MALARIA VACCINE

Abstract

The present invention provides a particle comprising a polypeptide and at least one malaria antigen, and a composition or vaccine comprising thereof, its use in medicine, particularly in the prevention or treatment of malaria infections.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This is a continuation of U.S. application Ser. No. 14/294,968 filed Jun. 3, 2014, which claims benefit from U.S. Provisional Application No. 61/830,436, filed Jun. 3, 2013, and U.S. Provisional Application No. 61/906,583, filed Nov. 20, 2013. The entire disclosures of the prior applications are considered part of the disclosure of the accompanying continuation, and are hereby incorporated by reference.

TECHNICAL FIELD

[0002] The present invention relates to a particle comprising a polypeptide and at least one malaria antigen, and a composition or vaccine comprising thereof, its use in medicine, particularly in the prevention or treatment of malaria.

BACKGROUND

[0003] Malaria is one of the world's most prevalent serious infectious diseases, with approximately 250 million cases and 1 million deaths per year (WHO, 2009). Mortality is primarily in children under the age of five and in pregnant women. Every 45 seconds, an African child dies of malaria. The disease is transmitted from person to person by infected mosquitoes, so past eradication efforts involved massive insecticide campaigns. These were successful in the Southeast U.S. for example, but failed in most poorly developed tropical countries. Current efforts involve distribution of bednets, particularly bednets impregnated with insecticide, to prevent mosquito bites at night. However, resistance to insecticides and to anti-malarial drugs for both prevention and treatment is rapidly rising. Thus, the need for a malaria vaccine is imperative for protection of millions of people from disease (http://www.globalvaccines.org/content/malaria+vaccine+prog ram/19614).

[0004] Malaria caused by Plasmodium falciparum remains a major public health threat, especially among children and pregnant women in Africa. An effective malaria vaccine would be a valuable tool to reduce the disease burden and could contribute to elimination of malaria in some regions of the world. Current malaria vaccine candidates are directed against human and mosquito stages of the parasite life cycle, but thus far, relatively few proteins have been studied for potential vaccine development.

[0005] The most advanced vaccine candidate, RTS,S, conferred partial protection against malaria in phase II clinical trials and is currently being evaluated in a phase III trial in Africa. (The Journal of Clinical Investigation 120(12) 4168-4178, 2010).

[0006] The CSP is the predominant surface antigen on sporozoites. CSP is composed of an N-terminal region that binds heparin sulfate proteoglycans (RI), a central region containing a four-amino-acid (NPNA) repeat, and a GPI-anchored C-terminal region containing a thrombospondin-like domain (RII). The region of the CSP included in the RTS,S vaccine includes the last 16 NPNA repeats and the entire flanking C-terminus. HBsAg particles serve as the matrix carrier for RTS,S, 25% of which is fused to the CSP segment (The Journal of Clinical Investigation 120(12) 4168-4178, 2010).

[0007] In a series of phase II clinical trials for RTS,S, 30%-50% of malaria-naive adults immunized with RTS,S were protected against challenge by mosquitoes infected with the homologous P. falciparum clone. In phase II field trials in the Gambia and Kenya, RTS,S conferred short-lived protection against malaria infection in approximately 35% of adults, although results from the Kenya trial did not reach statistical significance. Approximately 30%-50% of children and infants immunized with RTS,S in phase II trials conducted in Mozambique, Tanzania, and Kenya were protected from clinical malaria, however, protection was generally short-lived (The Journal of Clinical Investigation 120(12) 4168-4178, 2010). Results from a pivotal, large-scale Phase III trial, published Nov. 9, 2012, online in the New England Journal of Medicine (NEJM), show that the RTS,S malaria vaccine candidate can help protect African infants against malaria. When compared to immunization with a control vaccine, infants (aged 6-12 weeks at first vaccination) vaccinated with RTS,S had one-third fewer episodes of both clinical and severe malaria and had similar reactions to the injection.

[0008] There are currently no licensed vaccines against malaria. Highly effective malaria vaccine is strongly desired.

[0009] Virus-like particles (VLPs) are multiprotein structures that mimic the organization and conformation of authentic native viruses but lack the viral genome, potentially yielding safer and cheaper vaccine candidates. A handful of prophylactic VLP-based vaccines is currently commercialized worldwide: GlaxoSmithKline's Engerix.RTM. (hepatitis B virus) and Cervarix.RTM. (human papillomavirus), and Merck and Co., Inc.'s Recombivax HB.RTM. (hepatitis B virus) and Gardasil.RTM. (human papillomavirus) are some examples. Other VLP-based vaccine candidates are in clinical trials or undergoing preclinical evaluation, such as, influenza virus, parvovirus, Norwalk and various chimeric VLPs. Many others are still restricted to small-scale fundamental research, despite their success in preclinical tests. The implications of large-scale VLP production are discussed in the context of process control, monitorization and optimization. The main up- and down-stream technical challenges are identified and discussed accordingly. Successful VLP-based vaccine blockbusters are briefly presented concomitantly with the latest results from clinical trials and the recent developments in chimeric VLP-based technology for either therapeutic or prophylactic vaccination (Expert Rev. Vaccines 9(10), 1149-1176, 2010).

[0010] Chikungunya virus (CHIKV) has infected millions of people in Africa, Europe and Asia since this alphavirus reemerged from Kenya in 2004. The severity of the disease and the spread of this epidemic virus present a serious public health threat in the absence of vaccines or antiviral therapies. It is reported that a VLP vaccine for epidemic Chikungunya virus protects non-human primates against infection (Nat Med. 2010 March; 16(3): 334-338). US patent publication No. 2012/0003266 discloses a virus-like particle (VLP) comprising one or more Chikungunya virus structural polypeptides which is useful for formulating a vaccine or antigenic composition for Chikungunya that induces immunity to an infection or at least one symptom thereof. WO2012/106356 discloses modified alphavirus or flavivirus virus-like particles (VLPs) and methods for enhancing production of modified VLPs for use in the prevention or treatment of alphavirus and flavivirus-mediated diseases. (these cited references are herein incorporated by reference).

SUMMARY OF THE INVENTION

[0011] In the first aspect, the present invention provides a particle which is capable of being self-assembled, comprising a polypeptide and at least one malaria antigen, wherein said polypeptide comprises at least one first attachment site and said at least one malaria antigen comprises at least one second attachment site, and wherein said polypeptide and said malaria antigen are linked through said at least one first and said at least one second attachment site.

[0012] In the second aspect, the present invention provides a nucleic acid molecule which is designed for expression of a particle provided in the first aspect of the present invention.

[0013] In the third aspect, the present invention provides a composition or vaccine comprising the particle provided in the first aspect of the present invention and/or the nucleic acid molecule provided in the second aspect of the present invention.

[0014] In the fourth aspect, the present invention provides a method of producing an antibody, comprising contacting the particle provided in the first aspect of the present invention and/or the nucleic acid molecule provided in the second aspect of the present invention to a mammal.

[0015] In the fifth aspect, the present invention provides a method of immunomodulation, a method of treating malaria, a method of inducing and/or enhancing immune response against a malaria antigen in a mammal, comprising administering the composition provided in the third aspect of the present invention to a mammal.

[0016] In sixth aspect, the present invention provides a method of passive immunization against a malaria-causing pathogen, comprising administering the antibody provided in the fourth aspect of the present invention to a mammal.

[0017] In seventh aspect, the present invention provides a method of presenting an antigen on macrophage, comprising contacting the particle provided in the first aspect of the present invention and/or the nucleic acid molecule provided in the second aspect of the present invention to a mammal.

[0018] In eighth aspect, the present invention provides a method for producing the particle provided in the first aspect of the present invention, comprising preparing a vector which is designed for expression of said particle; culturing a cell which is transfected with said vector to express said particle; and recovering said particle.

BRIEF DESCRIPTION OF THE INVENTION

[0019] FIG. 1 shows pVLP74_15 (VLP_CHI 532 NPNA.times.6) vector.

[0020] FIG. 2 shows pVLP78_15 (VLP_CHI 532 NPNA.times.25) vector.

[0021] FIG. 3 shows pVLP74_25 (VLP_VEEV 519 NPNA.times.6) vector.

[0022] FIG. 4 shows that the serum from individual monkeys immunized with Malaria VLPs after 2 weeks induced high titer of antibodies against CSP.

[0023] FIG. 5 shows mean value and SD of the data shown in FIG. 4.

[0024] FIG. 6 shows effects of combined immunization of CHIKV VLP and VEEV VLP on induction of antibodies against CSP. In the figure, Adj indicates adjuvant.

[0025] FIG. 7 shows effects of administered VLP fused with no malaria antigen on induction of antibodies against CSP. In the figure, 4w, 6w, 10w and 14w indicate 4 weeks after immunization, 6 weeks after immunization, 10 weeks after immunization and 14 weeks after immunization, respectively.

[0026] FIG. 8 shows effects of administered VLP fused with malaria antigen on induction of antibodies against CSP. In the figure, 4w, 6w, 10w and 14w indicate 4 weeks after immunization, 6 weeks after immunization, 10 weeks after immunization and 14 weeks after immunization, respectively.

[0027] FIG. 9 shows effects of administered VLP fused with malaria antigen together with adjuvant on induction of antibodies against CSP. In the figure, 4w, 6w, 10w and 14w indicate 4 weeks after immunization, 6 weeks after immunization, 10 weeks after immunization and 14 weeks after immunization, respectively.

[0028] FIG. 10 shows schedule of the experiment.

[0029] FIG. 11 shows detection of 18S malaria DNA by means of PCR.

DETAILED DESCRIPTION OF THE INVENTION

(1) A Particle Comprising a Polypeptide and at Least One Malaria Antigen

[0030] In the first aspect, the present invention provides a particle which is capable of being self-assembled, comprising a polypeptide and at least one malaria antigen, wherein said polypeptide comprises at least one first attachment site and said at least one antigen comprises at least one second attachment site, and wherein said polypeptide and said malaria antigen are linked through said at least one first and said at least one second attachment site.

[0031] As used herein, "a particle which is capable of being self-assembled" refers to a particle formed by at least one constituent which is spontaneously assembled. The constituent may be a polypeptide or non-peptide chemical compound. In one embodiment, "a particle which is capable of being self-assembled" may be a particle comprising or consisting of at least one polypeptide. The at least one polypeptide consists of one or more kinds of peptide. In one embodiment, said particle has a diameter of at least 10 nm, for example, at least 20 nm, preferably at least 50 nm. In one embodiment, molecular weight of said particle is from 100 kDa to 100,000 kDa, preferably from 400 kDa to 30,000 kDa.

[0032] A polypeptide used for the present invention may be spontaneously assembled. The polypeptide may be a virus structural polypeptide. Thus, the particle provided by the present invention may be a virus like particle.

[0033] A virus structural polypeptide may be a naturally occurring viral polypeptide or modified polypeptide thereof. In one embodiment, the modified polypeptide has at least 70%, 75%, 80%, 85%, 90%, 95% or 98% amino acid sequence identity to a naturally occurring viral structural polypeptide including capsid and envelope protein. In one embodiment, the modified polypeptide is a mutant where at most 10% of the amino acids are deleted, substituted, and/or added to a naturally occurring viral structural polypeptide including capsid and envelope protein.

[0034] In one embodiment, virus structural polypeptide used for the present invention consists of or comprises capsid and/or envelope protein or fragment thereof. For example, virus structural polypeptide used for the present invention consists of or comprises capsid and E2 and E1. An antigen may be inserted into E2. In one embodiment, a particle provided by the first aspect of the present invention can be formed by assembling 240 capsids, 240 E1 proteins and 240 E2 proteins where a malaria antigen is inserted into each of E2 proteins.

[0035] Virus structural polypeptide used for the present invention may be derived from Alphavirus or Flavivirus. Thus, the particle provided by the present invention may be a virus like particle derived from Alphavirus or Flavivirus. Examples of Alphavirus and Flavivirus include, but not limited to, Aura virus, Babanki virus, Barmah Forest virus (BFV), Bebaru virus, Cabassou virus, Chikungunya virus (CHIKV), Eastern equine encephalitis virus (EEEV), Eilat virus, Everglades virus, Fort Morgan virus, Getah virus, Highlands J virus, Kyzylagach virus, Mayaro virus, Me Tri virus, Middelburg virus, Mosso das Pedras virus, Mucambo virus, Ndumu virus, O'nyong-nyong virus, Pixuna virus, Rio Negro virus, Ross River virus (RRV), Salmon pancreas disease virus, Semliki Forest virus, Sindbis virus, Southern elephant seal virus, Tonate virus, Trocara virus, Una virus, Venezuelan equine encephalitis virus (VEEV), Western equine encephalitis virus (WEEV), Whataroa virus, West Nile virus, dengue virus, tick-borne encephalitis virus and yellow fever virus.

[0036] Malaria is a disease which human or other animal (e.g. mouse) suffers from. Example of malaria include, but are not limited to, a disease caused by Plasmodium (P.) species including P. falciparum, P. malariae, P.ovale, P.vivax, P.knowlesi, P.berghei, P.chabaudi and P.yoelii

[0037] As used herein, the term "malaria antigen" refers to any antigen or fragment thereof. The term antigen or fragment thereof, means any peptide-based sequence that can be recognized by the immune system and/or that stimulates a cell-mediated immune response and/or stimulates the generation of antibodies.

[0038] According to Scand. J. Immunol. 56, 327-343, 2002, considering the whole parasite life cycle, there are essentially six targets for a malaria vaccine: (1) sporozoites; (2) liver stages; (3) merozoites; (4) infected RBC; (5) parasite toxins; (6) sexual stages.

[0039] Table summarizes the main candidate antigens of each stage identified.

TABLE-US-00001 TABLE 1 Main vaccine candidates from the different phases of Plasmodium life cycle Targets Candidate antigens Sporozoite Circumsporozoite protein (CSP) Thrombospondin-related adhesive protein (TRAP) Sporozoite and liver-stage antigen (SALSA) Sporozoite threonine- and asparagine-rich protein (STARP) Liver stage CSP Liver-stage antigen (LSA)-1 and -3 SALSA STARP Merozoite Merozoite surface protein (MSP)-1, -2, -3, -4 and -5 Erythrocyte-binding antigen (EBA)-175 Apical membrane antigen (AMA)-1 Rhoptry-associated protein (RAP)-1 and -2 Acidic-basic repeat antigen (ABRA) Duffy-binding protein (DBP) (Plasmodium vivax) Blood stage Ring erythrocyte surface antigen (RESA) Serine-rich protein (SERP) Erythrocyte membrane protein (EMP)-1, -2 and -3 Glutamate-rich protein (GLURP) Toxins Glycosilphosphatidylinositol (GPI) Sexual Ps25, Ps28, Ps48/45 and Ps230 stages (Scand. J. Immunol. 56, 327-343, 2002)

[0040] According to the present invention, one or more antigens listed above can be used as long as it is formed to a particle. For example, a circumsporozoite protein and a fragment thereof can be used as an antigen. Examples of circumsporozoite protein include, but are not limited to, Plasmodium falciparum circumsporozoite protein consisting of amino acid sequence described below (SEQ ID No. 56):

TABLE-US-00002 Mmrklailsyssflfvealfqeyqcygsssntrvinelnydnagtn lynelemnyygkqenwyslkknsrslgenddgnnnngdngregkde dkrdgnnedneklrkpkhkklkqpgdgnpdpnanpnvdpnanpnvd pnanpnvdpnanpnanpnanpnanpnanpnanpnanpnanpnanpn anpnanpnanpnanpnanpnanpnvdpnanpnanpnanpnanpnan pnanpnanpnanpnanpnanpnanpnanpnanpnanpnanpnanpn anpnanpnanpnknnqgngqghnmpndpnrnvdenanannavknnn neepsdkhiegylkkiknsistewspcsvtcgngiqvrikpgsank pkdeldyendiekkickmekcssvfnvvnssiglimvlsflflntr.

[0041] In one embodiment, malaria antigen is a Plasmodium falciparum circumsporozoite protein B cell epitope. Example of Plasmodium falciparum circumsporozoite protein B cell epitope may be a repeat sequence of NPNA, including (NPNA).sub.4-30 (i.e. 4.times.NPNA, 5.times.NPNA, 6.times.NPNA, 7.times.NPNA, 8.times.NPNA, 9.times.NPNA, 10.times.NPNA, 11.times.NPNA, 12.times.NPNA, 13.times.NPNA 14.times.NPNA, 15.times.NPNA, 16.times.NPNA, 17.times.NPNA, 18.times.NPNA, 19.times.NPNA, 20.times.NPNA, 21.times.NPNA, 22.times.NPNA, 23.times.NPNA, 24.times.NPNA, 25.times.NPNA, 26.times.NPNA, 27.times.NPNA, 28.times.NPNA, 29.times.NPNA or 30.times.NPNA).

[0042] In one embodiment, malaria antigen is a Plasmodium yoelii circumsporozoite protein B cell epitope including (QGPGAP).sub.3-12.

[0043] In one embodiment, malaria antigen is a Plasmodium vivax circumsporozoite protein B cell epitope including (ANGAGNQPG).sub.1-12.

[0044] In one embodiment, malaria antigen is a Plasmodium malariae circumsporozoite protein B cell epitope including (NAAG).sub.4-30.

[0045] In one embodiment, malaria antigen is a Plasmodium falciparum circumsporozoite protein T cell epitope. Example of Plasmodium falciparum circumsporozoite protein T cell epitope may be EYLNKIQNSLSTEWSPCSVT (SEQ ID No. 44) (EYLNKIQNSLSTEWSPCSVT).sub.1-6 may be also used as a malaria antigen.

[0046] In one embodiment, malaria antigen is a Plasmodium yoelii circumsporozoite protein T cell epitope which is YNRNIVNRLLGDALNGPEEK (SEQ ID No.45). (YNRNIVNRLLGDALNGPEEK).sub.1-6 may be also used as a malaria antigen.

