ile Gly Leu Leu Arg Val Met Asn Tyr Ala Asp Ile Gly Cys 
                1045                1050                1055 

Ser Val Ile Ser Cys Gly Lys Val Phe Ser Lys Met Leu Glu Thr Val 
            1060                1065                1070 

Phe Asn Trp Gln Met Asp Ser Arg Met Met Glu Leu Arg Thr Gln Ser 
        1075                1080                1085 

Phe Ser Asn Trp Leu Arg Asp Ile Cys Ser Gly Ile Thr Ile Phe Lys 
    1090                1095                1100 

Ser Phe Lys Asp Ala Ile Tyr Trp Leu Tyr Thr Lys Leu Lys Asp Phe 
1105                1110                1115                1120 

Tyr Glu Val Asn Tyr Gly Lys Lys Lys Asp Ile Leu Asn Ile Leu Lys 
                1125                1130                1135 

Asp Asn Gln Gln Lys Ile Glu Lys Ala Ile Glu Glu Ala Asp Asn Phe 
            1140                1145                1150 

Cys Ile Leu Gln Ile Gln Asp Val Glu Lys Phe Asp Gln Tyr Gln Lys 
        1155                1160                1165 

Gly Val Asp Leu Ile Gln Lys Leu Arg Thr Val His Ser Met Ala Gln 
    1170                1175                1180 

Val Asp Pro Asn Leu Gly Val His Leu Ser Pro Leu Arg Asp Cys Ile 
1185                1190                1195                1200 

Ala Arg Val His Gln Lys Leu Lys Asn Leu Gly Ser Ile Asn Gln Ala 
                1205                1210                1215 

Met Val Thr Arg Cys Glu Pro Val Val Cys Tyr Leu Tyr Gly Lys Arg 
            1220                1225                1230 

Gly Gly Gly Lys Ser Leu Thr Ser Ile Ala Leu Ala Thr Lys Ile Cys 
        1235                1240                1245 

Lys His Tyr Gly Val Glu Pro Glu Lys Asn Ile Tyr Thr Lys Pro Val 
    1250                1255                1260 

Ala Ser Asp Tyr Trp Asp Gly Tyr Ser Gly Gln Leu Val Cys Ile Ile 
1265                1270                1275                1280 

Asp Asp Ile Gly Gln Asn Thr Thr Asp Glu Asp Trp Ser Asp Phe Cys 
                1285                1290                1295 

Gln Leu Val Ser Gly Cys Pro Met Arg Leu Asn Met Ala Ser Leu Glu 
            1300                1305                1310 

Glu Lys Gly Arg His Phe Ser Ser Pro Phe Ile Ile Ala Thr Ser Asn 
        1315                1320                1325 

Trp Ser Asn Pro Ser Pro Lys Thr Val Tyr Val Lys Glu Ala Ile Asp 
    1330                1335                1340 

Arg Arg Leu His Phe Lys Val Glu Val Lys Pro Ala Ser Phe Phe Lys 
1345                1350                1355                1360 

Asn Pro His Asn Asp Met Leu Asn Val Asn Leu Ala Lys Thr Asn Asp 
                1365                1370                1375 

Ala Ile Lys Asp Met Ser Cys Val Asp Leu Ile Met Asp Gly His Asn 
            1380                1385                1390 

Ile Ser Leu Met Asp Leu Leu Ser Ser Leu Val Met Thr Val Glu Ile 
        1395                1400                1405 

Arg Lys Gln Asn Met Ser Glu Phe Met Glu Leu Trp Ser Gln Gly Ile 
    1410                1415                1420 

Ser Asp Asp Asp Asn Asp Ser Ala Val Ala Glu Phe Phe Gln Ser Phe 
1425                1430                1435                1440 

Pro Ser Gly Glu Pro Ser Asn Trp Lys Leu Ser Ser Phe Phe Gln Ser 
                1445                1450                1455 

Val Thr Asn His Lys Trp Val Ala Val Gly Ala Ala Val Gly Ile Leu 
            1460                1465                1470 

Gly Val Leu Val Gly Gly Trp Phe Val Tyr Lys His Phe Ser Arg Lys 
        1475                1480                1485 

Glu Glu Glu Pro Ile Pro Ala Glu Gly Val Tyr His Gly Val Thr Lys 
    1490                1495                1500 

Pro Lys Gln Val Ile Lys Leu Asp Ala Asp Pro Val Glu Ser Gln Ser 
1505                1510                1515                1520 

Thr Leu Glu Ile Ala Gly Leu Val Arg Lys Asn Leu Val Gln Phe Gly 
                1525                1530                1535 

Val Gly Glu Lys Asn Gly Cys Val Arg Trp Val Met Asn Ala Leu Gly 
            1540                1545                1550 

Val Lys Asp Asp Trp Leu Leu Val Pro Ser His Ala Tyr Lys Phe Glu 
        1555                1560                1565 

Lys Asp Tyr Glu Met Met Glu Phe Tyr Phe Asn Arg Gly Gly Thr Tyr 
    1570                1575                1580 

Tyr Ser Ile Ser Ala Gly Asn Val Val Ile Gln Ser Leu Asp Val Gly 
1585                1590                1595                1600 

Phe Gln Asp Val Val Leu Met Lys Val Pro Thr Ile Pro Lys Phe Arg 
                1605                1610                1615 

Asp Ile Thr Gln His Phe Ile Lys Lys Gly Asp Val Pro Arg Ala Leu 
            1620                1625                1630 

Asn Arg Leu Ala Thr Leu Val Thr Thr Val Asn Gly Thr Pro Met Leu 
        1635                1640                1645 

Ile Ser Glu Gly Pro Leu Lys Met Glu Glu Lys Ala Thr Tyr Val His 
    1650                1655                1660 

Lys Lys Asn Asp Gly Thr Thr Val Asp Leu Thr Val Asp Gln Ala Trp 
1665                1670                1675                1680 

Arg Gly Lys Gly Glu Gly Leu Pro Gly Met Cys Gly Gly Ala Leu Val 
                1685                1690                1695 

Ser Ser Asn Gln Ser Ile Gln Asn Ala Ile Leu Gly Ile His Val Ala 
            1700                1705                1710 