[0047] The present invention addresses one or more of the above needs by providing antigens, vectors encoding the antigens, and antibodies (and antibody-like molecules including aptamers and peptides) that specifically bind to the antigen, together with the uses thereof (either alone or in combination) in the prevention or treatment of malaria infections. As used herein, the term "antibody" refers to molecules which are capable of binding an epitope or antigenic determinant. The term is meant to include whole antibodies and antigen-binding fragments thereof, including single-chain antibodies. Such antibodies include human antigen binding antibody fragments and include, but are not limited to, Fab, Fab' and F(ab')2, Fd, single-chain Fvs (scFv), single-chain antibodies, disulfide-linked Fvs (sdFv) and fragments comprising either a VL or VH domain. The antibodies can be from any animal origin including birds and mammals. Preferably, the antibodies are mammalian e.g. human, murine, rabbit, goat, guinea pig, camel, horse and the like, or other suitable animals e.g. chicken. As used herein, "human" antibodies include antibodies having the amino acid sequence of a human immunoglobulin and include antibodies isolated from human immunoglobulin libraries or from animals transgenic for one or more human immunoglobulins and that do not express endogenous immunoglobulins, as described, for example, in U.S. Pat. No. 5,939,598, the disclosure of which is incorporated herein by reference in its entirety.

[0048] The antigen used for the present invention can be modified polypeptide derived from a naturally occurring protein. The modified polypeptide may be a fragment of the naturally occurring protein. In one embodiment, the modified polypeptide has at least 70%, 75%, 80%, 85%, 90%, 95% or 98% amino acid sequence identity to a polypeptide derived from a naturally occurring protein. In one embodiment, the modified polypeptide derived is a mutant where at most 10% of the amino acids are deleted, substituted, and/or added based on a polypeptide derived from naturally occurring protein.

[0049] In the particle as provided by the present invention, a polypeptide and an antigen may be linked through at least one first attachment site which is present in the polypeptide and at least one second attachment site which is present in the antigen.

[0050] As used herein, each of "a first attachment site" and "a second attachment site" refers to a site where more than one substance is linked each other.

[0051] In one embodiment, the polypeptide and the antigen are directly fused. Alternatively, one or two linkers may intervene between N-terminal residue of the antigen and the polypeptide and/or between C-terminal residue of the antigen and the polypeptide.

[0052] The antigen or the polypeptide can be truncated and replaced by short linkers. In some embodiments, the antigen or the polypeptide include one or more peptide linkers. Typically, a linker consists of from 2 to 25 amino acids. Usually, it is from 2 to 15 amino acids in length, although in certain circumstances, it can be only one, such as a single glycine residue.

[0053] In one embodiment, a nucleic acid molecule, in which polynucleotide encoding the polypeptide is genetically fused with polynucleotide encoding the antigen, is expressed in a host cell so that the first attachment site and the second attachment site are linked through a peptide bond. In this case, the polypeptide and the antigen are linked through a peptide bond. Relating to this embodiment, the first attachment site and/or the second attachment site may be genetically modified from the original polypeptide or antigen. For example, the first attachment site is modified from the polypeptide so that through a linker peptide including SG, GS, SGG, GGS and SGSG, the polypeptide is conjugated with the antigen.

[0054] When the polypeptide are chemically conjugated with the antigen, the first attachment site and the second attachment site may be linked through a chemical cross-linker which is a chemical compound.

[0055] Examples of the cross-linker include, but are not limited to, SMPH, Sulfo-MBS, Sulfo-EMCS, Sulfo-GMBS, Sulfo-SIAB, Sulfo-SMPB, Sulfo-SMCC, SVSB, SIA and other cross-linkers available from the Pierce Chemical Company.

[0056] In one embodiment, the particle provided by the present invention comprises a polypeptide linked to an antigen, wherein spatial distance between the N-terminal residue and C-terminal residue of the antigen is 30 .ANG. or less when the distance is determined in a crystal of the antigen or a naturally occurring protein containing the antigen or modified protein therefrom.

[0057] The antigen used for the present invention can be designed by a person skilled in the art. For example, the antigen used for the present invention may be a naturally occurring protein or a fragment thereof. Alternatively, the antigen used for the present invention may be a protein modified from a naturally occurring protein or a fragment thereof. A person skilled in the art can design the antigen so that spatial distance between the N-terminal residue and C-terminal residue of the antigen is 30 .ANG. or less when the distance is determined in a crystal of the antigen or a naturally occurring protein containing the antigen or modified protein therefrom. For example, the antigen used for the particle provided by the present invention can be designed using a free software including PyMOL (e.g. PyMOL v0.99: http:/www.pymol.org). In one embodiment, the spatial distance between the N-terminal residue and C-terminal residue of the antigen is 30 .ANG.(angstrom) or less, 20 .ANG. or less, or 10 .ANG. or less (e.g. from 5 .ANG. to 15 .ANG., from 5 .ANG. to 12 .ANG., from 5 .ANG. to 11 .ANG., from 5 .ANG. to 10 .ANG., from 5 .ANG. to 8 .ANG., from 8 .ANG. to 15 .ANG., from 8 .ANG. to 13 .ANG., from 8 .ANG. to 12 .ANG., from 8 .ANG. to 11 .ANG., from 9 .ANG. to 12 .ANG., from 9 .ANG. to 11 .ANG., from 9 .ANG. to 10 .ANG. or from 10 .ANG. to 11 .ANG.).

Chikungunya Virus Like Particle or a Venezuelan Equine Encephalitis Virus Like Particle

[0058] In one embodiment, the present invention provides a Chikungunya virus like particle or a Venezuelan equine encephalitis virus like particle comprising a Chikungunya or Venezuelan equine encephalitis virus structural polypeptide and at least one malaria antigen, wherein said Chikungunya virus structural polypeptide or said Venezuelan equine encephalitis virus structural polypeptide comprises at least one first attachment site and said at least one malaria antigen comprises at least one second attachment site, and wherein said Chikungunya or Venezuelan equine encephalitis virus structural polypeptide and said at least one antigen are linked through said at least one first and said at least one second attachment site.

[0059] In one embodiment, a spatial distance between the N-terminal residue and C-terminal residue of the malaria antigen may be 30 .ANG. or less; 25 .ANG. or less; 20 .ANG. or less; 15 .ANG. or less; 14 .ANG. or less; 13 .ANG. or less; 12 .ANG. or less; 11 .ANG. or less; 10 .ANG. or less; 9 .ANG. or less; or 8 .ANG. or less (e.g. from 5 .ANG. to 15 .ANG., from 5 .ANG. to 12 .ANG., from 5 .ANG. to 11 .ANG., from 5 .ANG. to 10 .ANG., from 5 .ANG. to 8 .ANG., from 8 .ANG. to 15 .ANG., from 8 .ANG. to 13 .ANG., from 8 .ANG. to 12 .ANG., from 8 .ANG. to 11 .ANG., from 9 .ANG. to 12 .ANG., from 9 .ANG. to 11 .ANG., from 9 .ANG. to 10 .ANG. or from 10 .ANG. to 11 .ANG.) when the distance is determined in a crystal of the malaria antigen or a naturally occurring protein containing the malaria antigen or modified protein therefrom.

[0060] In one embodiment, the malaria antigen is linked to the Chikungunya or Venezuelan equine encephalitis virus structural polypeptide by way of chemical cross-linking or as a fusion protein produced by way of genetic engineering.

[0061] A Chikungunya or Venezuelan equine encephalitis virus structural polypeptide used in the present invention may comprise a Chikungunya or Venezuelan equine encephalitis virus envelope protein and/or a capsid.

[0062] Examples of Chikungunya virus include, but are not limited to, strains of 37997 and LR2006 OPY-1.

[0063] Examples of Venezuelan equine encephalitis virus include, but are not limited to, TC-83.

[0064] Chikungunya or Venezuelan equine encephalitis virus structural polypeptide used in the present invention may naturally occurring virus structural polypeptide or modified polypeptide thereof. The modified polypeptide may be a fragment of the naturally occurring virus structural polypeptide. In one embodiment, the modified polypeptide has at least 70%, 75%, 80%, 85%, 90%, 95% or 98% amino acid sequence identity to a naturally occurring viral capsid and/or envelope protein. In one embodiment, the modified polypeptide is a mutant where at most 10% of the amino acids are deleted, substituted, and/or added based on a naturally occurring viral capsid and/or envelope protein. For example, K64A or K64N mutatation may be introduced into a capsid of Venezuelan equine encephalitis virus structural polypeptide used in the present invention.

[0065] Chikungunya or Venezuelan equine encephalitis virus structural polypeptide may consist of or comprise a capsid, E2 and E1.

[0066] Examples of Chikungunya virus structural polypeptide include, but are not limited to, Capsid-E2-E1 of Chikungunya virus Strain 37997, and Capsid-E2-E1 of Chikungunya virus LR2006 OPY-1.

[0067] Examples of Venezuelan equine encephalitis virus structural polypeptide include, but are not limited to, Capsid-E2-E1 of Venezuelan equine encephalitis virus Strain TC-83.

[0068] An exemplary Chikungunya virus structural polypeptide sequence is provided at Genbank Accession No. ABX40006.1, which is described below (SEQ ID No. 1):

TABLE-US-00003 mefiptqtfynrryqprpwtprptiqvirprprpqrqagqlaqlis avnkltmravpqqkprrnrknkkqkqkqqapqnntnqkkqppkkkp aqkkkkpgrrermcmkiendcifevkhegkvtgyaclvgdkvmkpa hvkgtidnadlaklafkrsskydlecaqipvhmksdaskfthekpe gyynwhhgavgysggrftiptgagkpgdsgrpifdnkgrvvaivig ganegartalsvvtwnkdivtkitpegaeewslaipvmcilanttf pcsqppctpccyekepeetlrmlednvmrpgyyqllqasltcsphr grrstkdnfnvykatrpylahcpdcgeghschspvalerirneatd gtlkiqvslgigiktddshdwtklrymdnhmpadaeraglevrtsa pctitgtmghfilarcpkgetltvgftdsrkishscthpfhhdppv igrekfhsrpqhgkelpcstyvqstaatteeievhmppdtpdrtlm sqqsgnvkitvngqtvrykencggsnegltttdkvinnckvdgcha avtnhkkwqynsplvprnaelgdrkgkihipfplanvtcrvpkarn ptvtygknqvimllypdhptilsyrnmgeepnyqeewvmhkkevvl tvpteglevtwgnnepykywpqlstngtahghpheiilyyyelypt mtvvvvsvatfillsmvgmaagmcmcarrrcitpyeltpgatvpfl isliccirtakaatyqeaaiylwneqqplfwlqaliplaalivlcn cirlipcccktlaflavmsvgahtvsayehvtvipntvgvpyktlv nrpgyspmvlemellsvtleptlsidyitceyktvipspyvkccgt aeckdknipdysckvftgvypfmwggaycfcdaentqlseahveks escktefasayrahtasasakirylyqgnnitvtayangdhavtvk dakfivgpmssawtpfdnkivvykgdvynmdyppfgagrpgqfgdi gsrtpeskdvyantqlvlqrpavgtvhvpysqapsgfkywlkerga slqhtapfgcgiatnpvravncavgnmpisidipeaaftrvvdaps ltdmscevpacthssdfggvaiikyaaskkgkcavhsmtnavtire aeievegnsqlqisfstalasaefrvqvcstqvhcaaechppkdhi vnypashttlgvgdisatamswvqkitggvglvvavaalilivvlc vsfsrh

[0069] Another exemplary Chikungunya virus structural polypeptide sequence is provided at Genbank Accession No. ABX40011.1, which is described below (SEQ ID No. 2):

TABLE-US-00004 mefiptqtfynrryqprpwaprptiqvirprprpqrqagqlaqlis avnkltmravpqqkprrnrknkkqrqkkqapqndpkqkkqppqkkp aqkkkkpgrrermcmkiendcifevkhegkvmgyaclvgdkvmkpa hvkgtidnadlaklafkrsskydlecaqipvhmksdaskfthekpe gyynwhhgavqysggrftiptgagkpgdsgrpifdnkgrvvaivlg ganegartalsvvtwnkdivtkitpegaeewslalpvlcllanttf pcsqppctpccyekepestlrmlednvmrpgyyqllkasltcsphr qrrstkdnfnvykatrpylahcpdcgeghschspialerirneatd gtlkiqvslqigiktddshdwtklrymdshtpadaeragllvrtsa pctitgtmghfilarcpkgetltvgftdsrkishtcthpfhheppv igrerfhsrpqhgkelpcstyvqstaataeeievhmppdtpdrtlm tqqsgnvkitvngqtvrykcncggsnegltttdkvinnckidqcha avtnhknwqynsplvprnaelgdrkgkihipfplanvtcrvpkarn ptvtygknqvtmllypdhptllsyrnmgqepnyheewvthkkevtl tvpteglevtwgnnepykywpqmstngtahghpheiilyyyelypt mtvvivsvasfvllsmvgtavgmcvcarrrcitpyeltpgatvpfl lsllccvrttkaatyyeaaaylwneqqplfwlqaliplaalivlcn clkllpcccktlaflavmsigahtvsayehvtvipntvgvpyktlv nrpgyspmvlemelqsvtleptlsldyitceyktvipspyvkccgt aeckdkslpdysckvftgvypfmwggaycfcdaentqlseahveks escktefasayrahtasasaklrvlyqgnnitvaayangdhavtvk dakfvvgpmssawtpfdnkivvykgdvynmdyppfgagrpgqfgdi qsrtpeskdvyantqlvlqrpaagtvhvpysqapsgfkywlkerga slqhtapfgcqiatnpvravncavgnipisidipdaaftrvvdaps vtdmscevpacthssdfggvaiikytaskkgkcavhsmtnavtire advevegnsqlqisfstalasaefrvqvcstqvhcaaachppkdhi vnypashttlgvqdisttamswvqkitggvglivavaalilivvlc vsfsrh.

[0070] An exemplary Venezuelan equine encephalitis virus structural polypeptide is provided at Genbank Accession No. L01443.1 (http://www.ncbi.nlm.nih.gov/nuccore/L01443.1), which is described below (SEQ ID No. 3):

TABLE-US-00005 mfpfqpmypmqpmpyrnpfaaprrpwfprtdpflamqvqeltrsma nitfkqrrdappegpsaakpkkeasqkqkgggqgkkkknqgkkkak tgppnpkaqngnkkktnkkpqkrqrmvmklesdktfpimiegking yacvvggklfrpmhvegkidndvlaalktkkaskydleyadvpqnm radtfkythekpqgyyswhhgavqyengrftvpkgvgakgdsgrpi ldnqgrvvaivaggvnegsrtalsvvmwnekgvtvkytpenceqws lvttmcilanvtfpcaqppicydrkpaetlamlsvnvdnpgydell eaavkcpgrkrrsteelfneykltrpymarcircavgschspiaie avksdghdgyvraqtssqygldssgnikgrtmrydmhgtikeiplh qvslytsrpchivdghgyfllarepagdsitmefkkdsvrhscsvp yevkfnpvgrelythppehgveqacqvyahdaqnrgayvemhlpgs evdsslvslsgssvtvtppdgtsalvececggtkisetinktkqfs qctkkeqcrayrlqndkwvynsdklpkaagatlkgklhvpflladg kctvplapepmitfgfrsyslkihpknptylitrqladephythel isepavrnftvtekgwefvwgnhppkrfwagetapgnphglphevi thyyhrypmstilglsicaaiatvsvaastwlfcrsrvacltpyrl tpnaripfclaviccartaraettwesldhlwnnnqqmfwiqllip laalivvtrllrcvccvvpflvmagaagagayehattmpsqagisy ntivnragyaplpisitptkikliptvnleyvtchyktgmdspaik ccgsqectptyrpdeqckvftgvypfmwggaycfcdtentqvskay vmksddcladhaeaykahtasvqaflnitvgehsivttvyvngetp vnfngvkitagplstawtpfdrkivqyageiynydfpeygagqpga fgdiqsrtvsssdlyantnlvlqrpkagaihvpytqapsgfeawkk dkapsikftapfgceiytnpiraencavgsiplafdipdalftrvs etptisaaectlnecvyssdfggiatvkysasksgkcavhvpsgta tikeaaveltecosatihfstanihpefrlqictsyvtckgdchpp kdhivthpqyhaqtftaaysktawtwltsllggsaviiiiglvlat ivamyvltnqkhn.

[0071] In one embodiment, a first attachment site comprises an amino group, preferably an amino group of a lysine residue. In one embodiment, the second attachment site comprises sulfhydryl group, preferably, a sulfhydryl group of a cysteine.

[0072] In one embodiment, a conjugation of more than two substances (e.g. antigen and Chikungunya or Venezuelan equine encephalitis virus structural polypeptide) through a first attachment site or a second attachment site is achieved using chemical cross linker. Examples of the cross-linker include, but are not limited to, SMPH, Sulfo-MBS, Sulfo-EMCS, Sulfo-GMBS, Sulfo-SIAB, Sulfo-SMPB, Sulfo-SMCC, SVSB, SIA and other cross-linkers available from the Pierce Chemical Company.

[0073] According to the present invention, a Chikungunya or Venezuelan equine encephalitis virus like particle comprising a Chikungunya or Venezuelan equine encephalitis virus structural polypeptide and an antigen, wherein said Chikungunya or Venezuelan equine encephalitis virus structural polypeptide and said antigen are expressed as a fusion protein can be provided.

[0074] In one embodiment, the antigen can be fused with any site of the Chikungunya or Venezuelan equine encephalitis virus structural polypeptide. For example, the antigen may be directly or indirectly linked to N- or C-terminal of the Chikungunya or Venezuelan equine encephalitis virus structural polypeptide, or the antigen may be inserted into Chikungunya or Venezuelan equine encephalitis virus structural protein.

[0075] In one embodiment, at least one antigen is inserted into E2 of Chikungunya or Venezuelan equine encephalitis virus structural protein. For example, regarding Chikungunya virus structural protein, at least one antigen is inserted between residues 519 and 520 of SEQ ID Nos.1 or 2 (i.e. between G at 519-position and Q at 520-position of SEQ ID Nos.1 or 2); between residues 530 and 531 of SEQ ID Nos.1 or 2 (i.e. between G at 530-position and S at 531-position of SEQ ID Nos.1 or 2); between residues 531 and 532 of SEQ ID Nos.1 or 2 (i.e. between S at 531-position and N at 532-position of SEQ ID Nos.1 or 2); between residues 529 and 530 of SEQ ID Nos.1 or 2(i.e. between G at 529-position and G at 530-position of SEQ ID Nos.1 or 2); or between residues 510 and 511 of SEQ ID Nos.1 or 2(i.e. between S at 510-position and G at 511-position of SEQ ID Nos.1 or 2); or between residues 511 and 512 of SEQ ID Nos.1 or 2(i.e. between G at 511-position and N at 512-position of SEQ ID Nos.1 or 2); or between residues 509 and 510 of SEQ ID Nos.1 or 2(i.e. between Q at 509-position and S at 510-position of SEQ ID Nos.1 or 2).

[0076] For example, regarding Venezuelan equine encephalitis virus structural protein, at least one antigen is inserted between residues 517 and 518 of SEQ ID No.3 (i.e. between G at 517-position and S at 518-position of SEQ ID No.3); between residues 518 and 519 of SEQ ID No.3 (i.e. between S at 518-position and S at 519-position of SEQ ID No.3); between residues 519 and 520 of SEQ ID No.3 (i.e. between S at 519-position and V at 520-position of SEQ ID No.3); between residues 515 and 516 of SEQ ID No.3(i.e. between L at 515-position and S at 516-position of SEQ ID No.3); between residues 516 and 517 of SEQ ID No.3(i.e. between S at 516-position and G at 517-position of SEQ ID No.3); between residues 536 and 537 of SEQ ID No.3(i.e. between C at 536-position and G at 537-position of SEQ ID No.3); between residues 537 and 538 of SEQ ID No.3(i.e. between G at 537-position and G at 538-position of SEQ ID No.3); between residues 538 and 539 of SEQ ID No.3(i.e. between G at 538-position and T at 539-position of SEQ ID No.3).

[0077] The fusion protein may be expressed using a conventional technique in the art. A variety of expression systems can be used for the expression of the fusion protein. For example, the fusion protein can be expressed in 293 cells, Sf9 cells or E. coli.

[0078] A polypeptide derived from Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV) may be a naturally occurring viral polypeptide or modified polypeptide thereof. In addition, a polypeptide derived from malaria antigen may be a naturally occurring polypeptide or modified polypeptide of the naturally occurring polypeptide or a fragment of the naturally occurring polypeptide or the modified peptide. The modified polypeptide may be a fragment of the naturally occurring virus structural polypeptide.

[0079] In one embodiment, the modified polypeptide derived from malaria antigen has at least 70%, 75%, 80%, 85%, 90%, 95% or 98% amino acid sequence identity to a naturally occurring polypeptide. In one embodiment, the modified peptide derived from malaria antigen is a mutant where at most 10% of the amino acids are deleted, substituted, and/or added based on a naturally occurring polypeptide derived from malaria antigen.

[0080] When a polypeptide derived from a virus is conjugated with a polypeptide derived from an antigen, a linker peptide including SG, GS, SGG, GGS SGSG and TRGGS may be used. Examples of conjugation of the polypeptide derived from a virus (referred to as "PFV" below) with the polypeptide derived from the antigen (referred to as "PFA" below) include, but not limited to: PFV-SG-PFA-GS-PFV; PFV-SG-PFA-GGS-PFV; PFV-SSG-PFA-GS-PFV; PFV-SGG-PFA-GGS-PFV; PFV-SGSG-PFA-GS-PFV; and PFA-SGG-PFA-TRGGS-PFV.

[0081] In one embodiment, the present invention provides a virus like particle comprising a fusion protein of a polypeptide derived from Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV) and a polypeptide derived from malaria antigen, wherein the virus like particle is prepared by transfecting an expression vector comprising a nucleic acid molecule corresponding to the amino acid sequence represented by SEQ ID NO. 28, 31, 34, 37, 39, 41 or 43 into a mammalian cell (e.g. 293F cell). Regarding this embodiment, modified fusion protein can be also used for a virus like particle provided by the present invention, which can be prepared by transfecting an expression vector comprising a nucleic acid molecule corresponding to the amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95% or 98% amino acid sequence identity to SEQ ID NO. 28, 31, 34, 37, 39, 41 or 43 into a mammalian cell (e.g. 293F cell).

[0082] In one embodiment, the present invention provides a virus like particle comprising or consisting of:

one or more capsid of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV); one or more E1 of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV); and one or more E2 of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV), wherein malaria antigen is inserted into E2 of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV). For example, present invention provides a virus like particle comprising or consisting of: 240 capsids of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV); 240 E1s of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV); and 240 E2s of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV), wherein malaria antigen is inserted into each of E2s of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV).

[0083] In this embodiment, the E2 into which the antigen is inserted may consist of an amino acid sequence represented by SEQ ID No.50; the E1 may consist of an amino acid sequence represented by SEQ ID No.51; and the capsid may consist of an amino acid sequence represented by SEQ ID NO.: 52; or

the E2 into which the antigen is inserted may consist of an amino acid sequence represented by SEQ ID NO.53; the E1 may consist of an amino acid sequence represented by SEQ ID NO.54; and the capsid may consist of an amino acid sequence represented by SEQ ID NO.: 55.

[0084] Further, regarding this embodiment, modified capsid of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV), modified E1 of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV) and modified E2 of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV) may be used for the virus like particle. For example, the modified capsid of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV) may have at least 70%, 75%, 80%, 85%, 90%, 95% or 98% amino acid sequence identity to the amino acid sequence represented by SEQ ID NO.: 52 or SEQ ID No. 55; the modified E1 of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV) may have at least 70%, 75%, 80%, 85%, 90%, 95% or 98% amino acid sequence identity to the amino acid sequence represented by SEQ ID NO.: 51 or SEQ ID No. 54; and/or the modified E2 of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV) may have at least 70%, 75%, 80%, 85%, 90%, 95% or 98% amino acid sequence identity to the amino acid sequence represented by SEQ ID NO.: 50 or SEQ ID No. 53.

[0085] Also, the modified capsid, E1 or E2 may be a mutant where at most 10% of the amino acids are deleted, substituted, and/or added based on the capsid consisting of an amino acid sequence represented by SEQ ID NO.: 52 or SEQ ID No. 55; E1 consisting of an amino acid sequence represented by SEQ ID NO.: 51 or SEQ ID No. 54; and/or E2 consisting of an amino acid sequence represented by SEQ ID NO.: 50 or SEQ ID No. 53.

(2) Nucleotide, Vector, Host cell

[0086] In the second aspect, the present invention provides a nucleic acid molecule which is designed for expression of a particle as provided in the first aspect of the present invention.

[0087] In one embodiment, the present invention provides a nucleic acid molecule comprising a nucleotide sequence that encodes the Chikungunya or Venezuelan equine encephalitis virus like particle as described above.

[0088] Examples of the nucleotide sequence that encodes the Chikungunya or Venezuelan equine encephalitis virus like particle include, but are not limited to, a nucleotide sequence encoding envelope of Chikungunya virus Strain 37997, a nucleotide sequence encoding Capsid-envelope of Chikungunya virus Strain 37997, a nucleotide sequence encoding envelope of Chikungunya virus Strain LR2006 OPY-1, a nucleotide sequence encoding Capsid-envelope of Chikungunya virus LR2006 OPY-1, a nucleotide sequence encoding envelope of Venezuelan equine encephalitis virus Strain TC-83 and a nucleotide sequence encoding Capsid-envelope of Venezuelan equine encephalitis virus TC-83.

[0089] Regarding Chikungunya virus, an exemplary nucleotide sequence that encodes envelope is described below (SEQ ID No.4):

TABLE-US-00006 Atgagcctcgccctcccggtcttgtgcctgttggcaaacactacat tcccctgctctcagccgccttgcacaccctgctgctacgaaaagga accggaaagcaccttgcgcatgcttgaggacaacgtgatgagaccc ggatactaccagctactaaaagcatcgctgacttgctctccccacc gccaaagacgcagtactaaggacaattttaatgtctataaagccac aagaccatatctagctcattgtcctgactgcggagaagggcattcg tgccacagccctatcgcattggagcgcatcagaaatgaagcaacgg acggaacgctgaaaatccaggtctctttgcagatcgggataaagac agatgacagccacgattggaccaagctgcgctatatggatagccat acgccagcggacgcggagcgagccggattgcttgtaaggacttcag caccgtgcacgatcaccgggaccatgggacactttattctcgcccg atgcccgaaaggagagacgctgacagtgggatttacggacagcaga aagatcagccacacatgcacacacccgttccatcatgaaccacctg tgataggtagggagaggttccactctcgaccacaacatggtaaaga gttaccttgcagcacgtacgtgcagagcaccgctgccactgctgag gagatagaggtgcatatgcccccagatactcctgaccgcacgctga tgacgcagcagtctggcaacgtgaagatcacagttaatgggcagac ggtgcggtacaagtgcaactgcggtggctcaaacgagggactgaca accacagacaaagtgatcaataactgcaaaattgatcagtgccatg ctgcagtcactaatcacaagaattggcaatacaactcccctttagt cccgcgcaacgctgaactcggggaccgtaaaggaaagatccacatc ccattcccattggcaaacgtgacttgcagagtgccaaaagcaagaa accctacagtaacttacggaaaaaaccaagtcaccatgctgctgta tcctgaccatccgacactcttgtcttaccgtaacatgggacaggaa ccaaattaccacgaggagtgggtgacacacaagaaggaggttacct tgaccgtgcctactgagggtctggaggtcacttggggcaacaacga accatacaagtactggccgcagatgtctacgaacggtactgctcat ggtcacccacatgagataatcttgtactattatgagctgtacccca ctatgactgtagtcattgtgtcggtggcctcgttcgtgcttctgtc gatggtgggcacagcagtgggaatgtgtgtgtgcgcacggcgcaga tgcattacaccatatgaattaacaccaggagccactgttcccttcc tgctcagcctgctatgctgcgtcagaacgaccaaggcggccacata ttacgaggctgcggcatatctatggaacgaacagcagcccctgttc tggttgcaggctcttatcccgctggccgccttgatcgtcctgtgca actgtgcaactgtctgaaactcttgccatgctgctgtaagaccctg gcttttttagccgtaatgagcatcggtgcccacactgtgagcgcgt acgaacacgtaacagtgatcccgaacacggtgggagtaccgtataa gactcttgtcaacagaccgggttacagccccatggtgttggagatg gagctacaatcagtcaccttggaaccaacactgtcacttgactaca tcacgtgcgagtacaaaactgtcatcccctccccgtacgtgaagtg ctgtggtacagcagagtgcaaggacaagagcctaccagactacagc tgcaaggtctttactggagtctacccatttatgtggggcggcgcct actgcttttgcgacgccgaaaatacgcaattgagcgaggcacatgt agagaaatctgaatcttgcaaaacagagtttgcatcggcctacaga gcccacaccgcatcggcgtcggcgaagctccgcgtcctttaccaag gaaacaacattaccgtagctgcctacgctaacggtgaccatgccgt cacagtaaaggacgccaagtttgtcgtgggcccaatgtcctccgcc tggacaccttttgacaacaaaatcgtggtgtacaaaggcgacgtct acaacatggactacccaccttttggcgcaggaagaccaggacaatt tggtgacattcaaagtcgtacaccggaaagtaaagacgtttatgcc aacactcagttggtactacagaggccagcagcaggcacggtacatg taccatactctcaggcaccatctggcttcaagtattggctgaagga acgaggagcatcgctacagcacacggcaccgttcggttgccagatt gcgacaaacccggtaagagctgtaaattgcgctgtggggaacatac caatttccatcgacataccggatgcggcctttactagggttgtcga tgcaccctctgtaacggacatgtcatgcgaagtaccagcctgcact cactcctccgactttgggggcgtcgccatcatcaaatacacagcta gcaagaaaggtaaatgtgcagtacattcgatgaccaacgccgttac cattcgagaagccgacgtagaagtagaggggaactcccagctgcaa atatccttctcaacagccctggcaagcgccgagtttcgcgtgcaag tgtgctccacacaagtacactgcgcagccgcatgccaccctccaaa ggaccacatagtcaattacccagcatcacacaccacccttggggtc caggatatatccacaacggcaatgtcttgggtgcagaagattacgg gaggagtaggattaattgttgctgttgctgccttaattttaattgt ggtgctatgcgtgtcgtttagcaggcac

[0090] Regarding Chikungunya virus, another exemplary nucleotide sequence that encodes envelope is described below (SEQ ID No. 5):

TABLE-US-00007 Atgagtcttgccatcccagttatgtgcctgttggcaaacaccacgtt cccctgctcccagcccccttgcacgccctgctgctacgaaaaggaac cggaggaaaccctacgcatgcttgaggacaacgtcatgagacctggg tactatcagctgctacaagcatccttaacatgttctccccaccgcca gcgacgcagcaccaaggacaacttcaatgtctataaagccacaagac catacttagctcactgtcccgactgtggagaagggcactcgtgccat agtcccgtagcactagaacgcatcagaaatgaagcgacagacgggac gctgaaaatccaggtctccttgcaaatcggaataaagacggatgaca gccacgattggaccaagctgcgttatatggacaaccacatgccagca gacgcagagagggcggggctatttgtaagaacatcagcaccgtgtac gattactggaacaatgggacacttcatcctggcccgatgtccaaaag gggaaactctgacggtgggattcactgacagtaggaagattagtcac tcatgtacgcacccatttcaccacgaccctcctgtgataggtcggga aaaattccattcccgaccgcagcacggtaaagagctaccttgcagca cgtacgtgcagagcaccgccgcaactaccgaggagatagaggtacac atgcccccagacacccctgatcgcacattaatgtcacaacagtccgg caacgtaaagatcacagtcaatggccagacggtgcggtacaagtgta attgcggtggctcaaatgaaggactaacaactacagacaaagtgatt aataactgcaaggttgatcaatgtcatgccgcggtcaccaatcacaa aaagtggcagtataactcccctctggtcccgcgtaatgctgaacttg gggaccgaaaaggaaaaattcacatcccgtttccgctggcaaatgta acatgcagggtgcctaaagcaaggaaccccaccgtgacgtacgggaa aaaccaagtcatcatgctactgtatcctgaccacccaacactcctgt cctaccggaatatgggagaagaaccaaactatcaagaagagtgggtg atgcataagaaggaagtcgtgctaaccgtgccgactgaagggctcga ggtcacgtggggcaacaacgagccgtataagtattggccgcagttat ctacaaacggtacagcccatggccacccgcatgagataattctgtat tattatgagctgtaccccactatgactgtagtagttgtgtcagtggc cacgttcatactcctgtcgatggtgggtatggcagcggggatgtgca tgtgtgcacgacgcagatgcatcacaccgtatgaactgacaccagga gctaccgtccctttcctgcttagcctaatatgctgcatcagaacagc taaagcggccacataccaagaggctgcgatatacctgtggaacgagc agcaacctttgttttggctacaagcccttattccgctggcagccctg attgttctatgcaactgtctgagactcttaccatgctgctgtaaaac gttggcttttttagccgtaatgagcgtcggtgcccacactgtgagcg cgtacgaacacgtaacagtgatcccgaacacggtgggagtaccgtat aagactctagtcaatagacctggctacagccccatggtattggagat ggaactactgtcagtcactttggagccaacactatcgcttgattaca tcacgtgcgagtacaaaaccgtcatcccgtctccgtacgtgaagtgc tgcggtacagcagagtgcaaggacaaaaacctacctgactacagctg taaggtcttcaccggcgtctacccatttatgtggggcggcgcctact gcttctgcgacgctgaaaacacgcagttgagcgaagcacacgtggag aagtccgaatcatgcaaaacagaatttgcatcagcatacagggctca taccgcatctgcatcagctaagctccgcgtcctttaccaaggaaata acatcactgtaactgcctatgcaaacggcgaccatgccgtcacagtt aaggacgccaaattcattgtggggccaatgtcttcagcctggacacc tttcgacaacaaaattgtggtgtacaaaggtgacgtctataacatgg actacccgccctttggcgcaggaagaccaggacaatttggcgatatc caaagtcgcacacctgagagtaaagacgtctatgctaatacacaact ggtactgcagagaccggctgtgggtacggtacacgtgccatactctc aggcaccatctggctttaagtattggctaaaagaacgcggggcgtcg ctgcagcacacagcaccatttggctgccaaatagcaacaaacccggt aagagcggtgaactgcgccgtagggaacatgcccatctccatcgaca taccggaagcggccttcactagggtcgtcgacgcgccctctttaacg gacatgtcgtgcgaggtaccagcctcgacccattcctcagactttgg gggcgtcgccattattaaatatgcagccagcaagaaaggcaagtgtg cggtgcattcgatgactaacgccgtcactattcgggaagctgagata gaagttgaagggaattctcagctgcaaatctctttctcgacggcctt agccagcgccgaattccgcgtacaagtctgttctacacaagtacact gtgcagccgagtgccaccccccgaaggaccacatagtcaactacccg gcgtcacataccaccctcggggtccaggacatctccgctacggcgat gtcatgggtgcagaagatcacgggaggtgtgggactggttgttgctg ttgccgcactgattctaatcgtggtgctatgcgtgtcgttcagcagg cac

[0091] Regarding Chikungunya virus, an exemplary nucleotide sequence that encodes a Capsid-envelope is described below (SEQ ID No. 6):