Gly Gly Asn Ser Ile Leu Val Ala Lys Leu Val Thr Gln Glu Met Phe 
        1715                1720                1725 

Gln Asn Ile Asp Lys Lys Ile Glu Ser Gln Arg Ile Met Lys Val Glu 
    1730                1735                1740 

Phe Thr Gln Cys Ser Met Asn Val Val Ser Lys Thr Leu Phe Arg Lys 
1745                1750                1755                1760 

Ser Pro Ile His His His Ile Asp Lys Thr Met Ile Asn Phe Pro Ala 
                1765                1770                1775 

Ala Met Pro Phe Ser Lys Ala Glu Ile Asp Pro Met Ala Met Met Leu 
            1780                1785                1790 

Ser Lys Tyr Ser Leu Pro Ile Val Glu Glu Pro Glu Asp Tyr Lys Glu 
        1795                1800                1805 

Ala Ser Val Phe Tyr Gln Asn Lys Ile Val Gly Lys Thr Gln Leu Val 
    1810                1815                1820 

Asp Asp Phe Leu Asp Leu Asp Met Ala Ile Thr Gly Ala Pro Gly Ile 
1825                1830                1835                1840 

Asp Ala Ile Asn Met Asp Ser Ser Pro Gly Phe Pro Tyr Val Gln Glu 
                1845                1850                1855 

Lys Leu Thr Lys Arg Asp Leu Ile Trp Leu Asp Glu Asn Gly Leu Leu 
            1860                1865                1870 

Leu Gly Val His Pro Arg Leu Ala Gln Arg Ile Leu Phe Asn Thr Val 
        1875                1880                1885 

Met Met Glu Asn Cys Ser Asp Leu Asp Val Val Phe Thr Thr Cys Pro 
    1890                1895                1900 

Lys Asp Glu Leu Arg Pro Leu Glu Lys Val Leu Glu Ser Lys Thr Arg 
1905                1910                1915                1920 

Ala Ile Asp Ala Cys Pro Leu Asp Tyr Thr Ile Leu Cys Arg Met Tyr 
                1925                1930                1935 

Trp Gly Pro Ala Ile Ser Tyr Phe His Leu Asn Pro Gly Phe His Thr 
            1940                1945                1950 

Gly Val Ala Ile Gly Ile Asp Pro Asp Arg Gln Trp Asp Glu Leu Phe 
        1955                1960                1965 

Lys Thr Met Ile Arg Phe Gly Asp Val Gly Leu Asp Leu Asp Phe Ser 
    1970                1975                1980 

Ala Phe Asp Ala Ser Leu Ser Pro Phe Met Ile Arg Glu Ala Gly Arg 
1985                1990                1995                2000 

Ile Met Ser Glu Leu Ser Gly Thr Pro Ser His Phe Gly Thr Ala Leu 
                2005                2010                2015 

Ile Asn Thr Ile Ile Tyr Ser Lys His Leu Leu Tyr Asn Cys Cys Tyr 
            2020                2025                2030 

His Val Cys Gly Ser Met Pro Ser Gly Ser Pro Cys Thr Ala Leu Leu 
        2035                2040                2045 

Asn Ser Ile Ile Asn Asn Ile Asn Leu Tyr Tyr Val Phe Ser Lys Ile 
    2050                2055                2060 

Phe Gly Lys Ser Pro Val Phe Phe Cys Gln Ala Leu Arg Ile Leu Cys 
2065                2070                2075                2080 

Tyr Gly Asp Asp Val Leu Ile Val Phe Ser Arg Asp Val Gln Ile Asp 
                2085                2090                2095 

Asn Leu Asp Leu Ile Gly Gln Lys Ile Val Asp Glu Phe Lys Lys Leu 
            2100                2105                2110 

Gly Met Thr Ala Thr Ser Ala Asp Lys Asn Val Pro Gln Leu Lys Pro 
        2115                2120                2125 

Val Ser Glu Leu Thr Phe Leu Lys Arg Ser Phe Asn Leu Val Glu Asp 
    2130                2135                2140 

Arg Ile Arg Pro Ala Ile Ser Glu Lys Thr Ile Trp Ser Leu Met Ala 
2145                2150                2155                2160 

Trp Gln Arg Ser Asn Ala Glu Phe Glu Gln Asn Leu Glu Asn Ala Gln 
                2165                2170                2175 

Trp Phe Ala Phe Met His Gly Tyr Glu Phe Tyr Gln Lys Phe Tyr Tyr 
            2180                2185                2190 

Phe Val Gln Ser Cys Leu Glu Lys Glu Met Ile Glu Tyr Arg Leu Lys 
        2195                2200                2205 

Ser Tyr Asp Trp Trp Arg Met Arg Phe Tyr Asp Gln Cys Phe Ile Cys 
    2210                2215                2220 

Asp Leu Ser 
2225 

 
           
             13  
             9416  
             DNA  
             Artificial Sequence  
             
               Hepatitis C virus sequence  
             
           
            13 

gccagccccc tgatgggggc gacactccac catgaatcac tcccctgtga ggaactactg     60 

tcttcacgca gaaagcgtct agccatggcg ttagtatgag tgtcgtgcag cctccaggac    120 

cccccctccc gggagagcca tagtggtctg cggaaccggt gagtacaccg gaattgccag    180 

gacgaccggg tcctttcttg gataaacccg ctcaatgcct ggagatttgg gcgtgccccc    240 

gcaagactgc tagccgagta gtgttgggtc gcgaaaggcc ttgtggtact gcctgatagg    300 

gtgcttgcga gtgccccggg aggtctcgta gaccgtgcac catgagcacg aatcctaaac    360 

ctcaaagaaa aaccaaacgt aacaccaacc gtcgcccaca ggacgtcaag ttcccgggtg    420 

gcggtcagat cgttggtgga gtttacttgt tgccgcgcag gggccctaga ttgggtgtgc    480 

gcgcgacgag gaagacttcc gagcggtcgc aacctcgagg tagacgtcag cctatcccca    540 

aggcacgtcg gcccgagggc aggacctggg ctcagcccgg gtacccttgg cccctctatg    600 

gcaatgaggg ttgcgggtgg gcgggatggc tcctgtctcc ccgtggctct cggcctagct    660 

ggggccccac agacccccgg cgtaggtcgc gcaatttggg taaggtcatc gataccctta    720 

cgtgcggctt cgccgacctc atggggtaca taccgctcgt cggcgcccct cttggaggcg    780 

ctgccagggc cctggcgcat ggcgtccggg ttctggaaga cggcgtgaac tatgcaacag    840 

ggaaccttcc tggttgctct ttctctatct tccttctggc cctgctctct tgcctgactg    900 

tgcccgcttc agcctaccaa gtgcgcaatt cctcggggct ttaccatgtc accaatgatt    960 

gccctaactc gagtgttgtg tacgaggcgg ccgatgccat cctgcacact ccggggtgtg   1020 

tcccttgcgt tcgcgagggt aacgcctcga ggtgttgggt ggcggtgacc cccacggtgg   1080 

ccaccaggga cggcaaactc cccacaacgc agcttcgacg tcatatcgat ctgcttgtcg   1140 

ggagcgccac cctctgctcg gccctctacg tgggggacct gtgcgggtct gtctttcttg   1200 

ttggtcaact gtttaccttc tctcccaggc accactggac gacgcaagac tgcaattgtt   1260 

ctatctatcc cggccatata acgggtcatc gcatggcatg gaatatgatg atgaactggt   1320 

cccctacggc agcgttggtg gtagctcagc tgctccgaat cccacaagcc atcatggaca   1380 

tgatcgctgg cgcccactgg ggagtcctgg cgggcataaa gtatttctcc atggtgggga   1440 

actgggcgaa ggtcctggta gtgctgctgc tatttgccgg cgtcgacgcg gaaacccacg   1500 

tcaccggggg aaatgccggc cgcaccacgg ctgggcttgt tggtctcctt acaccaggcg   1560 

ccaagcagaa catccaactg atcaacacca acggcagttg gcacatcaat agcacggcct   1620 

tgaactgcaa tgaaagcctt aacaccggct ggttagcagg gctcttctat cagcacaaat   1680 

tcaactcttc aggctgtcct gagaggttgg ccagctgccg acgccttacc gattttgccc   1740 

agggctgggg tcctatcagt tatgccaacg gaagcggcct cgacgaacgc ccctactgct   1800 

ggcactaccc tccaagacct tgtggcattg tgcccgcaaa gagcgtgtgt ggcccggtat   1860 

attgcttcac tcccagcccc gtggtggtgg gaacgaccga caggtcgggc gcgcctacct   1920 

acagctgggg tgcaaatgat acggatgtct tcgtccttaa caacaccagg ccaccgctgg   1980 

gcaattggtt cggttgtacc tggatgaact caactggatt caccaaagtg tgcggagcgc   2040 

ccccttgtgt catcggaggg gtgggcaaca acaccttgct ctgccccact gattgcttcc   2100 

gcaaatatcc ggaagccaca tactctcggt gcggctccgg tcccaggatt acacccaggt   2160 

gcatggtcga ctacccgtat aggctttggc actatccttg taccatcaat tacaccatat   2220 

tcaaagtcag gatgtacgtg ggaggggtcg agcacaggct ggaagcggcc tgcaactgga   2280 

cgcggggcga acgctgtgat ctggaagaca gggacaggtc cgagctcagc ccgttgctgc   2340 

tgtccaccac acagtggcag gtccttccgt gttctttcac gaccctgcca gccttgtcca   2400 

ccggcctcat ccacctccac cagaacattg tggacgtgca gtacttgtac ggggtagggt   2460 

caagcatcgc gtcctgggcc attaagtggg agtacgtcgt tctcctgttc cttctgcttg   2520 

cagacgcgcg cgtctgttcc tgcttgtgga tgatgttact catatcccaa gcggaggcgg   2580 

ctttggagaa cctcgtaata ctcaatgcag catccctggc cgggacgcat ggtcttgtgt   2640 

ccttcctcgt gttcttctgc tttgcgtggt atctgaaggg taggtgggtg cccggagcgg   2700 

tctacgccct ctacgggatg tggcctctcc tcctgctcct gctggcgttg cctcagcggg   2760 

catacgcact ggacacggag gtggccgcgt cgtgtggcgg cgttgttctt gtcgggttaa   2820 

tggcgctgac tctgtcgcca tattacaagc gctatatcag ctggtgcatg tggtggcttc   2880 

agtattttct gaccagagta gaagcgcaac tgcacgtgtg ggttcccccc ctcaacgtcc   2940 

ggggggggcg cgatgccgtc atcttactca cgtgtgtagt acacccggcc ctggtatttg   3000 

acatcaccaa actactcctg gccatcttcg gacccctttg gattcttcaa gccagtttgc   3060 