TABLE-US-00008 atggagttcatcccgacgcaaactttctataacagaaggtaccaacc ccgaccctgggccccacgccctacaattcaagtaattagacctagac cacgtccacagaggcaggctgggcaactcgcccagctgatctccgca gtcaacaaattgaccatgcgcgcggtacctcaacagaagcctcgcag aaatcggaaaaacaagaagcaaaggcagaagaagcaggcgccgcaaa acgacccaaagcaaaagaagcaaccaccacaaaagaagccggctcaa aagaagaagaaaccaggccgtagggagagaatgtgcatgaaaattga aaatgattgcatcttcgaagtcaagcatgaaggcaaagtgatgggct acgcatgcctggtgggggataaagtaatgaaaccagcacatgtgaag ggaactatcgacaataccgatctggctaaactggcctttaagcggtc gtctaaatacgatcttgaatgtgcacagataccggtgcacatgaagt ctgatgcctcgaagtttacccacgagaaacccgaggggtactataac tggcatcacggagcagtgcagtattcaggaggccggttcactatccc gacgggtgcaggcaagccgggagacagcggcagaccgatcttcgaca acaaaggacgggtggtggccatcgtcctaggaggggccaacgaaggt gcccgcacggccctotccgtggtgacgtggaacaaagacatcgtcac aaaaattacccctgagggagccgaagagtggagcctcgccctcccgg tcttgtgcctgttggcaaacactacattcccctgctctcagccgcct tgcacaccctgctgctacgaaaaggaaccggaaagcaccttgcgcat gcttgaggacaacgtgatgagacccggatactaccagctactaaaag catcgctgacttgctctccccaccgccaaagacgcagtactaaggac aattttaatgtctataaagccacaagaccatatctagctcattgtcc tgactgcggagaagggcattcgtgccacagccctatcgcattggagc gcatcagaaatgaagcaacggacggaacgctgaaaatccaggtctct ttgcagatcgggataaagacagatgacagccacgattggaccaagct gcgctatatggatagccatacgccagcggacgcggagcgagccggat tgcttgtaaggacttcagcaccgtgcacgatcaccgggaccatggga cactttattctcgcccgatgcccgaaaggagagacgctgacagtggg atttacggacagcagaaagatcagccacacatgcacacacccgttcc atcatgaaccacctgtgataggtagggagaggttccactctcgacca caacatggtaaagagttaccttgcagcacgtacgtgcagagcaccgc tgccactgctgaggagatagaggtgcatatgcccccagatactcctg accgcacgctgatgacgcagcagtctggcaacgtgaagatcacagtt aatgggcagacggtgcggtacaagtgcaactgcggtggctcaaacga gggactgacaaccacagacaaagtgatcaataactgcaaaattgatc agtgccatgctgcagtcactaatcacaagaattggcaatacaactcc cctttagtcccgcgcaacgctgaactcggggaccgtaaaggaaagat ccacatcccattcccattggcaaacgtgacttgcagagtgccaaaag caagaaaccctacagtaacttacggaaaaaaccaagtcaccatgctg ctgtatcctgaccatccgacactcttgtcttaccgtaacatgggaca ggaaccaaattaccacgaggagtgggtgacacacaagaaggaggtta ccttgaccgtgcctactgagggtctggaggtcacttggggcaacaac gaaccatacaagtactggccgcagatgtctacgaacggtactgctca tggtcacccacatgagataatcttgtactattatgagctgtacccca ctatgactgtagtcattgtgtcggtggcctcgttcgtgcttctgtcg atggtgggcacagcagtgggaatgtgtgtgtgcgcacggcgcagatg cattacaccatatgaattaacaccaggagccactgttcccttcctgc tcagcctgctatgctgcgtcagaacgaccaaggcggccacatattac gaggctgcggcatatctatggaacgaacagcagcccctgttctggtt gcaggctcttatcccgctggccgccttgatcgtcctgtgcaactgtc tgaaactcttgccatgctgctgtaagaccctggcttttttagccgta atgagcatcggtgcccacactgtgagcgcgtacgaacacgtaacagt gatcccgaacacggtgggagtaccgtataagactcttgtcaacagac cgggttacagccccatggtgttggagatggagctacaatcagtcacc ttggaaccaacactgtcacttgactacatcacgtgcgagtacaaaac tgtcatcccctccccgtacgtgaagtgctgtggtacagcagagtgca aggacaagagcctaccagactacagctgcaaggtctttactggagtc tacccatttatgtggggcggcgcctactgcttttgcgacgccgaaaa tacgcaattgagcgaggcacatgtagagaaatctgaatcttgcaaaa cagagtttgcatcggcctacagagcccacaccgcatcggcgtcggcg aagctccgcgtcctttaccaaggaaacaacattaccgtagctgccta cgctaacggtgaccatgccgtcacagtaaaggacgccaagtttgtcg tgggcccaatgtcctccgcctggacaccttttgacaacaaaatcgtg gtgtacaaaggcgacgtctacaacatggactacccaccttttggcgc aggaagaccaggacaatttggtgacattcaaagtcgtacaccggaaa gtaaagacgtttatgccaacactcagttggtactacagaggccagca gcaggcacggtacatgtaccatactctcaggcaccatctggcttcaa gtattggctgaaggaacgaggagcatcgctacagcacacggcaccgt tcggttgccagattgcgacaaacccggtaagagctgtaaattgcgct gtggggaacataccaatttccatcgacataccggatgcggcctttac tagggttgtcgatgcaccctctgtaacggacatgtcatgcgaagtac cagcctgcactcactcctccgactttgggggcgtcgccatcatcaaa tacacagctagcaagaaaggtaaatgtgcagtacattcgatgaccaa cgccgttaccattcgagaagccgacgtagaagtagaggggaactccc agctgcaaatatccttctcaacagccctggcaagcgccgagtttcgc gtgcaagtgtgctccacacaagtacactgcgcagccgcatgccaccc tccaaaggaccacatagtcaattacccagcatcacacaccacccttg gggtccaggatatatccacaacggcaatgtcttgggtgcagaagatt acgggaggagtaggattaattgttgctgttgctgccttaattttaat tgtggtgctatgcgtgtcgtttagcaggcactaa.

[0092] Regarding Chikungunya virus, another exemplary nucleotide sequence that encodes a Capsid-envelope is described below (SEQ ID No. 7):

TABLE-US-00009 atggagttcatcccaacccaaactttttacaataggaggtaccagcc tcgaccctggactccgcgccctactatccaagtcatcaggcccagac cgcgccctcagaggcaagctgggcaacttgcccagctgatctcagca gttaataaactgacaatgcgcgcggtaccacaacagaagccacgcag gaatcggaagaataagaagcaaaagcaaaaacaacaggcgccacaaa acaacacaaatcaaaagaagcagccacctaaaaagaaaccggctcaa aagaaaaagaagccgggccgcagagagaggatgtgcatgaaaatcga aaatgattgtattttcgaagtcaagcacgaaggtaaggtaacaggtt acgcgtgcctggtgggggacaaagtaatgaaaccagcacacgtaaag gggaccatcgataacgcggacctggccaaactggcctttaagcggtc atctaagtatgaccttgaatgcgcgcagatacccgtgcacatgaagt ccgacgcttcgaagttcacccatgagaaaccggaggggtactacaac tggcaccacggagcagtacagtactcaggaggccggttcaccatccc tacaggtgctggcaaaccaggggacagcggcagaccgatcttcgaca acaagggacgcgtggtggccatagtcttaggaggagctaatgaagga gcccgtacagccctctcggtggtgacctggaataaagacattgtcac taaaatcacccccgagggggccgaagagtggagtcttgccatcccag ttatgtgcctgttggcaaacaccacgttcccctgctcccagccccct tgcacgccctgctgctacgaaaaggaaccggaggaaaccctacgcat gcttgaggacaacgtcatgagacctgggtactatcagctgctacaag catccttaacatgttctccccaccgccagcgacgcagcaccaaggac aacttcaatgtctataaagccacaagaccatacttagctcactgtcc cgactgtggagaagggcactcgtgccatagtcccgtagcactagaac gcatcagaaatgaagcgacagacgggacgctgaaaatccaggtctcc ttgcaaatcggaataaagacggatgacagccacgattggaccaagct gcgttatatggacaaccacatgccagcagacgcagagagggcggggc tatttgtaagaacatcagcaccgtgtacgattactggaacaatggga cacttcatcctggcccgatgtccaaaaggggaaactctgacggtggg attcactgacagtaggaagattagtcactcatgtacgcacccatttc accacgaccctcctgtgataggtcgggaaaaattccattcccgaccg cagcacggtaaagagctaccttgcagcacgtacgtgcagagcaccgc cgcaactaccgaggagatagaggtacacatgcccccagacacccctg atcgcacattaatgtcacaacagtccggcaacgtaaagatcacagtc aatggccagacggtgcggtacaagtgtaattgcggtggctcaaatga aggactaacaactacagacaaagtgattaataactgcaaggttgatc aatgtcatgccgcggtcaccaatcacaaaaagtggcagtataactcc cctctggtcccgcgtaatgctgaacttggggaccgaaaaggaaaaat tcacatcccgtttccgctggcaaatgtaacatgcagggtgcctaaag caaggaaccccaccgtgacgtacgggaaaaaccaagtcatcatgcta ctgtatcctgaccacccaacactcctgtcctaccggaatatgggaga agaaccaaactatcaagaagagtgggtgatgcataagaaggaagtcg tgctaaccgtgccgactgaagggctcgaggtcacgtggggcaacaac gagccgtataagtattggccgcagttatctacaaacggtacagccca tggccacccgcatgagataattctgtattattatgagctgtacccca ctatgactgtagtagttgtgtcagtggccacgttcatactcctgtcg atggtgggtatggcagcggggatgtgcatgtgtgcacgacgcagatg catcacaccgtatgaactgacaccaggagctaccgtccctttcctgc ttagcctaatatgctgcatcagaacagctaaagcggccacataccaa gaggctgcgatatacctgtggaacgagcagcaacctttgttttggct acaagcccttattccgctggcagccctgattgttctatgcaactgtc tgagactcttaccatgctgctgtaaaacgttggcttttttagccgta atgagcgtcggtgcccacactgtgagcgcgtacgaacacgtaacagt gatcccgaacacggtgggagtaccgtataagactctagtcaatagac ctggctacagccccatggtattggagatggaactactgtcagtcact ttggagccaacactatcgcttgattacatcacgtgcgagtacaaaac cgtcatcccgtctccgtacgtgaagtgctgcggtacagcagagtgca aggacaaaaacctacctgactacagctgtaaggtcttcaccggcgtc tacccatttatgtggggcggcgcctactgcttctgcgacgctgaaaa cacgcagttgagcgaagcacacgtggagaagtccgaatcatgcaaaa cagaatttqcatcagcatacagggctcataccgcatctgcatcagct aagctccgcgtcctttaccaaggaaataacatcactgtaactgccta tgcaaacggcgaccatgccgtcacagttaaggacgccaaattcattg tggggccaatgtcttcagcctggacacctttcgacaacaaaattgtg gtgtacaaaggtgacgtctataacatggactacccgccctttggcgc aggaagaccaggacaatttggcgatatccaaagtcgcacacctgaga gtaaagacgtctatgctaatacacaactggtactgcagagaccggct gtgggtacggtacacgtgccatactctcaggcaccatctggctttaa gtattggctaaaagaacgcggggcgtcgctgcagcacacagcaccat ttggctgccaaatagcaacaaacccggtaagagcggtgaactgcgcc gtagggaacatgcccatctccatcgacataccggaagcggccttcac tagggtcgtcgacgcgccctctttaacggacatgtcgtgcgaggtac cagcctgcacccattcctcagactttgggggcgtcgccattattaaa tatgcagccagcaagaaaggcaagtgtgcggtgcattcgatgactaa cgccgtcactattcgggaagctgagatagaagttgaagggaattctc agctgcaaatctctttctcgacggccttagccagcgccgaattccgc gtacaagtctgttctacacaagtacactgtgcagccgagtgccaccc cccgaaggaccacatagtcaactacccggcgtcacataccaccctcg gggtccaggacatctccgctacggcgatgtcatgggtgcagaagatc acgggaggtgtgggactggttgttgctgttgccgcactgattctaat cgtggtgctatgcgtgtcgttcagcaggcactaa.

[0093] In one embodiment, the present invention provides a vector comprising the nucleic acid molecule as described above, wherein the vector optionally comprises an expression control sequence operably linked to the nucleic acid molecule.

[0094] Examples of an expression control sequence include, but are not limited to, promoter such as CMV promoter, phage lambda PL promoter, the E. coli lac, phoA and tac promoters, the SV40 early and late promoters, and promoters of retroviral LTRs.

[0095] In this embodiment, the vector comprising an expression control sequence operably linked to the nucleic acid molecule as described above can be used as an expression vector for preparing the particle provided by the first aspect of the present invention.

[0096] The expression vectors can be prepared by a person skilled in the art based on WO/2012/006180, the entire contents of which are incorporated by reference herein.

[0097] Examples of vectors which can be used for expressing a virus like particle comprising a fusion protein of a polypeptide derived from Chikungunya virus (CHIKV) and a polypeptide of antigen include a vector shown in VLP_CHI 512 vector (SEQ ID No. 8) containing CHIKV VLP polynucleotide (SEQ ID No. 13; corresponding amino acid sequence represented by SEQ ID No.14); and VLP_CHI 532 vector (SEQ ID No.: 9) containing CHIKV VLP polynucleotide (SEQ ID No. 15; corresponding amino acid sequence represented by SEQ ID No. 16)

[0098] The expression vectors can be prepared by a person skilled in the art based on US2012/0003266, the entire contents of which are incorporated by reference herein.

[0099] Examples of vectors which can be used for expressing a virus like particle comprising a fusion protein of a polypeptide derived from Venezuelan equine encephalitis virus (VEEV) and a polypeptide of antigen include a vector shown in VLP_VEEV VLP 518 vector (SEQ ID No. 10) containing VEEV VLP polynucleotide (SEQ ID No. 17; corresponding amino acid sequence represented by SEQ ID No.18); VLP_VEEV VLP 519 vector (SEQ ID No.11) containing VEEV VLP polynucleotide (SEQ ID No. 19; corresponding amino acid sequence represented by SEQ ID No.20); and VLP_VEEV VLP 538 vector (SEQ ID No.: 12) containing VEEV VLP polynucleotide (SEQ ID No. 21; corresponding amino acid sequence represented by SEQ ID No. 22)

[0100] In one embodiment, the present invention provides a nucleic acid molecule which is designed for expression of a virus like particle comprising a fusion protein of a polypeptide derived from Chikungunya virus (CHIKV) or Venezuela equine encephalitis virus (VEEV) and a polypeptide derived from malaria antigen, which consists of a nucleotide sequence represented by SEQ ID Nos.26-27, 29-30, 32-33, 35-36, 38, 40 or 42.

[0101] In one embodiment, the present invention provides a nucleic acid molecule which is modified from the nucleic acid molecule having a nucleotide sequence represented by any one of SEQ ID Nos.26-27, 29-30, 32-33 or 35-36, 38, 40 or 42. The modified nucleic acid molecule may have at least 70%, 75%, 80%, 85%, 90%, 95% or 98% nucleotide sequence identity to the nucleic acid molecule having a nucleotide sequence represented by any one of SEQ ID Nos.26-27, 29-30, 32-33, 35-36, 38, 40 or 42. Also, the modified nucleic acid molecule may be a mutant where at most 10% of the amino acids are deleted, substituted, and/or added based on the nucleic acid molecule having a nucleotide sequence represented by any one of SEQ ID Nos.26-27, 29-30, 32-33, 35-36, 38, 40 or 42.

(3) Composition or Vaccine

[0102] In the third aspect, the present invention provides a composition or vaccine comprising the particle provided in the first aspect of the present invention and/or the nucleic acid molecule provided in the second aspect of the present invention.

[0103] In one embodiment, the present invention provides a composition or vaccine comprising the Alphavirus or Flavivirus virus like particle (e.g. Chikungunya virus like particle or Venezuelan equine encephalitis virus like particle) as described above or the nucleic acid molecule as described above. The content of the Alphavirus or Flavivirus virus like particle and the content of the nucleic acid molecule may be 0.00001-1 w/w %.

[0104] Dosage amount of the particle provided in the first aspect of the present invention (e.g. CHIKV VLP or VEEV VLP) may be 1-500 .mu.g/day.

[0105] One or more malaria antigens may be used for one composition or one vaccine provided by the third aspect of the present invention.

[0106] The composition or vaccine may further comprise a pharmaceutical acceptable carrier and/or adjuvant. Examples of adjuvant include, but are not limited to, aluminium salts, sodium hydroxide, Freund's complete adjuvant, Freund's incomplete adjuvant and Ribi solution (Sigma Adjuvant system, Sigma-Aldrich). The composition or vaccine provided in the third aspect of the present invention may contain buffering agent such as dibasic sodium phosphate hydrate, sodium dihydrogen phosphate and sodium chloride; and preserving agent such as thimerosal. In one embodiment, the composition or vaccine is an aqueous solution containing 0.001-1 w/w % of the particle provided in the first aspect of the present invention (e.g. CHIKV VLP or VEEV VLP), 1-10w/w % of buffering agent, 0.01-1w/w % of adjuvant and 0.00001-0.001w/w % of preserving agent.

[0107] A skilled person can prepare the pharmaceutical composition and vaccine using conventional technique. For example, the particle provided in the first aspect of the present invention is mixed with buffer solution having physiological pH (e.g. pH 5-9, pH7) to prepare the pharmaceutical composition and vaccine provided in the third aspect of the present invention.

[0108] The pharmaceutical composition of the present invention may contain a single active ingredient or a combination of two or more active ingredients, as far as they are not contrary to the objects of the present invention. For example, cytokines including chemokines, anti-body of cytokines such as anti TNF antibody (e.g. infliximab, adalimumab), anti-VEGF antibody (e.g. bevacizumab and ranibizumab), cytokine receptor antagonist such as anti HER2 antibody (e.g. Trastuzumab), anti EGF receptor antibody (e.g. Cetuximab), anti VEGF aptamer (e.g. Pegaptanib) and immunomodulator such as cyclosporine, tacrolimus, ubenimex may be used for the combination therapy.

[0109] In a combination of plural active ingredients, their respective contents may be suitably increased or decreased in consideration of their therapeutic effects and safety.

[0110] The term "combination" used herein means two or more active ingredient are administered to a patient simultaneously in the form of a single entity or dosage, or are both administered to a patient as separate entities either simultaneously or sequentially with no specific time limits, wherein such administration provides therapeutically effective levels of the two components in the body, preferably at the same time.

[0111] In one embodiment, the composition is a vaccine composition including a DNA vaccine. In one embodiment, the DNA vaccine provided by the present invention comprises CpG containing oligonucleotide.