ttaaagtccc ctacttcgtg cgcgttcaag gccttctccg gatctgcgcg ctagcgcgga   3120 

agatagccgg aggtcattac gtgcaaatgg ccatcatcaa gttaggggcg cttactggca   3180 

cctgtgtgta taaccatctc gctcctcttc gagactgggc gcacaacggc ctgcgagatc   3240 

tggccgtggc tgtggaacca gtcgtcttct cccgaatgga gaccaagctc atcacgtggg   3300 

gggcagatac cgccgcgtgc ggtgacatca tcaacggctt gcccgtctct gcccgtaggg   3360 

gccaggagat actgcttggg ccagccgacg gaatggtctc caaggggtgg aggttgctgg   3420 

cgcccatcac ggcgtacgcc cagcagacga gaggcctcct agggtgtata atcaccagcc   3480 

tgactggccg ggacaaaaac caagtggagg gtgaggtcca gatcgtgtca actgctaccc   3540 

agaccttcct ggcaacgtgc atcaatgggg tatgctggac tgtctaccac ggggccggaa   3600 

cgaggaccat cgcatcaccc aagggtcctg tcatccagac gtataccaat gtggatcaag   3660 

acctcgtggg ctggcccgct cctcaaggtt cccgctcatt gacaccctgc acctgcggct   3720 

cctcggacct ttacctggtc acgaggcacg ccgatgtcat tcccgtgcgc cggcgaggtg   3780 

atagcagggg tagcctgctt tcgccccggc ccatttccta cttgaaaggc tcctcggggg   3840 

gtccgctgtt gtgccccacg ggacacgccg tgggcctatt cagggccgcg gtgtgcaccc   3900 

gtggagtggc taaggcggtg gactttatcc ctgtggagaa cctagagaca accatgagat   3960 

ccccggtgtt cacggacaac tcctctccac cagcagtgcc ccagagcttc caggtggccc   4020 

acctgcatgc tcccaccggc agcggtaaga gcaccaaggt cccggctgcg tacgcagcca   4080 

agggctacaa ggtgttggtg ctcaacccct ctgttgctgc aacactgggc tttggtgctt   4140 

acatgtccaa ggcccatggg gttgatccta atatcaggac cggggtgaga acaattacca   4200 

ctggcagccc catcacgtac tccacctacg gcaagttcct tgccgacgcc gggtgctcag   4260 

gaggtgctta tgacataata atttgtgacg agtgccactc cacggatgcc acatccatct   4320 

cgggcatcgg cactgtcctt gaccaagcag agactgcggg ggcgagactg gttgtgctcg   4380 

ccactgctac ccctccgggc tccgtcactg tgtcccatcc taacatcgag gaggttgctc   4440 

tgtccaccac cggagagatc cccttttacg gcaaggctat ccccctcgag gtgatcaagg   4500 

ggggaagaca tctcatcttc tgccactcaa agaagaagtg cgacgagctc gccgcgaagc   4560 

tggtcgcatt gggcatcaat gccgtggcct actaccgcgg tcttgacgtg tctgtcatcc   4620 

cgaccagcgg cgatgttgtc gtcgtgtcga ccgatgctct catgactggc tttaccggcg   4680 

acttcgactc tgtgatagac tgcaacacgt gtgtcactca gacagtcgat tttagccttg   4740 

accctacctt taccattgag acaaccacgc tcccccagga tgctgtctcc aggactcaac   4800 

gccggggcag gactggcagg gggaagccag gcatctatag atttgtggca ccgggggagc   4860 

gcccctccgg catgttcgac tcgtccgtcc tctgtgagtg ctatgacgcg ggctgtgctt   4920 

ggtatgagct cacgcccgcc gagactacag ttaggctacg agcgtacatg aacaccccgg   4980 

ggcttcccgt gtgccaggac catcttggat tttgggaggg cgtctttacg ggcctcactc   5040 

atatagatgc ccactttcta tcccagacaa agcagagtgg ggagaacttt ccttacctgg   5100 

tagcgtacca agccaccgtg tgcgctaggg ctcaagcccc tcccccatcg tgggaccaga   5160 

tgcggaagtg tttgatccgc cttaaaccca ccctccatgg gccaacaccc ctgctataca   5220 

gactgggcgc tgttcagaat gaagtcaccc tgacgcaccc aatcaccaaa tacatcatga   5280 

catgcatgtc ggccgacctg gaggtcgtca cgagcacctg ggtgctcgtt ggcggcgtcc   5340 

tggctgctct ggccgcgtat tgcctgtcaa caggctgcgt ggtcatagtg ggcaggatcg   5400 

tcttgtccgg gaagccggca attatacctg acagggaggt tctctaccag gagttcgatg   5460 

agatggaaga gtgctctcag cacttaccgt acatcgagca agggatgatg ctcgctgagc   5520 

agttcaagca gaaggccctc ggcctcctgc agaccgcgtc ccgccatgca gaggttatca   5580 

cccctgctgt ccagaccaac tggcagaaac tcgaggtctt ttgggcgaag cacatgtgga   5640 

atttcatcag tgggatacaa tacttggcgg gcctgtcaac gctgcctggt aaccccgcca   5700 

ttgcttcatt gatggctttt acagctgccg tcaccagccc actaaccact ggccaaaccc   5760 

tcctcttcaa catattgggg gggtgggtgg ctgcccagct cgccgccccc ggtgccgcta   5820 

ccgcctttgt gggcgctggc ttagctggcg ccgcactcga cagcgttgga ctggggaagg   5880 

tcctcgtgga cattcttgca ggctatggcg cgggcgtggc gggagctctt gtggcattca   5940 

agatcatgag cggtgaggtc ccctccacgg aggacctggt caatctgctg cccgccatcc   6000 

tctcacctgg agcccttgca gtcggtgtgg tctttgcatc aatactgcgc cggcgtgttg   6060 

gcccgggcga gggggcagtg caatggatga accggctaat agccttcgcc tcccggggga   6120 

accatgtttc ccccacacac tacgtgccgg agagcgatgc agccgcccgc gtcactgcca   6180 

tactcagcag cctcactgta acccagctcc tgaggcgact gcatcagtgg ataagctcgg   6240 

agtgtaccac tccatgctcc ggttcctggc taagggacat ctgggactgg atatgcgagg   6300 

tgctgagcga ctttaagacc tggctgaaag ccaagctcat gccacaactg cctgggattc   6360 

cctttgtgtc ctgccagcgc gggtataggg gggtctggcg aggagacggc attatgcaca   6420 

ctcgctgcca ctgtggagct gagatcactg gacatgtcaa aaacgggacg atgaggatcg   6480 

tcggtcctag gacctgcaag aacatgtgga gtgggacgtt cttcattaat gcctacacca   6540 

cgggcccctg tactcccctt cctgcgccga actataagtt cgcgctgtgg agggtgtctg   6600 

cagaggaata cgtggagata aggcgggtgg gggacttcca ctacgtatcg ggcatgacta   6660 

ctgacaatct caaatgcccg tgccagatcc catcgcccga atttttcaca gaattggacg   6720 

gggtgcgcct acataggttt gcgccccctt gcaagccctt gctgcgggag gaggtatcat   6780 

tcagagtagg actccacgag tacccggtgg ggtcgcaatt accttgcgag cccgaaccgg   6840 

acgtagccgt gttgacgtcc atgctcactg atccctccca tataacagca gaggcggccg   6900 

ggagaaggtt ggcgagaggg tcaccccctt ctatggccag ctcctcggct agccagctgt   6960 

ccgctccatc tctcaaggca acttgcaccg ccaaccatga ctcccctgac gccgagctca   7020 

tagaggctaa cctcctgtgg aggcaggaga tgggcggcaa catcaccagg gttgagtcag   7080 

agaacaaagt ggtgattctg gactccttcg atccgcttgt ggcagaggag gatgagcggg   7140 

aggtctccgt acccgcagaa attctgcgga agtctcggag attcgcccca gccctgcccg   7200 

tctgggcgcg gccggactac aaccccctgc tagtagagac gtggaaaaag cctgactacg   7260 

aaccacctgt ggtccatggc tgcccgctac cacctccacg gtcccctcct gtgcctccgc   7320 

ctcggaaaaa gcgtacggtg gtcctcaccg aatcaaccct acctactgcc ttggccgagc   7380 

ttgccaccaa aagttttggc agctcctcaa cttccggcat tacgggcgac aatacgacaa   7440 

catcctctga gcccgcccct tctggctgcc cccccgactc cgacgttgag tcctattctt   7500 

ccatgccccc cctggagggg gagcctgggg atccggatct cagcgacggg tcatggtcga   7560 

cggtcagtag tggggccgac acggaagatg tcgtgtgctg ctcaatgtct tattcctgga   7620 

caggcgcact cgtcaccccg tgcgctgcgg aggaacaaaa actgcccatc aacgcactga   7680 

gcaactcgtt gctacgccat cacaatctgg tgtattccac cacttcacgc agtgcttgcc   7740 

aaaggaagaa gaaagtcaca tttgacagac tgcaagttct ggacagccat taccaggacg   7800 

tgctcaagga ggtcaaagca gcggcgtcaa aagtgaaggc taacttgcta tccgtagagg   7860 

aagcttgcag cctggcgccc ccacattcag ccaaatccaa gtttggctat ggggcaaaag   7920 

acgtccgttg ccatgccaga aaggccgtag cccacatcaa ctccgtgtgg aaagaccttc   7980 

tggaagacag tgtaacacca atagacacta ccatcatggc caagaacgag gttttctgcg   8040 

ttcagcctga gaaggggggt cgtaagccag ctcgtctcat cgtgttcccc gacctgggcg   8100 

tgcgcgtgtg cgagaagatg gccctgtacg acgtggttag caagctcccc ttggccgtga   8160 

tgggaagctc ctacggattc caatactcac caggacagcg ggttgaattc ctcgtgcaag   8220 

cgtggaagtc caagaagacc ccgatggggc tctcgtatga tacccgctgt tttgactcca   8280 

cagtcactga gagcgacatc cgtacggagg aggcaattta ccaatgttgt gacctggacc   8340 

cccaagcccg cgtggccatc aagtccctca ctgagaggct ttatgttggg ggccctctta   8400 

ctaattcaag gggggaaaac tgcggctacc gcaggtgccg cgcgagcaga gtactgacaa   8460 

ctagctgtgg taacaccctc actcgctaca tcaaggcccg ggcagcctgt cgagccgcag   8520 

ggctccagga ctgcaccatg ctcgtgtgtg gcgacgactt agtcgttatc tgtgaaagtg   8580 

cgggggtcca ggaggacgcg gcgagcctga gagccttcac ggaggctatg accaggtact   8640 

ccgccccccc cggggacccc ccacaaccag aatacgactt ggagcttata acatcatgct   8700 

cctccaacgt gtcagtcgcc cacgacggcg ctggaaagag ggtctactac cttacccgtg   8760 

accctacaac ccccctcgcg agagccgcgt gggagacagc aagacacact ccagtcaatt   8820 

cctggctagg caacataatc atgtttgccc ccacactgtg ggcgaggatg atactgatga   8880 

cccacttctt tagcgtcctc atagccaggg atcagcttga acaggctctc aactgcgaga   8940 

tctacggagc ctgctactcc atagaaccac tggatctacc tccaatcatt caaagactcc   9000 

atggcctcag cgcattttca ctccacagtt actctccagg tgaaattaat agggtggccg   9060 

catgcctcag aaaacttggg gtcccgccct tgcgagcttg gagacaccgg gcctggagcg   9120 

tccgcgctag gcttctggcc agaggaggca aggctgccat atgtggcaag tacctcttca   9180 

actgggcagt aagaacaaag ctcaaactca ctccgataac ggccgctggc cggctggact   9240 

tgtccggctg gttcacggct ggctacagcg ggggagacat ttatcacagc gtgtctcatg   9300 

cccggccccg ctggttctgg ttttgcctac tcctgcttgc tgcaggggta ggcatctacc   9360 

tcctccccaa ccgatgaaga ttgggctaac cactccaggc caataggcca ttccct       9416 

 
           