[0112] The composition or vaccine provided in the third aspect of the present invention can be administered one or more times. When the composition or vaccine provided in the third aspect of the present invention is administered more than one time, different particle provided in the first aspect of the present invention (e.g. CHIKV VLP or VEEV VLP) may be used for each of the administration. In one embodiment, combination of immunization using CHIKV VLP provided in the first aspect of the invention and immunization using VEEV VLP provided in the first aspect of the invention is employed. For example, CHIKV VLP provided in the first aspect of the present invention may be used for the 1st immunization and VEEV VLP provided in the first aspect of the present invention may be used for the 2nd immunization, or VEEV VLP provided in the first aspect of the present invention may be used for the 1st immunization and CHIKV VLP provided in the first aspect of the present invention may be used for the 2nd immunization.

[0113] A skilled person can determine timing of immunization using the composition or vaccine provided in the third aspect of the present invention. For example, 2nd immunization is performed 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 weeks after 1st immunization.

[0114] In one embodiment, the present invention provides a kit comprising

(a) a vaccine composition comprising the particle provided in the first aspect of the present invention; and (b) another vaccine composition comprising the particle provided in the first aspect of the present invention, wherein the particle contained in (a) is a virus like particle which is different from the particle contained in (b). In this embodiment, the particle contained in (a) may be Chikungunya virus like particle and the particle contained in (b) may be Venezuelan equine encephalitis virus like particle.

[0115] In one embodiment, the present invention provides a kit comprising

(a) a vaccine composition comprising the particle provided in the first aspect of the present invention; and (b) another vaccine composition comprising the particle provided in the first aspect of the present invention, (c) one or more vaccine composition, each of which comprises the particle provided in the first aspect of the present invention, wherein (a) is used for priming immunization and (b) and (c) are used for boosting immunization; and the particle contained in (a) is a virus like particle which is different from the particle contained in (b); and the particle contained in (c) is different from the particle contained in (a) and (b), or the same as the particle contained in (a) or (b).

[0116] The respective vaccine compositions contained in the above-described kit may be administered simultaneously, separately or sequentially.

[0117] The Alphavirus or Flavivirus virus like particle (e.g. Chikungunya virus or Venezuelan equine encephalitis virus) provided in the first aspect of the present invention or the nucleic acid molecule provided by the second aspect of the invention can be used for the composition and vaccine provided in the third aspect of the present invention.

[0118] For example, Chikungunya or Venezuelan equine encephalitis virus like particle comprising or consisting of:

one or more (e.g. 240) capsid of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV); one or more (e.g. 240) E1 of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV); and one or more (e.g. 240) E2 of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV), wherein malaria antigen is inserted into E2 of Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV) may be used for preparing the composition or vaccine provided in the third aspect of the present invention. The E2 into which the antigen is inserted may consist of an amino acid sequence represented by SEQ ID No.50; the E1 may consist of an amino acid sequence represented by SEQ ID No.51; and the capsid may consist of an amino acid sequence represented by SEQ ID NO.: 52; or the E2 into which the antigen is inserted may consist of an amino acid sequence represented by SEQ ID NO.53; the E1 may consist of an amino acid sequence represented by SEQ ID NO.54; and the capsid may consist of an amino acid sequence represented by SEQ ID NO.: 55.

[0119] The composition or vaccine provided in the third aspect of the present invention can be administered to a mammal (e.g. human) intramuscularly (i.m.), intracutaneously (i.c.), subcutaneously (s.c.), intradermally (i.d.) or intraperitoneally (i.p.).

[0120] The composition or vaccine provided in the third aspect of the present invention may be used for treating or preventing malaria.

[0121] Thus, use of the Alphavirus or Flavivirus (e.g. Chikungunya virus or Venezuelan equine encephalitis virus) virus like particle provided in the first aspect of the present invention or the nucleic acid molecule provided by the second aspect of the invention for manufacturing a pharmaceutical composition or vaccine for treating or preventing malaria is also provided by the present invention.

(4) Method of Producing an Antibody, Method of Immunomodulation, Method of Treating Malaria, Method of Inducing and/or Enhancing Immune Response Against a Malaria Antigen in a Mammal, Method of Passive Immunization, Method of Presenting an Antigen on Macrophage, and Method for Producing a Particle

[0122] In the fourth aspect, the present invention provides a method of producing an antibody, comprising contacting the particle provided in the first aspect of the present invention and/or the nucleic acid molecule provided in the second aspect of the present invention to a mammal.

[0123] The antibody produced in the fourth aspect of the present invention may be humanized using a conventional technique. Thus, in one embodiment, the method provided in the fourth aspect of the invention further comprises a step of humanizing non-human mammal produced antibody.

[0124] The particle provided in the first aspect of the present invention and/or the nucleic acid molecule provided in the second aspect of the present invention may be administered directly into the patient, into the affected organ or systemically, or applied ex vivo to cells derived from the patient or a human cell line which are subsequently administered to the patient, or used in vitro to select a subpopulation from immune cells such as B-cell and T-cell derived from the patient, which are then re-administered to the patient.

[0125] According to the present invention, the virus like particle can be applied for the immune therapy.

[0126] In the fifth aspect, the present invention provides a method of immunomodulation, a method of treating malaria, a method of inducing and/or enhancing immune response against a malaria antigen in a mammal comprising administering the composition provided in the third aspect of the present invention to a mammal.

[0127] In sixth aspect, the present invention provides a method of passive immunization against a malaria-causing pathogen, comprising administering the antibody provided in the fourth aspect of the present invention to a mammal.

[0128] In seventh aspect, the present invention provides a method of presenting a malaria antigen on macrophage, comprising contacting the particle provided in the first aspect of the present invention and/or the nucleic acid molecule provided in the second aspect of the present invention to a mammal.

[0129] In eighth aspect, the present invention provides a method for producing the particle provided in the first aspect of the present invention, comprising preparing a vector designed for expression of said particle; culturing a cell which is transfected with said vector to express said particle; and recovering said particle.

[0130] Examples of mammal include, but are not limited to, a human.

[0131] In one embodiment, the present invention provides a method of producing an antibody against malaria antigen, comprising contacting the Chikungunya or Venezuelan equine encephalitis virus like particle as described above and/or the nucleic acid molecule as described above to a mammal. The produced antibody may be an antibody which can specifically bind to a malaria antigen comprised in the Chikungunya or Venezuelan equine encephalitis virus like particle or a malaria antigen encoded by the nucleic acid molecule. The method of producing an antibody provided by the present invention may be a useful for producing a monoclonal or polyclonal antibody against a malaria antigen.

[0132] In one embodiment, an antibody against malaria antigen obtained by the method of producing an antibody according to the present invention is used for passive immunization. The method of passive immunization may comprise administering the obtained antibody to a mammal.

[0133] In one preferred embodiment, the immunomodulation provided by the present invention is inducing and/or enhancing immune response against malaria antigen in a mammal. Thus, in one embodiment, the present invention provides a method of inducing and/or enhancing immune response against malaria antigen in a mammal, comprising administering an effective amount of the composition as described above to the mammal.

[0134] Given the symptom of patients infected with Chikungunya or Venezuelan equine encephalitis together with unusual big molecule of Chikungunya or Venezuelan equine encephalitis, this VLP can act effectively and efficiently to target macrophage and its composition such as cytokines and immunomodulative compounds.

[0135] In one aspect, the present invention provides a method of presenting an antigen on macrophage, comprising administering the Chikungunya or Venezuelan equine encephalitis virus like particle as described above and/or the nucleic acid molecule as described above to a mammal. The Chikungunya or Venezuelan equine encephalitis virus like particle provided by the present invention is good to target macrophage. In one embodiment, the Chikungunya or Venezuelan equine encephalitis virus like particle provided by the present invention is a kind of delivery system of the at least one antigen, which is comprised in the Chikungunya or Venezuelan equine encephalitis virus like particle, to macrophage.

[0136] In one embodiment, the present invention provides a method for producing Chikungunya or Venezuelan equine encephalitis virus like particle provided in the first aspect of the present invention, comprising preparing a vector designed for expression of said particle; culturing a cell which is transfected with said vector to express said particle; and recovering said particle. In this embodiment, transfection can be conducted using a conventional method. Cells using for the transfection may be 293 cells. Recovering VLP may include collecting a conditioned medium after cells are transfected with a vector, and may further include purify VLP from the conditioned medium using ultracentrifugation.

[0137] The present invention will be described in detail with reference to the following example, which, however, is not intended to limit the scope of the present invention.

EXAMPLES

Example 1

Preparation of Chikungunya Virus (CHIKV) Like Particle Comprising a Virus Structural Polypeptide and a Fragment of Malaria Antigen

[0138] The following polynucleotides of malaria CSP1 proteins are used. N terminal linker is SGG and C terminal linker is GGS.

TABLE-US-00010 VLP74 (6 repeat of NPNA amino acid sequence) (SEQ ID No.: 46) Sggnpnanpnanpnanpnanpnanpnaggs (SEQ ID No.: 47) (Tccggaggaaacccgaatgccaatcccaacgcgaaccccaatgcta acccaaatgccaacccaaacgccaaccccaacgctggtggatcc) VLP78 (25 repeat of NPNA amino acid sequence) (SEQ ID No.: 48) Sggnpnanpnanpnanpnanpnanpnvdpnanpnanpnanpnanpna npnanpnanpnanpnanpnanpnanpnanpnanpnanpnanpnanpn anpnanpnaggs (SEQ ID No.: 49) (tccggaggaaacccgaatgccaatcccaacgcgaaccccaacgcta accccaacgccaatccgaatgcaaacccgaacgttgacccaaacgcc aacccgaatgccaatcccaacgcgaaccccaatgctaacccaaatgc caacccaaacgccaaccccaacgctaatccaaacgccaaccctaacg ccaatcccaacgcgaatcctaacgctaatcccaacgcaaatcccaat gctaatccgaacgcgaaccctaatgcaaaccccaacgccaacccgaa cgctaacccgaacgctaatcccaacgccggtggatcc)

[0139] The respective polynucleotides was inserted between the codons encoding Ser at 531-position and Asn at 532-position of SEQ ID Nos.15 or 16 (SEQ ID Nos.1 or 2) to construct a plasmid (hereinafter referred to as CHIKV-VLP74 or 78) for expressing Chikungunya virus like particle where the modified VLP74 or 78-derived peptide is inserted into E2 of Chikungunya virus structural polypeptide.

[0140] 293F cells (Lifetechnology) were transfected with the plasmid using PEI (GE Healthcare) or GeneX (ATCC). 4 days after the transfection, the conditioned medium was collected and centrifuged at 3000 rpm for 15 minutes to separate it from cells. The supernatant was filtrated using 0.45 .mu.m filter to obtain virus like particles. The virus like particles were concentrated using TFF column and purified using QXL column (GE Healthcare) to obtain purified virus like particles. When animals were immunized with virus like particles, the purified virus like particles were further concentrated using spin column (Molecular Weight-cutoff: 100 kDa) to prepare the virus like particles for the immunization.

[0141] The expression of VLP comprising VLP74 or 78 conjugated with Chikungunya virus structural polypeptide was confirmed by Western Blot using an antibody specific for CHIKV (ATCC: VR-1241AF) and an antibody specific for VLP74 or 78.

Example 2

Preparation of Venezuelan Equine Encephalitis Virus (VEEV) Like Particle Comprising a Virus Structural Polypeptide and a Fragment of Malaria Antigen

[0142] The same polynucleotides of malaria CSP1 proteins (VLP74 and VLP78) used in EXAMPLE 1 are used. N terminal linker is SGG and C terminal linker is GGS.

[0143] The respective polynucleotides was inserted between the codons encoding Ser at 518-position and Ser at 519-position of SEQ ID Nos.19 or 20 (SEQ ID No.3) to construct a plasmid (hereinafter referred to as VEEV-VLP74 or 78) for expressing Venezuelan equine encephalitis virus like particle where the modified VLP74 or 78-derived peptide is inserted into E2 of Venezuelan equine encephalitis virus structural polypeptide.

[0144] 293F cells (Lifetechnology) were transfected with the plasmid using PEI (GE Healthcare) or GeneX (ATCC). 4 days after the transfection, the conditioned medium was collected and centrifuged at 3000 rpm for 15 minutes to separate it from cells. The supernatant was filtrated using 0.45 .mu.m filter to obtain virus like particles. The virus like particles were concentrated using TFF column and purified using QXL column (GE Healthcare) to obtain purified virus like particles. When animals were immunized with virus like particles, the purified virus like particles were further concentrated using spin column (Molecular Weight-cutoff: 100 kDa) to prepare the virus like particles for the immunization.

[0145] The expression of VLP comprising VLP 74 or 78 conjugated with Venezuelan equine encephalitis virus structural polypeptide was confirmed by Western Blot using an antibody specific for VEEV and an antibody specific for VLP74 or 78.

Example 3

Immunogenicity in Non-Human Primate (Monkey)

[0146] The monkeys were immunized with .times.25-CHI (80 ug) at 0 week and .times.6-VEE (80 ug) at 3 week by intramuscular injection with or without adjuvant (Sigma Adjuvant System, Sigma, S6322). .times.25-CHI means 25 times malaria CSP repeat amino acid NPNA on CHIKV VLP particle (VLP78_15). .times.6-VEE means 6 times malaria CSP repeat amino acid NPNA on VEEV VLP particle (VLP74_25). The blood is taken at 2 and 5 weeks after the first immunization.

[0147] 96 well ELISA plate were coated with 50 ng of Recombinant Circumsporozoite Protein (rCSP) (Reagent Proteins, ATG-422) in 100 ul PBS buffer pre well. The Plates after 2 hours incubation were washed three times TBS buffer containing 0.05% Tween-20 and blocked with TBS buffer containing 0.05% Tween-20 and 5% dry milk. The heat inactivated diluted serum from monkeys were added in the blocking buffer and incubated for 1 h at room temperature. After washing three times, peroxidase labeled goat anti-human IgG or anti-mouse IgG was added at 1:4000 dilution and incubated for 1 h at room temperature. After washing three times, Peroxidase substrate was added for development and incubated for 10 mins and 2N H2SO4 was added to stop the development. The data were analyzed using Gen5 (BioTek) and GraphPad Prism6 (GraphPad software Inc).

[0148] The Immunogenicities are shown in FIGS. 4 to 6.

[0149] Induction of antibodies against CSP was found in the serum of all monkeys immunized with Malaria VLPs (see FIG. 4). The mean OD values indicating titer of antibodies against CSP is shown in FIG. 5. FIG. 5 shows that the serum from immunized monkeys induced high titer of antibodies against CSP.

[0150] As seen in FIG. 6, higher titer of antibodies against CSP was achieved when CHIKV VLP particle comprising NPNA and VEEV VLP particle comprising NPNA were used for the priming immunization and boosting immunization, respectively, compared to when only CHIKV VLP particle comprising NPNA was used for both of the priming immunization and the boosting immunization. In addition, FIG. 6 shows that use of adjuvant further enhanced the titer of antibodies against CSP. Further, FIG. 6 shows that administration of 25-repeats of NPNA induces higher titer of antibodies against CSP compared to administration of 6-repeats of NPNA.

[0151] The anti-Pf CSP antibody titer in the serum from the monkeys immunized with .times.25-CHI (80 ug) at 0 week and .times.6-VEE (80 ug) at 3 week without using adjuvant was measured by ELISA at Malaria Serology Lab Malaria Vaccine Branch, WRAIR. In the ELISA performed at Malaria Serology Lab Malaria Vaccine Branch, WRAIR, the plates were coated with CSPrp ((NPNA)6 Peptide) [0.2 .mu.g/.mu.L] (Supplier: Eurogentec EP070034) and Goat .alpha.-Human IgG (KPL/074-1002 LOT#120714) was used as 2nd antibody. The final titer was determined by the dilution factor that yields an OD of 1.0 (414 nm).

[0152] As a result, the anti-Pf CSP antibody titer in the serum from the monkeys was enhanced after 2nd immunization compared to 1st immunization (see Table 2). Compared to the anti-Pf CSP antibody titer in the serum from the monkeys immunized with RTS,S (GlaxoSmithKline), the anti-Pf CSP antibody titer in the serum from the monkeys immunized with .times.25-CHI (80 ug) and .times.6-VEE (80 ug) in the absence of adjuvant was considered to be higher even though RTS,S (GlaxoSmithKline) contains adjuvant.

TABLE-US-00011 TABLE 2 After 1st After 2nd immunization immunization Animal No. Week 2 Week 5 1 8990 29420 2 48210 44100 3 80400 51230 4 16260 19640 Geometric mean 27359 33801

Example 4

Immunogenicity in Mouse

[0153] The mice immunized with 10 ug of VLP78_15 at week 0, 10 ug of VLP74_25 at week 3 and 10 ug of VLP78_15 at week 6 with or without adjuvant (Sigma Adjuvant System, Sigma, S6322) by intramuscular injection.

[0154] The anti-Pf CSP antibody titer in the serum from the immunized mice were measured by ELISA at Malaria Serology Lab Malaria Vaccine Branch, WRAIR, where plates were coated with CSPrp ((NPNA)6 Peptide) [0.2 .mu.g/.mu.L] (Supplier: Eurogentec EP070034) and Goat .alpha.-Mouse IgG (KPL/074-1806 LOT#100737) is used as 2nd antibody to detect the antibodies in the serum.

[0155] The final titer was determined by the dilution factor that yields an OD of 1.0 (414 nm).

[0156] The Immunogenicity are shown in Tables 3 and 4.

[0157] Tables 3 and 4 show that higher titer of antibodies against CSP was achieved after immunizing virus like particle three times. In addition, Tables 3 and 4 show that use of adjuvant enhanced the titer of antibodies against CSP.