             14  
             3182  
             DNA  
             Artificial Sequence  
             
               Hepatitis B virus sequence  
             
           
            14 

aattccacaa ccttccacca aactctgcaa gatcccagag tgagaggcct gtatttccct     60 

gctggtggct ccagttcagg aacagtaaac cctgttctga ctactgcctc tcccttatcg    120 

tcaatcttct cgaggattgg ggaccctgcg ctgaacatgg agaacatcac atcaggattc    180 

ctaggacccc ttctcgtgtt acaggcgggg tttttcttgt tgacaagaat cctcacaata    240 

ccgcagagtc tagactcgtg gtggacttct ctcaattttc tagggggaac taccgtgtgt    300 

cttggccaaa attcgcagtc cccaacctcc aatcactcac caacctcttg tcctccaact    360 

tgtcctggtt atcgctggat gtgtctgcgg cgttttatca tcttcctctt catcctgctg    420 

ctatgcctca tcttcttgtt ggttcttctg gactatcaag gtatgttgcc cgtttgtcct    480 

ctaattccag gatcctcaac aaccagcacg ggaccatgcc ggacctgcat gactactgct    540 

caaggaacct ctatgtatcc ctcctgttgc tgtaccaaac cttcggacgg aaattgcacc    600 

tgtattccca tcccatcatc ctgggctttc ggaaaattcc tatgggagtg ggcctcagcc    660 

cgtttctcct ggctcagttt actagtgcca tttgttcagt ggttcgtagg gctttccccc    720 

actgtttggc tttcagttat atggatgatg tggtattggg ggccaagtct gtacagcatc    780 

ttgagtccct ttttaccgct gttaccaatt ttcttttgtc tttgggtata catttaaacc    840 

ctaacaaaac aaagagatgg ggttactctc taaattttat gggttatgtc attggatgtt    900 

atgggtcctt gccacaagaa cacatcatac aaaaaatcaa agaatgtttt agaaaacttc    960 

ctattaacag gcctattgat tggaaagtat gtcaacgaat tgtgggtctt ttgggttttg   1020 

ctgccccttt tacacaatgt ggttatcctg cgttgatgcc tttgtatgca tgtattcaat   1080 

ctaagcaggc tttcactttc tcgccaactt acaaggcctt tctgtgtaaa caatacctga   1140 

acctttaccc cgttgcccgg caacggccag gtctgtgcca agtgtttgct gacgcaaccc   1200 

ccactggctg gggcttggtc atgggccatc agcgcatgcg tggaaccttt tcggctcctc   1260 

tgccgatcca tactgcggaa ctcctagccg cttgttttgc tcgcagcagg tctggagcaa   1320 

acattatcgg gactgataac tctgttgtcc tatcccgcaa atatacatcg tttccatggc   1380 

tgctaggctg tgctgccaac tggatcctgc gcgggacgtc ctttgtttac gtcccgtcgg   1440 

cgctgaatcc tgcggacgac ccttctcggg gtcgcttggg actctctcgt ccccttctcc   1500 

gtctgccgtt ccgaccgacc acggggcgca cctctcttta cgcggactcc ccgtctgtgc   1560 

cttctcatct gccggaccgt gtgcacttcg cttcacctct gcacgtcgca tggagaccac   1620 

cgtgaacgcc caccaaatat tgcccaaggt cttacataag aggactcttg gactctcagc   1680 

aatgtcaacg accgaccttg aggcatactt caaagactgt ttgtttaaag actgggagga   1740 

gttgggggag gagattaggt taaaggtctt tgtactagga ggctgtaggc ataaattggt   1800 

ctgcgcacca gcaccatgca actttttcac ctctgcctaa tcatctcttg ttcatgtcct   1860 

actgttcaag cctccaagct gtgccttggg tggctttggg gcatggacat cgacccttat   1920 

aaagaatttg gagctactgt ggagttactc tcgtttttgc cttctgactt ctttccttca   1980 

gtacgagatc ttctagatac cgcctcagct ctgtatcggg aagccttaga gtctcctgag   2040 

cattgttcac ctcaccatac tgcactcagg caagcaattc tttgctgggg ggaactaatg   2100 

actctagcta cctgggtggg tgttaatttg gaagatccag cgtctagaga cctagtagtc   2160 

agttatgtca acactaatat gggcctaaag ttcaggcaac tcttgtggtt tcacatttct   2220 

tgtctcactt ttggaagaga aacagttata gagtatttgg tgtctttcgg agtgtggatt   2280 

cgcactcctc cagcttatag accaccaaat gcccctatcc tatcaacact tccggagact   2340 

actgttgtta gacgacgagg caggtcccct agaagaagaa ctccctcgcc tcgcagacga   2400 

aggtctcaat cgccgcgtcg cagaagatct caatctcggg aatctcaatg ttagtattcc   2460 

ttggactcat aaggtgggga actttactgg gctttattct tctactgtac ctgtctttaa   2520 

tcctcattgg aaaacaccat cttttcctaa tatacattta caccaagaca ttatcaaaaa   2580 

atgtgaacag tttgtaggcc cactcacagt taatgagaaa agaagattgc aattgattat   2640 

gcctgccagg ttttatccaa aggttaccaa atatttacca ttggataagg gtattaaacc   2700 

ttattatcca gaacatctag ttaatcatta cttccaaact agacactatt tacacactct   2760 

atggaaggcg ggtatattat ataagagaga aacaacacat agcgcctcat tttgtgggtc   2820 

accatattct tgggaacaag atctacagca tggggcagaa tctttccacc agcaatcctc   2880 

tgggattctt tcccgaccac cagttggatc cagccttcag agcaaacacc gcaaatccag   2940 

attgggactt caatcccaac aaggacacct ggccagacgc caacaaggta ggagctggag   3000 

cattcgggct gggtttcacc ccaccgcacg gaggcctttt ggggtggagc cctcaggctc   3060 

agggcatact acaaactttg ccagcaaatc cgcctcctgc ctccaccaat cgccagtcag   3120 

gaaggcagcc taccccgctg tctccacctt tgagaaacac tcatcctcag gccatgcagt   3180 

gg                                                                  3182 

 
           
             15  
             7478  
             DNA  
             Artificial Sequence  
             
               Hepatitis A virus sequence  
             
           
            15 

ttcaagaggg gtctccggag gtttccggag cccctcttgg aagtccatgg tgaggggact     60 

tgatacctca ccgccgtttg cctaggctat aggctaaatt tccctttccc tgtccctccc    120 

ttatttccct ttgttttgct tgtaaatatt aattcctgca ggttcagggt tctttaatct    180 

gtttctctat aagaacactc aattttcacg ctttctgtct tctttcttcc agggctctcc    240 

ccttgcccta ggctctggcc gttgcgcccg gcggggtcaa ctccatgatt agcatggagc    300 

tgtaggagtc taaattgggg acgcagatgt ttgggacgtc accttgcagt gttaacttgg    360 

ctctcatgaa cctctttgat cttccacaag gggtaggcta cgggtgaaac ctcttaggct    420 

aatacttcta tgaagagatg ctttggatag ggtaacagcg gcggatattg gtgagttgtt    480 

aagacaaaaa ccattcaacg ccggaggact ggctctcatc cagtggatgc attgagtgga    540 

ttgattgtca gggctgtctc taggtttaat ctcagacctc tctgtgctta gggcaaacac    600 

catttggcct taaatgggat cctgtgagag ggggtccctc cattgacagc tggactgttc    660 

tttggggcct tatgtggtgt ttgcctctga ggtactcagg ggcatttagg tttttcctca    720 

ttcttaaaca ataatgaata tgtccaaaca aggaattttc cagactgttg ggagtggcct    780 

tgaccacatc ctgtctttgg cagatattga ggaagagcaa atgattcagt ccgttgatag    840 

gactgcagtg actggagctt cttacttcac ttctgtggac caatcttcag ttcatactgc    900 

tgaggttggc tcacatcaaa ttgaaccttt gaaaacctct gttgataaac ctggttctaa    960 

gaaaactcag ggggaaaagt ttttcctgat tcattctgct gattggctca ctacacatgc   1020 

tctctttcat gaagttgcaa aattggatgt ggtgaaacta ctgtataatg agcagtttgc   1080 

cgtccaaggt ttgttgagat accatacata tgcaagattt ggcattgaga ttcaagttca   1140 

gataaatccc acaccctttc agcaaggagg actaatttgt gccatggttc ctggtgacca   1200 

aagttatggt tcaatagcat ccttgactgt ttatcctcat ggtctgttaa attgcaatat   1260 

caacaatgta gttagaataa aggttccatt tatttatact agaggtgctt atcattttaa   1320 

agatccacag tacccagttt gggaattgac aatcagagtt tggtcagagt tgaatattgg   1380 

aacaggaact tcagcttaca cttcactcaa tgttttagct aggtttacag atttggagtt   1440 

gcatggatta actcctcttt ctacacagat gatgagaaat gaatttaggg tcagtactac   1500 

tgaaaatgtt gtaaatttgt caaattatga agatgcaagg gcaaaaatgt cttttgcttt   1560 

ggatcaggaa gattggaagt ctgatccttc ccaaggtggt ggaattaaaa ttactcattt   1620 

tactacctgg acatccattc caaccttagc tgctcagttt ccatttaatg cttcagattc   1680 

agttggacaa caaattaaag ttattccagt ggacccatac tttttccaaa tgacaaacac   1740 

taatcctgat caaaaatgta taactgcctt ggcctctatt tgtcagatgt tctgcttttg   1800 

gaggggagat cttgtttttg attttcaggt ttttccaacc aaatatcatt caggtagact   1860 

gttgttttgt tttgttcctg ggaatgagtt aatagatgtt actggaatta cattaaaaca   1920 

ggcaactact gctccttgtg cagtgatgga cattacagga gtgcagtcaa ccttgagatt   1980 

tcgtgttcct tggatttctg atacacctta tcgagtgaat aggtacacga agtcagcaca   2040 

tcaaaaaggt gagtacactg ccattgggaa gcttattgtg tattgttata acagactgac   2100 

ttctccttct aatgttgcct ctcatgttag agttaatgtt tatctttcag caattaattt   2160 

ggaatgtttt gctcctcttt accatgctat ggatgttact acacaggttg gagatgattc   2220 

aggaggtttc tcaacaacag tttctacaga gcagaatgtt cctgatcccc aagttgggat   2280 

aacaaccatg agggatttaa aaggaaaagc caatagggga aagatggatg tttcaggagt   2340 

gcaagcacct cgtgggagct atcagcaaca attgaacgat ccagttttag caaagaaagt   2400 

acctgagaca tttcctgaat tgaagcctgg agagtccaga catacatcag atcacatgtc   2460 

tatttataaa ttcatgggaa ggtctcattt tttgtgcact tttactttca attcaaataa   2520 

taaagagtac acatttccaa taaccctgtc ttcgacttct aatcctcctc atggtttacc   2580 

atcaacatta aggtggttct tcaatttgtt tcagttgtat agaggaccat tggatttaac   2640 

aattataatc acaggagcca ctgatgtgga tggtatggcc tggtttactc cagtgggcct   2700 

tgctgtcgac ccttgggtgg aaaaggagtc agctttgtct attgattata aaactgccct   2760 

tggagctgtt agatttaata caagaagaac aggaaacatt caaattagat tgccgtggta   2820 

ttcttatttg tatgccgtgt ctggagcact ggatggcttg ggggataaga cagattctac   2880 

atttggattg tttctattcg agattgcaaa ttacaatcat tctgatgaat atttgtcctt   2940 

cagttgttat ttgtctgtca cagagcaatc agagttctat tttcctagag ctccattaaa   3000 

ttcaaatgct atgttgtcca ctgaatccat gatgagtaga attgcagctg gagacttgga   3060 

gtcatcagtg gatgatccca gatcagagga ggatagaaga tttgagagtc atatagaatg   3120 

taggaaacca tacaaagaat tgagactgga ggttgggaaa caaagactca aatatgctca   3180 

ggaagagtta tcaaatgaag tgcttccacc tcctaggaaa atgaaggggt tattttcaca   3240 

agctaaaatt tctctttttt atactgagga gcatgaaata atgaagtttt cttggagagg   3300 

agtgactgct gatactaggg ctttgagaag atttggattc tctctggctg ctggtagaag   3360 

tgtgtggact cttgaaatgg atgctggagt tcttactgga agattgatca gattgaatga   3420 

tgagaaatgg acagaaatga aggatgataa gattgtttca ttaattgaaa agttcacaag   3480 

caataaatat tggtctaaag tgaattttcc acatggaatg ttggatcttg aagaaattgc   3540 

tgccaattct aaggattttc caaatatgtc tgagacagat ttgtgtttcc tgttacattg   3600 

gctaaatcca aagaaaatca atttagcaga tagaatgctt ggattgtctg gagtgcagga   3660 

aattaaggaa cagggtgttg gactgatagc agagtgtaga actttcttgg attctattgc   3720 