TABLE-US-00012 TABLE 3 Mouse Week 3 (after 1.sup.st immunization) Mouse No. VLP VLP + Adjuvant 1 6070 QNS 2 5850 13680 3 9610 7610 4 5440 23370 5 16320 27390 Geometric Mean 7875 16066 QNS = Quantity not sufficient to test

TABLE-US-00013 TABLE 4 Mouse Week 9 (after 1st immunization) Mouse No. VLP VLP + Adjuvant 1 35510 729000 2 15040 197800 3 41650 106700 4 37250 436000 5 48200 497600 Geometric Mean 33134 319666

Example 5

Immunogenicity of P. yoelii CSP Inserted VLP in Mice

[0158] QGPGAP seen in the rodent malaria CSP (P.yoelii CSP) was used as an antigen. 6.times. QGPGAP was inserted into CHIKV VLP. The mice were immunized with the CHIKV VLP 2 times at 0 and 8 week (20 ug VLP per mouse) by intramuscle injection with or without Adjuvant Ribi.

[0159] Immunogenicity was confirmed at 4, 6, 10 and 14 weeks after the first immunization. The anti-P. yoelii

[0160] CSP antibody were measured by ELISA. The ELISA was performed in the same way as ELISA described in Example 3 except that the plates were coated with P. Yoelii CSP repeat sequence peptide at 0.1 ng/.mu.l. The secondary antibody was anti-mouse IgG HRP (Cell signal, #7076S). The results are shown in FIGS. 7-9.

[0161] FIGS. 7-9 show that higher titer of antibodies against CSP was achieved by intramuscler administration of CHIKV VLP comprising 6.times. QGPGAP. In addition, FIGS. 7-9 show that use of adjuvant enhanced the titer of antibodies against CSP.

Example 6

Protection of Mice Against Malaria by Intramuscle Injection of CHIKV VLP Comprising P. yoelii CSP Epitope: 6.times. QGPGAP

[0162] 6.times. QGPGAP was inserted into CHIKV VLP. The mice (n=5) were immunized with the CHIKV VLP 2 times at 0 and 8 week (20 ug VLP per mouse) by intramuscle injection with or without Adjuvant Ribi (see FIG. 10). Rodent malaria: P. yoelii-challenge (i.v.) was conducted at 17 weeks (see FIG. 10).

[0163] Malaria infection was confirmed by PCR. Genomic DNA was purified from the mice blood day 6 after challenge. 18S malaria DNA was amplified by PCR. FIG. 11 shows results of the PCR, indicating that all of 5 control mice (PBS injection) were infected with malaria; all of 5 mice immunized with Control VLP were infected with malaria; among 5 mice immunized with CHIKV VLP comprising 6.times. QGPGAP, 2 mice were infected with malaria and 3 mice were not infected with malaria; and among 5 mice immunized with CHIKV VLP comprising 6.times. QGPGAP with adjuvant: Ribi, 1 mouse was infected with malaria and 4 mice were not infected with malaria.

Example 7

Preparation of Vaccine Composition Comprising Chikungunya Virus (CHIKV) Like Particle Comprising NPNA Repeat or Venezuelan Equine Encephalitis Virus (VEEV) Like Particle Comprising NPNA Repeat

[0164] Chikungunya virus (CHIKV) like particle comprising 6.times. or 25.times. NPNA was prepared according to Example 1, and Venezuelan equine encephalitis virus (VEEV) like particle comprising 6.times. NPNA was prepared according to Example 2. To prepare a vaccine composition, 80 .mu.g of each of the prepared particles was mixed with 1 ml of Sucrose Phosphate Solution, pH 7.2, Endotoxin Free (Teknova, SP buffer).

Sequence CWU 1

1

5611248PRTChikungunya virus 1Met Glu Phe Ile Pro Thr Gln Thr Phe Tyr Asn Arg Arg Tyr Gln Pro 1 5 10 15 Arg Pro Trp Thr Pro Arg Pro Thr Ile Gln Val Ile Arg Pro Arg Pro 20 25 30 Arg Pro Gln Arg Gln Ala Gly Gln Leu Ala Gln Leu Ile Ser Ala Val 35 40 45 Asn Lys Leu Thr Met Arg Ala Val Pro Gln Gln Lys Pro Arg Arg Asn 50 55 60 Arg Lys Asn Lys Lys Gln Lys Gln Lys Gln Gln Ala Pro Gln Asn Asn 65 70 75 80 Thr Asn Gln Lys Lys Gln Pro Pro Lys Lys Lys Pro Ala Gln Lys Lys 85 90 95 Lys Lys Pro Gly Arg Arg Glu Arg Met Cys Met Lys Ile Glu Asn Asp 100 105 110 Cys Ile Phe Glu Val Lys His Glu Gly Lys Val Thr Gly Tyr Ala Cys 115 120 125 Leu Val Gly Asp Lys Val Met Lys Pro Ala His Val Lys Gly Thr Ile 130 135 140 Asp Asn Ala Asp Leu Ala Lys Leu Ala Phe Lys Arg Ser Ser Lys Tyr 145 150 155 160 Asp Leu Glu Cys Ala Gln Ile Pro Val His Met Lys Ser Asp Ala Ser 165 170 175 Lys Phe Thr His Glu Lys Pro Glu Gly Tyr Tyr Asn Trp His His Gly 180 185 190 Ala Val Gln Tyr Ser Gly Gly Arg Phe Thr Ile Pro Thr Gly Ala Gly 195 200 205 Lys Pro Gly Asp Ser Gly Arg Pro Ile Phe Asp Asn Lys Gly Arg Val 210 215 220 Val Ala Ile Val Leu Gly Gly Ala Asn Glu Gly Ala Arg Thr Ala Leu 225 230 235 240 Ser Val Val Thr Trp Asn Lys Asp Ile Val Thr Lys Ile Thr Pro Glu 245 250 255 Gly Ala Glu Glu Trp Ser Leu Ala Ile Pro Val Met Cys Leu Leu Ala 260 265 270 Asn Thr Thr Phe Pro Cys Ser Gln Pro Pro Cys Thr Pro Cys Cys Tyr 275 280 285 Glu Lys Glu Pro Glu Glu Thr Leu Arg Met Leu Glu Asp Asn Val Met 290 295 300 Arg Pro Gly Tyr Tyr Gln Leu Leu Gln Ala Ser Leu Thr Cys Ser Pro 305 310 315 320 His Arg Gln Arg Arg Ser Thr Lys Asp Asn Phe Asn Val Tyr Lys Ala 325 330 335 Thr Arg Pro Tyr Leu Ala His Cys Pro Asp Cys Gly Glu Gly His Ser 340 345 350 Cys His Ser Pro Val Ala Leu Glu Arg Ile Arg Asn Glu Ala Thr Asp 355 360 365 Gly Thr Leu Lys Ile Gln Val Ser Leu Gln Ile Gly Ile Lys Thr Asp 370 375 380 Asp Ser His Asp Trp Thr Lys Leu Arg Tyr Met Asp Asn His Met Pro 385 390 395 400 Ala Asp Ala Glu Arg Ala Gly Leu Phe Val Arg Thr Ser Ala Pro Cys 405 410 415 Thr Ile Thr Gly Thr Met Gly His Phe Ile Leu Ala Arg Cys Pro Lys 420 425 430 Gly Glu Thr Leu Thr Val Gly Phe Thr Asp Ser Arg Lys Ile Ser His 435 440 445 Ser Cys Thr His Pro Phe His His Asp Pro Pro Val Ile Gly Arg Glu 450 455 460 Lys Phe His Ser Arg Pro Gln His Gly Lys Glu Leu Pro Cys Ser Thr 465 470 475 480 Tyr Val Gln Ser Thr Ala Ala Thr Thr Glu Glu Ile Glu Val His Met 485 490 495 Pro Pro Asp Thr Pro Asp Arg Thr Leu Met Ser Gln Gln Ser Gly Asn 500 505 510 Val Lys Ile Thr Val Asn Gly Gln Thr Val Arg Tyr Lys Cys Asn Cys 515 520 525 Gly Gly Ser Asn Glu Gly Leu Thr Thr Thr Asp Lys Val Ile Asn Asn 530 535 540 Cys Lys Val Asp Gln Cys His Ala Ala Val Thr Asn His Lys Lys Trp 545 550 555 560 Gln Tyr Asn Ser Pro Leu Val Pro Arg Asn Ala Glu Leu Gly Asp Arg 565 570 575 Lys Gly Lys Ile His Ile Pro Phe Pro Leu Ala Asn Val Thr Cys Arg 580 585 590 Val Pro Lys Ala Arg Asn Pro Thr Val Thr Tyr Gly Lys Asn Gln Val 595 600 605 Ile Met Leu Leu Tyr Pro Asp His Pro Thr Leu Leu Ser Tyr Arg Asn 610 615 620 Met Gly Glu Glu Pro Asn Tyr Gln Glu Glu Trp Val Met His Lys Lys 625 630 635 640 Glu Val Val Leu Thr Val Pro Thr Glu Gly Leu Glu Val Thr Trp Gly 645 650 655 Asn Asn Glu Pro Tyr Lys Tyr Trp Pro Gln Leu Ser Thr Asn Gly Thr 660 665 670 Ala His Gly His Pro His Glu Ile Ile Leu Tyr Tyr Tyr Glu Leu Tyr 675 680 685 Pro Thr Met Thr Val Val Val Val Ser Val Ala Thr Phe Ile Leu Leu 690 695 700 Ser Met Val Gly Met Ala Ala Gly Met Cys Met Cys Ala Arg Arg Arg 705 710 715 720 Cys Ile Thr Pro Tyr Glu Leu Thr Pro Gly Ala Thr Val Pro Phe Leu 725 730 735 Leu Ser Leu Ile Cys Cys Ile Arg Thr Ala Lys Ala Ala Thr Tyr Gln 740 745 750 Glu Ala Ala Ile Tyr Leu Trp Asn Glu Gln Gln Pro Leu Phe Trp Leu 755 760 765 Gln Ala Leu Ile Pro Leu Ala Ala Leu Ile Val Leu Cys Asn Cys Leu 770 775 780 Arg Leu Leu Pro Cys Cys Cys Lys Thr Leu Ala Phe Leu Ala Val Met 785 790 795 800 Ser Val Gly Ala His Thr Val Ser Ala Tyr Glu His Val Thr Val Ile 805 810 815 Pro Asn Thr Val Gly Val Pro Tyr Lys Thr Leu Val Asn Arg Pro Gly 820 825 830 Tyr Ser Pro Met Val Leu Glu Met Glu Leu Leu Ser Val Thr Leu Glu 835 840 845 Pro Thr Leu Ser Leu Asp Tyr Ile Thr Cys Glu Tyr Lys Thr Val Ile 850 855 860 Pro Ser Pro Tyr Val Lys Cys Cys Gly Thr Ala Glu Cys Lys Asp Lys 865 870 875 880 Asn Leu Pro Asp Tyr Ser Cys Lys Val Phe Thr Gly Val Tyr Pro Phe 885 890 895 Met Trp Gly Gly Ala Tyr Cys Phe Cys Asp Ala Glu Asn Thr Gln Leu 900 905 910 Ser Glu Ala His Val Glu Lys Ser Glu Ser Cys Lys Thr Glu Phe Ala 915 920 925 Ser Ala Tyr Arg Ala His Thr Ala Ser Ala Ser Ala Lys Leu Arg Val 930 935 940 Leu Tyr Gln Gly Asn Asn Ile Thr Val Thr Ala Tyr Ala Asn Gly Asp 945 950 955 960 His Ala Val Thr Val Lys Asp Ala Lys Phe Ile Val Gly Pro Met Ser 965 970 975 Ser Ala Trp Thr Pro Phe Asp Asn Lys Ile Val Val Tyr Lys Gly Asp 980 985 990 Val Tyr Asn Met Asp Tyr Pro Pro Phe Gly Ala Gly Arg Pro Gly Gln 995 1000 1005 Phe Gly Asp Ile Gln Ser Arg Thr Pro Glu Ser Lys Asp Val Tyr 1010 1015 1020 Ala Asn Thr Gln Leu Val Leu Gln Arg Pro Ala Val Gly Thr Val 1025 1030 1035 His Val Pro Tyr Ser Gln Ala Pro Ser Gly Phe Lys Tyr Trp Leu 1040 1045 1050 Lys Glu Arg Gly Ala Ser Leu Gln His Thr Ala Pro Phe Gly Cys 1055 1060 1065 Gln Ile Ala Thr Asn Pro Val Arg Ala Val Asn Cys Ala Val Gly 1070 1075 1080 Asn Met Pro Ile Ser Ile Asp Ile Pro Glu Ala Ala Phe Thr Arg 1085 1090 1095 Val Val Asp Ala Pro Ser Leu Thr Asp Met Ser Cys Glu Val Pro 1100 1105 1110 Ala Cys Thr His Ser Ser Asp Phe Gly Gly Val Ala Ile Ile Lys 1115 1120 1125 Tyr Ala Ala Ser Lys Lys Gly Lys Cys Ala Val His Ser Met Thr 1130 1135 1140 Asn Ala Val Thr Ile Arg Glu Ala Glu Ile Glu Val Glu Gly Asn 1145 1150 1155 Ser Gln Leu Gln Ile Ser Phe Ser Thr Ala Leu Ala Ser Ala Glu 1160 1165 1170 Phe Arg Val Gln Val Cys Ser Thr Gln Val His Cys Ala Ala Glu 1175 1180 1185 Cys His Pro Pro Lys Asp His Ile Val Asn Tyr Pro Ala Ser His 1190 1195 1200 Thr Thr Leu Gly Val Gln Asp Ile Ser Ala Thr Ala Met Ser Trp 1205 1210 1215 Val Gln Lys Ile Thr Gly Gly Val Gly Leu Val Val Ala Val Ala 1220 1225 1230 Ala Leu Ile Leu Ile Val Val Leu Cys Val Ser Phe Ser Arg His 1235 1240 1245 21248PRTChikungunya virus 2Met Glu Phe Ile Pro Thr Gln Thr Phe Tyr Asn Arg Arg Tyr Gln Pro 1 5 10 15 Arg Pro Trp Ala Pro Arg Pro Thr Ile Gln Val Ile Arg Pro Arg Pro 20 25 30 Arg Pro Gln Arg Gln Ala Gly Gln Leu Ala Gln Leu Ile Ser Ala Val 35 40 45 Asn Lys Leu Thr Met Arg Ala Val Pro Gln Gln Lys Pro Arg Arg Asn 50 55 60 Arg Lys Asn Lys Lys Gln Arg Gln Lys Lys Gln Ala Pro Gln Asn Asp 65 70 75 80 Pro Lys Gln Lys Lys Gln Pro Pro Gln Lys Lys Pro Ala Gln Lys Lys 85 90 95 Lys Lys Pro Gly Arg Arg Glu Arg Met Cys Met Lys Ile Glu Asn Asp 100 105 110 Cys Ile Phe Glu Val Lys His Glu Gly Lys Val Met Gly Tyr Ala Cys 115 120 125 Leu Val Gly Asp Lys Val Met Lys Pro Ala His Val Lys Gly Thr Ile 130 135 140 Asp Asn Ala Asp Leu Ala Lys Leu Ala Phe Lys Arg Ser Ser Lys Tyr 145 150 155 160 Asp Leu Glu Cys Ala Gln Ile Pro Val His Met Lys Ser Asp Ala Ser 165 170 175 Lys Phe Thr His Glu Lys Pro Glu Gly Tyr Tyr Asn Trp His His Gly 180 185 190 Ala Val Gln Tyr Ser Gly Gly Arg Phe Thr Ile Pro Thr Gly Ala Gly 195 200 205 Lys Pro Gly Asp Ser Gly Arg Pro Ile Phe Asp Asn Lys Gly Arg Val 210 215 220 Val Ala Ile Val Leu Gly Gly Ala Asn Glu Gly Ala Arg Thr Ala Leu 225 230 235 240 Ser Val Val Thr Trp Asn Lys Asp Ile Val Thr Lys Ile Thr Pro Glu 245 250 255 Gly Ala Glu Glu Trp Ser Leu Ala Leu Pro Val Leu Cys Leu Leu Ala 260 265 270 Asn Thr Thr Phe Pro Cys Ser Gln Pro Pro Cys Thr Pro Cys Cys Tyr 275 280 285 Glu Lys Glu Pro Glu Ser Thr Leu Arg Met Leu Glu Asp Asn Val Met 290 295 300 Arg Pro Gly Tyr Tyr Gln Leu Leu Lys Ala Ser Leu Thr Cys Ser Pro 305 310 315 320 His Arg Gln Arg Arg Ser Thr Lys Asp Asn Phe Asn Val Tyr Lys Ala 325 330 335 Thr Arg Pro Tyr Leu Ala His Cys Pro Asp Cys Gly Glu Gly His Ser 340 345 350 Cys His Ser Pro Ile Ala Leu Glu Arg Ile Arg Asn Glu Ala Thr Asp 355 360 365 Gly Thr Leu Lys Ile Gln Val Ser Leu Gln Ile Gly Ile Lys Thr Asp 370 375 380 Asp Ser His Asp Trp Thr Lys Leu Arg Tyr Met Asp Ser His Thr Pro 385 390 395 400 Ala Asp Ala Glu Arg Ala Gly Leu Leu Val Arg Thr Ser Ala Pro Cys 405 410 415 Thr Ile Thr Gly Thr Met Gly His Phe Ile Leu Ala Arg Cys Pro Lys 420 425 430 Gly Glu Thr Leu Thr Val Gly Phe Thr Asp Ser Arg Lys Ile Ser His 435 440 445 Thr Cys Thr His Pro Phe His His Glu Pro Pro Val Ile Gly Arg Glu 450 455 460 Arg Phe His Ser Arg Pro Gln His Gly Lys Glu Leu Pro Cys Ser Thr 465 470 475 480 Tyr Val Gln Ser Thr Ala Ala Thr Ala Glu Glu Ile Glu Val His Met 485 490 495 Pro Pro Asp Thr Pro Asp Arg Thr Leu Met Thr Gln Gln Ser Gly Asn 500 505 510 Val Lys Ile Thr Val Asn Gly Gln Thr Val Arg Tyr Lys Cys Asn Cys 515 520 525 Gly Gly Ser Asn Glu Gly Leu Thr Thr Thr Asp Lys Val Ile Asn Asn 530 535 540 Cys Lys Ile Asp Gln Cys His Ala Ala Val Thr Asn His Lys Asn Trp 545 550 555 560 Gln Tyr Asn Ser Pro Leu Val Pro Arg Asn Ala Glu Leu Gly Asp Arg 565 570 575 Lys Gly Lys Ile His Ile Pro Phe Pro Leu Ala Asn Val Thr Cys Arg 580 585 590 Val Pro Lys Ala Arg Asn Pro Thr Val Thr Tyr Gly Lys Asn Gln Val 595 600 605 Thr Met Leu Leu Tyr Pro Asp His Pro Thr Leu Leu Ser Tyr Arg Asn 610 615 620 Met Gly Gln Glu Pro Asn Tyr His Glu Glu Trp Val Thr His Lys Lys 625 630 635 640 Glu Val Thr Leu Thr Val Pro Thr Glu Gly Leu Glu Val Thr Trp Gly 645 650 655 Asn Asn Glu Pro Tyr Lys Tyr Trp Pro Gln Met Ser Thr Asn Gly Thr 660 665 670 Ala His Gly His Pro His Glu Ile Ile Leu Tyr Tyr Tyr Glu Leu Tyr 675 680 685 Pro Thr Met Thr Val Val Ile Val Ser Val Ala Ser Phe Val Leu Leu 690 695 700 Ser Met Val Gly Thr Ala Val Gly Met Cys Val Cys Ala Arg Arg Arg 705 710 715 720 Cys Ile Thr Pro Tyr Glu Leu Thr Pro Gly Ala Thr Val Pro Phe Leu 725 730 735 Leu Ser Leu Leu Cys Cys Val Arg Thr Thr Lys Ala Ala Thr Tyr Tyr 740 745 750 Glu Ala Ala Ala Tyr Leu Trp Asn Glu Gln Gln Pro Leu Phe Trp Leu 755 760 765 Gln Ala Leu Ile Pro Leu Ala Ala Leu Ile Val Leu Cys Asn Cys Leu 770 775 780 Lys Leu Leu Pro Cys Cys Cys Lys Thr Leu Ala Phe Leu Ala Val Met 785 790 795 800 Ser Ile Gly Ala His Thr Val Ser Ala Tyr Glu His Val Thr Val Ile 805 810 815 Pro Asn Thr Val Gly Val Pro Tyr Lys Thr Leu Val Asn Arg Pro Gly 820 825 830 Tyr Ser Pro Met Val Leu Glu Met Glu Leu Gln Ser Val Thr Leu Glu 835 840 845 Pro Thr Leu Ser Leu Asp Tyr Ile Thr Cys Glu Tyr Lys Thr Val Ile 850 855 860 Pro Ser Pro Tyr Val Lys Cys Cys Gly Thr Ala Glu Cys Lys Asp Lys 865 870 875 880 Ser Leu Pro Asp Tyr Ser Cys Lys Val Phe Thr Gly Val Tyr Pro Phe 885 890 895 Met Trp Gly Gly Ala Tyr Cys Phe Cys Asp Ala Glu Asn Thr Gln Leu 900 905 910 Ser Glu Ala His Val Glu Lys Ser Glu Ser Cys Lys Thr Glu Phe Ala 915 920 925 Ser Ala Tyr Arg Ala His Thr Ala Ser Ala Ser Ala Lys Leu Arg Val 930 935 940 Leu Tyr Gln Gly Asn Asn Ile Thr Val Ala Ala Tyr Ala Asn Gly Asp 945 950 955 960 His Ala Val Thr Val Lys Asp Ala Lys Phe Val Val Gly Pro Met Ser 965 970 975 Ser Ala Trp Thr Pro Phe Asp Asn Lys Ile Val Val Tyr Lys Gly Asp 980 985 990 Val Tyr Asn Met Asp Tyr Pro Pro Phe Gly Ala Gly Arg Pro Gly Gln 995 1000 1005 Phe Gly Asp Ile Gln Ser Arg Thr Pro Glu Ser Lys Asp Val Tyr 1010 1015 1020 Ala Asn Thr Gln Leu Val Leu Gln Arg Pro Ala Ala Gly Thr Val 1025 1030 1035