tgggactttg aaatctatga tgtttgggtt tcatcattct gtgactgttg aaattataaa   3780 

tactgtgctt tgttttgtta agagtggaat cctgctttat gtcatacaac aattgaacca   3840 

agatgaacac tctcacataa ttggtttgtt gagagttatg aattatgcag atattggctg   3900 

ttcagttatt tcatgtggta aagttttttc caaaatgtta gaaacagttt ttaattggca   3960 

aatggattct agaatgatgg agctgaggac tcagagcttc tctaattggt taagagatat   4020 

ttgttcagga attactattt ttaaaagttt taaggatgcc atatattggt tatatacaaa   4080 

attgaaggat ttttatgaag taaattatgg caagaaaaag gatattctta atattctcaa   4140 

agataatcag caaaaaatag aaaaagccat tgaagaagca gacaattttt gcattttgca   4200 

aattcaagat gtagagaaat ttgatcagta tcagaaaggg gttgatttaa tacaaaagct   4260 

gagaactgtc cattcaatgg cgcaagttga ccccaatttg ggggttcatt tgtcacctct   4320 

cagagattgc atagcaagag tccaccaaaa gctcaagaat cttggatcta taaatcaggc   4380 

catggtaaca agatgtgagc cagttgtttg ctatttgtat ggcaaaagag ggggagggaa   4440 

aagcttgact tcaattgcat tggcaaccaa aatttgtaaa cactatggtg ttgaacctga   4500 

gaaaaatatt tacaccaaac ctgtggcctc agattattgg gatggatata gtggacaatt   4560 

agtttgcatt attgatgata ttggccaaaa cacaacagat gaagattggt cagatttttg   4620 

tcaattagtg tcaggatgcc caatgagatt gaatatggct tctctagagg agaagggcag   4680 

acatttttcc tctcctttta taatagcaac ttcaaattgg tcaaatccaa gtccaaaaac   4740 

agtttatgtt aaggaagcaa ttgatcgtag gcttcatttt aaggttgaag ttaaacctgc   4800 

ttcatttttt aaaaatcctc acaatgatat gttgaatgtt aatttggcca aaacaaatga   4860 

tgcaattaag gacatgtctt gtgttgattt aataatggat ggacacaata tttcattgat   4920 

ggatttactt agttccttag tgatgacagt tgaaattagg aaacagaata tgagtgaatt   4980 

catggagttg tggtctcagg gaatttcaga tgatgacaat gatagtgcag tggctgagtt   5040 

tttccagtct tttccatctg gtgaaccatc aaattggaag ttatctagtt ttttccaatc   5100 

tgtcactaat cacaagtggg ttgctgtggg agctgcagtt ggcattcttg gagtgcttgt   5160 

gggaggatgg tttgtgtata agcatttttc ccgcaaagag gaagaaccaa ttccagctga   5220 

aggggtttat catggcgtga ctaagcccaa acaagtgatt aaattggatg cagatccagt   5280 

agagtcccag tcaactctag aaatagcagg attagttagg aaaaatctgg ttcagtttgg   5340 

agttggtgag aaaaatggat gtgtgagatg ggtcatgaat gccttaggag tgaaggatga   5400 

ttggttgtta gtaccttctc atgcttataa atttgaaaag gattatgaaa tgatggagtt   5460 

ttacttcaat agaggtggaa cttactattc aatttcagct ggtaatgttg ttattcaatc   5520 

tttagatgtg ggatttcaag atgttgtttt aatgaaggtt cctacaattc ccaagtttag   5580 

agatattact caacacttta ttaagaaagg agatgtgcct agagccttaa atcgcttggc   5640 

aacattagtg acaaccgtta atggaactcc tatgttaatt tctgagggac cattaaagat   5700 

ggaagaaaaa gccacttatg ttcataagaa gaatgatggt actacagttg atttgactgt   5760 

agatcaggca tggagaggaa aaggtgaagg tcttcctgga atgtgtggtg gggccctagt   5820 

gtcatcaaat cagtccatac agaatgcaat tttgggtatt catgttgctg gaggaaattc   5880 

aattcttgtg gcaaagctgg ttactcaaga aatgtttcaa aacattgata agaaaattga   5940 

aagtcagaga ataatgaaag tggaatttac tcaatgttca atgaatgtag tctccaaaac   6000 

gctttttaga aagagtccca ttcatcacca cattgataaa accatgatta attttcctgc   6060 

agctatgcct ttctctaaag ctgaaattga tccaatggct atgatgttgt ccaaatattc   6120 

attacctatt gtggaggaac cagaggatta caaggaagct tcagtttttt atcaaaacaa   6180 

aatagtaggc aagactcagc tagttgatga ctttttagat cttgatatgg ctattacagg   6240 

ggctccaggc attgatgcta tcaatatgga ttcatctcct gggtttcctt atgttcaaga   6300 

aaaattgacc aaaagagatt taatttggtt ggatgaaaat ggtttgctgt taggagttca   6360 

cccaagattg gcccagagaa ttttatttaa tactgtcatg atggaaaatt gttctgactt   6420 

agatgttgtt tttacaactt gtccaaaaga tgaattgaga ccattagaga aagttttgga   6480 

atcaaaaaca agagccattg atgcttgtcc tttggattat acaattctat gtcgaatgta   6540 

ttggggtcca gctatcagtt atttccattt gaatccaggg tttcacacag gtgttgctat   6600 

tggcatagat cctgatagac agtgggatga attatttaaa acaatgataa gatttggaga   6660 

tgttggtctt gatttagatt tctctgcttt tgatgccagt cttagtccat ttatgattag   6720 

ggaagcaggt agaatcatga gtgaattatc tggaacacca tctcattttg gaacagctct   6780 

tatcaatact atcatttatt ctaaacatct gctgtacaac tgttgttatc atgtttgtgg   6840 

ttcaatgcct tctgggtctc cttgcacagc tttgttgaat tcaattatta ataatattaa   6900 

tctgtattat gtgttttcta aaatatttgg aaagtctcca gttttctttt gtcaagcttt   6960 

gaggatcctt tgttacggag atgatgtttt gatagttttt tccagagatg ttcaaattga   7020 

caatcttgac ttgattggac agaaaattgt agatgagttc aaaaaacttg gcatgacagc   7080 

cacctcagct gataaaaatg tgcctcaact gaagccagtt tcagaattga cttttctcaa   7140 

aagatctttc aatttggtgg aggatagaat tagacctgca atttcagaaa agacaatttg   7200 

gtctttgatg gcttggcaga gaagtaacgc tgagtttgag cagaatttag aaaatgctca   7260 

gtggtttgct tttatgcatg gctatgagtt ctatcagaaa ttttattatt ttgttcagtc   7320 

ctgtttggag aaagagatga tagaatatag acttaaatct tatgattggt ggagaatgag   7380 

attttatgac cagtgtttca tttgtgacct ttcatgattt gtttaaacaa attttcttac   7440 

tctttctgag gtttgtttat ttcttttgtc cgctaact                           7478 

5 

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