His Val Pro Tyr Ser Gln Ala Pro Ser Gly Phe Lys Tyr Trp Leu 1040 1045 1050 Lys Glu Arg Gly Ala Ser Leu Gln His Thr Ala Pro Phe Gly Cys 1055 1060 1065 Gln Ile Ala Thr Asn Pro Val Arg Ala Val Asn Cys Ala Val Gly 1070 1075 1080 Asn Ile Pro Ile Ser Ile Asp Ile Pro Asp Ala Ala Phe Thr Arg 1085 1090 1095 Val Val Asp Ala Pro Ser Val Thr Asp Met Ser Cys Glu Val Pro 1100 1105 1110 Ala Cys Thr His Ser Ser Asp Phe Gly Gly Val Ala Ile Ile Lys 1115 1120 1125 Tyr Thr Ala Ser Lys Lys Gly Lys Cys Ala Val His Ser Met Thr 1130 1135 1140 Asn Ala Val Thr Ile Arg Glu Ala Asp Val Glu Val Glu Gly Asn 1145 1150 1155 Ser Gln Leu Gln Ile Ser Phe Ser Thr Ala Leu Ala Ser Ala Glu 1160 1165 1170 Phe Arg Val Gln Val Cys Ser Thr Gln Val His Cys Ala Ala Ala 1175 1180 1185 Cys His Pro Pro Lys Asp His Ile Val Asn Tyr Pro Ala Ser His 1190 1195 1200 Thr Thr Leu Gly Val Gln Asp Ile Ser Thr Thr Ala Met Ser Trp 1205 1210 1215 Val Gln Lys Ile Thr Gly Gly Val Gly Leu Ile Val Ala Val Ala 1220 1225 1230 Ala Leu Ile Leu Ile Val Val Leu Cys Val Ser Phe Ser Arg His 1235 1240 1245 31255PRTVenesuelan equine encephalitis virus 3Met Phe Pro Phe Gln Pro Met Tyr Pro Met Gln Pro Met Pro Tyr Arg 1 5 10 15 Asn Pro Phe Ala Ala Pro Arg Arg Pro Trp Phe Pro Arg Thr Asp Pro 20 25 30 Phe Leu Ala Met Gln Val Gln Glu Leu Thr Arg Ser Met Ala Asn Leu 35 40 45 Thr Phe Lys Gln Arg Arg Asp Ala Pro Pro Glu Gly Pro Ser Ala Ala 50 55 60 Lys Pro Lys Lys Glu Ala Ser Gln Lys Gln Lys Gly Gly Gly Gln Gly 65 70 75 80 Lys Lys Lys Lys Asn Gln Gly Lys Lys Lys Ala Lys Thr Gly Pro Pro 85 90 95 Asn Pro Lys Ala Gln Asn Gly Asn Lys Lys Lys Thr Asn Lys Lys Pro 100 105 110 Gly Lys Arg Gln Arg Met Val Met Lys Leu Glu Ser Asp Lys Thr Phe 115 120 125 Pro Ile Met Leu Glu Gly Lys Ile Asn Gly Tyr Ala Cys Val Val Gly 130 135 140 Gly Lys Leu Phe Arg Pro Met His Val Glu Gly Lys Ile Asp Asn Asp 145 150 155 160 Val Leu Ala Ala Leu Lys Thr Lys Lys Ala Ser Lys Tyr Asp Leu Glu 165 170 175 Tyr Ala Asp Val Pro Gln Asn Met Arg Ala Asp Thr Phe Lys Tyr Thr 180 185 190 His Glu Lys Pro Gln Gly Tyr Tyr Ser Trp His His Gly Ala Val Gln 195 200 205 Tyr Glu Asn Gly Arg Phe Thr Val Pro Lys Gly Val Gly Ala Lys Gly 210 215 220 Asp Ser Gly Arg Pro Ile Leu Asp Asn Gln Gly Arg Val Val Ala Ile 225 230 235 240 Val Leu Gly Gly Val Asn Glu Gly Ser Arg Thr Ala Leu Ser Val Val 245 250 255 Met Trp Asn Glu Lys Gly Val Thr Val Lys Tyr Thr Pro Glu Asn Cys 260 265 270 Glu Gln Trp Ser Leu Val Thr Thr Met Cys Leu Leu Ala Asn Val Thr 275 280 285 Phe Pro Cys Ala Gln Pro Pro Ile Cys Tyr Asp Arg Lys Pro Ala Glu 290 295 300 Thr Leu Ala Met Leu Ser Val Asn Val Asp Asn Pro Gly Tyr Asp Glu 305 310 315 320 Leu Leu Glu Ala Ala Val Lys Cys Pro Gly Arg Lys Arg Arg Ser Thr 325 330 335 Glu Glu Leu Phe Asn Glu Tyr Lys Leu Thr Arg Pro Tyr Met Ala Arg 340 345 350 Cys Ile Arg Cys Ala Val Gly Ser Cys His Ser Pro Ile Ala Ile Glu 355 360 365 Ala Val Lys Ser Asp Gly His Asp Gly Tyr Val Arg Leu Gln Thr Ser 370 375 380 Ser Gln Tyr Gly Leu Asp Ser Ser Gly Asn Leu Lys Gly Arg Thr Met 385 390 395 400 Arg Tyr Asp Met His Gly Thr Ile Lys Glu Ile Pro Leu His Gln Val 405 410 415 Ser Leu Tyr Thr Ser Arg Pro Cys His Ile Val Asp Gly His Gly Tyr 420 425 430 Phe Leu Leu Ala Arg Cys Pro Ala Gly Asp Ser Ile Thr Met Glu Phe 435 440 445 Lys Lys Asp Ser Val Arg His Ser Cys Ser Val Pro Tyr Glu Val Lys 450 455 460 Phe Asn Pro Val Gly Arg Glu Leu Tyr Thr His Pro Pro Glu His Gly 465 470 475 480 Val Glu Gln Ala Cys Gln Val Tyr Ala His Asp Ala Gln Asn Arg Gly 485 490 495 Ala Tyr Val Glu Met His Leu Pro Gly Ser Glu Val Asp Ser Ser Leu 500 505 510 Val Ser Leu Ser Gly Ser Ser Val Thr Val Thr Pro Pro Asp Gly Thr 515 520 525 Ser Ala Leu Val Glu Cys Glu Cys Gly Gly Thr Lys Ile Ser Glu Thr 530 535 540 Ile Asn Lys Thr Lys Gln Phe Ser Gln Cys Thr Lys Lys Glu Gln Cys 545 550 555 560 Arg Ala Tyr Arg Leu Gln Asn Asp Lys Trp Val Tyr Asn Ser Asp Lys 565 570 575 Leu Pro Lys Ala Ala Gly Ala Thr Leu Lys Gly Lys Leu His Val Pro 580 585 590 Phe Leu Leu Ala Asp Gly Lys Cys Thr Val Pro Leu Ala Pro Glu Pro 595 600 605 Met Ile Thr Phe Gly Phe Arg Ser Val Ser Leu Lys Leu His Pro Lys 610 615 620 Asn Pro Thr Tyr Leu Ile Thr Arg Gln Leu Ala Asp Glu Pro His Tyr 625 630 635 640 Thr His Glu Leu Ile Ser Glu Pro Ala Val Arg Asn Phe Thr Val Thr 645 650 655 Glu Lys Gly Trp Glu Phe Val Trp Gly Asn His Pro Pro Lys Arg Phe 660 665 670 Trp Ala Gln Glu Thr Ala Pro Gly Asn Pro His Gly Leu Pro His Glu 675 680 685 Val Ile Thr His Tyr Tyr His Arg Tyr Pro Met Ser Thr Ile Leu Gly 690 695 700 Leu Ser Ile Cys Ala Ala Ile Ala Thr Val Ser Val Ala Ala Ser Thr 705 710 715 720 Trp Leu Phe Cys Arg Ser Arg Val Ala Cys Leu Thr Pro Tyr Arg Leu 725 730 735 Thr Pro Asn Ala Arg Ile Pro Phe Cys Leu Ala Val Leu Cys Cys Ala 740 745 750 Arg Thr Ala Arg Ala Glu Thr Thr Trp Glu Ser Leu Asp His Leu Trp 755 760 765 Asn Asn Asn Gln Gln Met Phe Trp Ile Gln Leu Leu Ile Pro Leu Ala 770 775 780 Ala Leu Ile Val Val Thr Arg Leu Leu Arg Cys Val Cys Cys Val Val 785 790 795 800 Pro Phe Leu Val Met Ala Gly Ala Ala Gly Ala Gly Ala Tyr Glu His 805 810 815 Ala Thr Thr Met Pro Ser Gln Ala Gly Ile Ser Tyr Asn Thr Ile Val 820 825 830 Asn Arg Ala Gly Tyr Ala Pro Leu Pro Ile Ser Ile Thr Pro Thr Lys 835 840 845 Ile Lys Leu Ile Pro Thr Val Asn Leu Glu Tyr Val Thr Cys His Tyr 850 855 860 Lys Thr Gly Met Asp Ser Pro Ala Ile Lys Cys Cys Gly Ser Gln Glu 865 870 875 880 Cys Thr Pro Thr Tyr Arg Pro Asp Glu Gln Cys Lys Val Phe Thr Gly 885 890 895 Val Tyr Pro Phe Met Trp Gly Gly Ala Tyr Cys Phe Cys Asp Thr Glu 900 905 910 Asn Thr Gln Val Ser Lys Ala Tyr Val Met Lys Ser Asp Asp Cys Leu 915 920 925 Ala Asp His Ala Glu Ala Tyr Lys Ala His Thr Ala Ser Val Gln Ala 930 935 940 Phe Leu Asn Ile Thr Val Gly Glu His Ser Ile Val Thr Thr Val Tyr 945 950 955 960 Val Asn Gly Glu Thr Pro Val Asn Phe Asn Gly Val Lys Ile Thr Ala 965 970 975 Gly Pro Leu Ser Thr Ala Trp Thr Pro Phe Asp Arg Lys Ile Val Gln 980 985 990 Tyr Ala Gly Glu Ile Tyr Asn Tyr Asp Phe Pro Glu Tyr Gly Ala Gly 995 1000 1005 Gln Pro Gly Ala Phe Gly Asp Ile Gln Ser Arg Thr Val Ser Ser 1010 1015 1020 Ser Asp Leu Tyr Ala Asn Thr Asn Leu Val Leu Gln Arg Pro Lys 1025 1030 1035 Ala Gly Ala Ile His Val Pro Tyr Thr Gln Ala Pro Ser Gly Phe 1040 1045 1050 Glu Gln Trp Lys Lys Asp Lys Ala Pro Ser Leu Lys Phe Thr Ala 1055 1060 1065 Pro Phe Gly Cys Glu Ile Tyr Thr Asn Pro Ile Arg Ala Glu Asn 1070 1075 1080 Cys Ala Val Gly Ser Ile Pro Leu Ala Phe Asp Ile Pro Asp Ala 1085 1090 1095 Leu Phe Thr Arg Val Ser Glu Thr Pro Thr Leu Ser Ala Ala Glu 1100 1105 1110 Cys Thr Leu Asn Glu Cys Val Tyr Ser Ser Asp Phe Gly Gly Ile 1115 1120 1125 Ala Thr Val Lys Tyr Ser Ala Ser Lys Ser Gly Lys Cys Ala Val 1130 1135 1140 His Val Pro Ser Gly Thr Ala Thr Leu Lys Glu Ala Ala Val Glu 1145 1150 1155 Leu Thr Glu Gln Gly Ser Ala Thr Ile His Phe Ser Thr Ala Asn 1160 1165 1170 Ile His Pro Glu Phe Arg Leu Gln Ile Cys Thr Ser Tyr Val Thr 1175 1180 1185 Cys Lys Gly Asp Cys His Pro Pro Lys Asp His Ile Val Thr His 1190 1195 1200 Pro Gln Tyr His Ala Gln Thr Phe Thr Ala Ala Val Ser Lys Thr 1205 1210 1215 Ala Trp Thr Trp Leu Thr Ser Leu Leu Gly Gly Ser Ala Val Ile 1220 1225 1230 Ile Ile Ile Gly Leu Val Leu Ala Thr Ile Val Ala Met Tyr Val 1235 1240 1245 Leu Thr Asn Gln Lys His Asn 1250 1255 42964DNAChikungunya virus 4atgagcctcg ccctcccggt cttgtgcctg ttggcaaaca ctacattccc ctgctctcag 60ccgccttgca caccctgctg ctacgaaaag gaaccggaaa gcaccttgcg catgcttgag 120gacaacgtga tgagacccgg atactaccag ctactaaaag catcgctgac ttgctctccc 180caccgccaaa gacgcagtac taaggacaat tttaatgtct ataaagccac aagaccatat 240ctagctcatt gtcctgactg cggagaaggg cattcgtgcc acagccctat cgcattggag 300cgcatcagaa atgaagcaac ggacggaacg ctgaaaatcc aggtctcttt gcagatcggg 360ataaagacag atgacagcca cgattggacc aagctgcgct atatggatag ccatacgcca 420gcggacgcgg agcgagccgg attgcttgta aggacttcag caccgtgcac gatcaccggg 480accatgggac actttattct cgcccgatgc ccgaaaggag agacgctgac agtgggattt 540acggacagca gaaagatcag ccacacatgc acacacccgt tccatcatga accacctgtg 600ataggtaggg agaggttcca ctctcgacca caacatggta aagagttacc ttgcagcacg 660tacgtgcaga gcaccgctgc cactgctgag gagatagagg tgcatatgcc cccagatact 720cctgaccgca cgctgatgac gcagcagtct ggcaacgtga agatcacagt taatgggcag 780acggtgcggt acaagtgcaa ctgcggtggc tcaaacgagg gactgacaac cacagacaaa 840gtgatcaata actgcaaaat tgatcagtgc catgctgcag tcactaatca caagaattgg 900caatacaact cccctttagt cccgcgcaac gctgaactcg gggaccgtaa aggaaagatc 960cacatcccat tcccattggc aaacgtgact tgcagagtgc caaaagcaag aaaccctaca 1020gtaacttacg gaaaaaacca agtcaccatg ctgctgtatc ctgaccatcc gacactcttg 1080tcttaccgta acatgggaca ggaaccaaat taccacgagg agtgggtgac acacaagaag 1140gaggttacct tgaccgtgcc tactgagggt ctggaggtca cttggggcaa caacgaacca 1200tacaagtact ggccgcagat gtctacgaac ggtactgctc atggtcaccc acatgagata 1260atcttgtact attatgagct gtaccccact atgactgtag tcattgtgtc ggtggcctcg 1320ttcgtgcttc tgtcgatggt gggcacagca gtgggaatgt gtgtgtgcgc acggcgcaga 1380tgcattacac catatgaatt aacaccagga gccactgttc ccttcctgct cagcctgcta 1440tgctgcgtca gaacgaccaa ggcggccaca tattacgagg ctgcggcata tctatggaac 1500gaacagcagc ccctgttctg gttgcaggct cttatcccgc tggccgcctt gatcgtcctg 1560tgcaactgtc tgaaactctt gccatgctgc tgtaagaccc tggctttttt agccgtaatg 1620agcatcggtg cccacactgt gagcgcgtac gaacacgtaa cagtgatccc gaacacggtg 1680ggagtaccgt ataagactct tgtcaacaga ccgggttaca gccccatggt gttggagatg 1740gagctacaat cagtcacctt ggaaccaaca ctgtcacttg actacatcac gtgcgagtac 1800aaaactgtca tcccctcccc gtacgtgaag tgctgtggta cagcagagtg caaggacaag 1860agcctaccag actacagctg caaggtcttt actggagtct acccatttat gtggggcggc 1920gcctactgct tttgcgacgc cgaaaatacg caattgagcg aggcacatgt agagaaatct 1980gaatcttgca aaacagagtt tgcatcggcc tacagagccc acaccgcatc ggcgtcggcg 2040aagctccgcg tcctttacca aggaaacaac attaccgtag ctgcctacgc taacggtgac 2100catgccgtca cagtaaagga cgccaagttt gtcgtgggcc caatgtcctc cgcctggaca 2160ccttttgaca acaaaatcgt ggtgtacaaa ggcgacgtct acaacatgga ctacccacct 2220tttggcgcag gaagaccagg acaatttggt gacattcaaa gtcgtacacc ggaaagtaaa 2280gacgtttatg ccaacactca gttggtacta cagaggccag cagcaggcac ggtacatgta 2340ccatactctc aggcaccatc tggcttcaag tattggctga aggaacgagg agcatcgcta 2400cagcacacgg caccgttcgg ttgccagatt gcgacaaacc cggtaagagc tgtaaattgc 2460gctgtgggga acataccaat ttccatcgac ataccggatg cggcctttac tagggttgtc 2520gatgcaccct ctgtaacgga catgtcatgc gaagtaccag cctgcactca ctcctccgac 2580tttgggggcg tcgccatcat caaatacaca gctagcaaga aaggtaaatg tgcagtacat 2640tcgatgacca acgccgttac cattcgagaa gccgacgtag aagtagaggg gaactcccag 2700ctgcaaatat ccttctcaac agccctggca agcgccgagt ttcgcgtgca agtgtgctcc 2760acacaagtac actgcgcagc cgcatgccac cctccaaagg accacatagt caattaccca 2820gcatcacaca ccacccttgg ggtccaggat atatccacaa cggcaatgtc ttgggtgcag 2880aagattacgg gaggagtagg attaattgtt gctgttgctg ccttaatttt aattgtggtg 2940ctatgcgtgt cgtttagcag gcac 296452964DNAChikungunya virus 5atgagtcttg ccatcccagt tatgtgcctg ttggcaaaca ccacgttccc ctgctcccag 60cccccttgca cgccctgctg ctacgaaaag gaaccggagg aaaccctacg catgcttgag 120gacaacgtca tgagacctgg gtactatcag ctgctacaag catccttaac atgttctccc 180caccgccagc gacgcagcac caaggacaac ttcaatgtct ataaagccac aagaccatac 240ttagctcact gtcccgactg tggagaaggg cactcgtgcc atagtcccgt agcactagaa 300cgcatcagaa atgaagcgac agacgggacg ctgaaaatcc aggtctcctt gcaaatcgga 360ataaagacgg atgacagcca cgattggacc aagctgcgtt atatggacaa ccacatgcca 420gcagacgcag agagggcggg gctatttgta agaacatcag caccgtgtac gattactgga 480acaatgggac acttcatcct ggcccgatgt ccaaaagggg aaactctgac ggtgggattc 540actgacagta ggaagattag tcactcatgt acgcacccat ttcaccacga ccctcctgtg 600ataggtcggg aaaaattcca ttcccgaccg cagcacggta aagagctacc ttgcagcacg 660tacgtgcaga gcaccgccgc aactaccgag gagatagagg tacacatgcc cccagacacc 720cctgatcgca cattaatgtc acaacagtcc ggcaacgtaa agatcacagt caatggccag 780acggtgcggt acaagtgtaa ttgcggtggc tcaaatgaag gactaacaac tacagacaaa 840gtgattaata actgcaaggt tgatcaatgt catgccgcgg tcaccaatca caaaaagtgg 900cagtataact cccctctggt cccgcgtaat gctgaacttg gggaccgaaa aggaaaaatt 960cacatcccgt ttccgctggc aaatgtaaca tgcagggtgc ctaaagcaag gaaccccacc 1020gtgacgtacg ggaaaaacca agtcatcatg ctactgtatc ctgaccaccc aacactcctg 1080tcctaccgga atatgggaga agaaccaaac tatcaagaag agtgggtgat gcataagaag 1140gaagtcgtgc taaccgtgcc gactgaaggg ctcgaggtca cgtggggcaa caacgagccg 1200tataagtatt ggccgcagtt atctacaaac ggtacagccc atggccaccc gcatgagata 1260attctgtatt attatgagct gtaccccact atgactgtag tagttgtgtc agtggccacg 1320ttcatactcc tgtcgatggt gggtatggca gcggggatgt gcatgtgtgc acgacgcaga 1380tgcatcacac cgtatgaact gacaccagga gctaccgtcc ctttcctgct tagcctaata 1440tgctgcatca gaacagctaa agcggccaca taccaagagg ctgcgatata cctgtggaac 1500gagcagcaac ctttgttttg gctacaagcc cttattccgc tggcagccct gattgttcta 1560tgcaactgtc tgagactctt accatgctgc tgtaaaacgt tggctttttt agccgtaatg 1620agcgtcggtg cccacactgt gagcgcgtac gaacacgtaa cagtgatccc gaacacggtg 1680ggagtaccgt ataagactct agtcaataga cctggctaca gccccatggt attggagatg 1740gaactactgt cagtcacttt ggagccaaca ctatcgcttg attacatcac gtgcgagtac 1800aaaaccgtca tcccgtctcc gtacgtgaag tgctgcggta cagcagagtg caaggacaaa 1860aacctacctg actacagctg taaggtcttc accggcgtct acccatttat gtggggcggc 1920gcctactgct tctgcgacgc tgaaaacacg cagttgagcg aagcacacgt ggagaagtcc 1980gaatcatgca aaacagaatt tgcatcagca tacagggctc ataccgcatc tgcatcagct 2040aagctccgcg tcctttacca aggaaataac atcactgtaa ctgcctatgc aaacggcgac 2100catgccgtca cagttaagga cgccaaattc attgtggggc caatgtcttc agcctggaca 2160cctttcgaca acaaaattgt ggtgtacaaa ggtgacgtct ataacatgga ctacccgccc 2220tttggcgcag gaagaccagg acaatttggc

gatatccaaa gtcgcacacc tgagagtaaa 2280gacgtctatg ctaatacaca actggtactg cagagaccgg ctgtgggtac ggtacacgtg 2340ccatactctc aggcaccatc tggctttaag tattggctaa aagaacgcgg ggcgtcgctg 2400cagcacacag caccatttgg ctgccaaata gcaacaaacc cggtaagagc ggtgaactgc 2460gccgtaggga acatgcccat ctccatcgac ataccggaag cggccttcac tagggtcgtc 2520gacgcgccct ctttaacgga catgtcgtgc gaggtaccag cctgcaccca ttcctcagac 2580tttgggggcg tcgccattat taaatatgca gccagcaaga aaggcaagtg tgcggtgcat 2640tcgatgacta acgccgtcac tattcgggaa gctgagatag aagttgaagg gaattctcag 2700ctgcaaatct ctttctcgac ggccttagcc agcgccgaat tccgcgtaca agtctgttct 2760acacaagtac actgtgcagc cgagtgccac cccccgaagg accacatagt caactacccg 2820gcgtcacata ccaccctcgg ggtccaggac atctccgcta cggcgatgtc atgggtgcag 2880aagatcacgg gaggtgtggg actggttgtt gctgttgccg cactgattct aatcgtggtg 2940ctatgcgtgt cgttcagcag gcac 296463747DNAChikungunya virus 6atggagttca tcccgacgca aactttctat aacagaaggt accaaccccg accctgggcc 60ccacgcccta caattcaagt aattagacct agaccacgtc cacagaggca ggctgggcaa 120ctcgcccagc tgatctccgc agtcaacaaa ttgaccatgc gcgcggtacc tcaacagaag 180cctcgcagaa atcggaaaaa caagaagcaa aggcagaaga agcaggcgcc gcaaaacgac 240ccaaagcaaa agaagcaacc accacaaaag aagccggctc aaaagaagaa gaaaccaggc 300cgtagggaga gaatgtgcat gaaaattgaa aatgattgca tcttcgaagt caagcatgaa 360ggcaaagtga tgggctacgc atgcctggtg ggggataaag taatgaaacc agcacatgtg 420aagggaacta tcgacaatgc cgatctggct aaactggcct ttaagcggtc gtctaaatac 480gatcttgaat gtgcacagat accggtgcac atgaagtctg atgcctcgaa gtttacccac 540gagaaacccg aggggtacta taactggcat cacggagcag tgcagtattc aggaggccgg 600ttcactatcc cgacgggtgc aggcaagccg ggagacagcg gcagaccgat cttcgacaac 660aaaggacggg tggtggccat cgtcctagga ggggccaacg aaggtgcccg cacggccctc 720tccgtggtga cgtggaacaa agacatcgtc acaaaaatta cccctgaggg agccgaagag 780tggagcctcg ccctcccggt cttgtgcctg ttggcaaaca ctacattccc ctgctctcag 840ccgccttgca caccctgctg ctacgaaaag gaaccggaaa gcaccttgcg catgcttgag 900gacaacgtga tgagacccgg atactaccag ctactaaaag catcgctgac ttgctctccc 960caccgccaaa gacgcagtac taaggacaat tttaatgtct ataaagccac aagaccatat 1020ctagctcatt gtcctgactg cggagaaggg cattcgtgcc acagccctat cgcattggag 1080cgcatcagaa atgaagcaac ggacggaacg ctgaaaatcc aggtctcttt gcagatcggg 1140ataaagacag atgacagcca cgattggacc aagctgcgct atatggatag ccatacgcca 1200gcggacgcgg agcgagccgg attgcttgta aggacttcag caccgtgcac gatcaccggg 1260accatgggac actttattct cgcccgatgc ccgaaaggag agacgctgac agtgggattt 1320acggacagca gaaagatcag ccacacatgc acacacccgt tccatcatga accacctgtg 1380ataggtaggg agaggttcca ctctcgacca caacatggta aagagttacc ttgcagcacg 1440tacgtgcaga gcaccgctgc cactgctgag gagatagagg tgcatatgcc cccagatact 1500cctgaccgca cgctgatgac gcagcagtct ggcaacgtga agatcacagt taatgggcag 1560acggtgcggt acaagtgcaa ctgcggtggc tcaaacgagg gactgacaac cacagacaaa 1620gtgatcaata actgcaaaat tgatcagtgc catgctgcag tcactaatca caagaattgg 1680caatacaact cccctttagt cccgcgcaac gctgaactcg gggaccgtaa aggaaagatc 1740cacatcccat tcccattggc aaacgtgact tgcagagtgc caaaagcaag aaaccctaca 1800gtaacttacg gaaaaaacca agtcaccatg ctgctgtatc ctgaccatcc gacactcttg 1860tcttaccgta acatgggaca ggaaccaaat taccacgagg agtgggtgac acacaagaag 1920gaggttacct tgaccgtgcc tactgagggt ctggaggtca cttggggcaa caacgaacca 1980tacaagtact ggccgcagat gtctacgaac ggtactgctc atggtcaccc acatgagata 2040atcttgtact attatgagct gtaccccact atgactgtag tcattgtgtc ggtggcctcg 2100ttcgtgcttc tgtcgatggt gggcacagca gtgggaatgt gtgtgtgcgc acggcgcaga 2160tgcattacac catatgaatt aacaccagga gccactgttc ccttcctgct cagcctgcta 2220tgctgcgtca gaacgaccaa ggcggccaca tattacgagg ctgcggcata tctatggaac 2280gaacagcagc ccctgttctg gttgcaggct cttatcccgc tggccgcctt gatcgtcctg 2340tgcaactgtc tgaaactctt gccatgctgc tgtaagaccc tggctttttt agccgtaatg 2400agcatcggtg cccacactgt gagcgcgtac gaacacgtaa cagtgatccc gaacacggtg 2460ggagtaccgt ataagactct tgtcaacaga ccgggttaca gccccatggt gttggagatg 2520gagctacaat cagtcacctt ggaaccaaca ctgtcacttg actacatcac gtgcgagtac 2580aaaactgtca tcccctcccc gtacgtgaag tgctgtggta cagcagagtg caaggacaag 2640agcctaccag actacagctg caaggtcttt actggagtct acccatttat gtggggcggc 2700gcctactgct tttgcgacgc cgaaaatacg caattgagcg aggcacatgt agagaaatct 2760gaatcttgca aaacagagtt tgcatcggcc tacagagccc acaccgcatc ggcgtcggcg 2820aagctccgcg tcctttacca aggaaacaac attaccgtag ctgcctacgc taacggtgac 2880catgccgtca cagtaaagga cgccaagttt gtcgtgggcc caatgtcctc cgcctggaca 2940ccttttgaca acaaaatcgt ggtgtacaaa ggcgacgtct acaacatgga ctacccacct 3000tttggcgcag gaagaccagg acaatttggt gacattcaaa gtcgtacacc ggaaagtaaa 3060gacgtttatg ccaacactca gttggtacta cagaggccag cagcaggcac ggtacatgta 3120ccatactctc aggcaccatc tggcttcaag tattggctga aggaacgagg agcatcgcta 3180cagcacacgg caccgttcgg ttgccagatt gcgacaaacc cggtaagagc tgtaaattgc 3240gctgtgggga acataccaat ttccatcgac ataccggatg cggcctttac tagggttgtc 3300gatgcaccct ctgtaacgga catgtcatgc gaagtaccag cctgcactca ctcctccgac 3360tttgggggcg tcgccatcat caaatacaca gctagcaaga aaggtaaatg tgcagtacat 3420tcgatgacca acgccgttac cattcgagaa gccgacgtag aagtagaggg gaactcccag 3480ctgcaaatat ccttctcaac agccctggca agcgccgagt ttcgcgtgca agtgtgctcc 3540acacaagtac actgcgcagc cgcatgccac cctccaaagg accacatagt caattaccca 3600gcatcacaca ccacccttgg ggtccaggat atatccacaa cggcaatgtc ttgggtgcag 3660aagattacgg gaggagtagg attaattgtt gctgttgctg ccttaatttt aattgtggtg 3720ctatgcgtgt cgtttagcag gcactaa 374773747DNAChikungunya virus 7atggagttca tcccaaccca aactttttac aataggaggt accagcctcg accctggact 60ccgcgcccta ctatccaagt catcaggccc agaccgcgcc ctcagaggca agctgggcaa 120cttgcccagc tgatctcagc agttaataaa ctgacaatgc gcgcggtacc acaacagaag 180ccacgcagga atcggaagaa taagaagcaa aagcaaaaac aacaggcgcc acaaaacaac 240acaaatcaaa agaagcagcc acctaaaaag aaaccggctc aaaagaaaaa gaagccgggc 300cgcagagaga ggatgtgcat gaaaatcgaa aatgattgta ttttcgaagt caagcacgaa 360ggtaaggtaa caggttacgc gtgcctggtg ggggacaaag taatgaaacc agcacacgta 420aaggggacca tcgataacgc ggacctggcc aaactggcct ttaagcggtc atctaagtat 480gaccttgaat gcgcgcagat acccgtgcac atgaagtccg acgcttcgaa gttcacccat 540gagaaaccgg aggggtacta caactggcac cacggagcag tacagtactc aggaggccgg 600ttcaccatcc ctacaggtgc tggcaaacca ggggacagcg gcagaccgat cttcgacaac 660aagggacgcg tggtggccat agtcttagga ggagctaatg aaggagcccg tacagccctc 720tcggtggtga cctggaataa agacattgtc actaaaatca cccccgaggg ggccgaagag 780tggagtcttg ccatcccagt tatgtgcctg ttggcaaaca ccacgttccc ctgctcccag 840cccccttgca cgccctgctg ctacgaaaag gaaccggagg aaaccctacg catgcttgag 900gacaacgtca tgagacctgg gtactatcag ctgctacaag catccttaac atgttctccc 960caccgccagc gacgcagcac caaggacaac ttcaatgtct ataaagccac aagaccatac 1020ttagctcact gtcccgactg tggagaaggg cactcgtgcc atagtcccgt agcactagaa 1080cgcatcagaa atgaagcgac agacgggacg ctgaaaatcc aggtctcctt gcaaatcgga 1140ataaagacgg atgacagcca cgattggacc aagctgcgtt atatggacaa ccacatgcca 1200gcagacgcag agagggcggg gctatttgta agaacatcag caccgtgtac gattactgga 1260acaatgggac acttcatcct ggcccgatgt ccaaaagggg aaactctgac ggtgggattc 1320actgacagta ggaagattag tcactcatgt acgcacccat ttcaccacga ccctcctgtg 1380ataggtcggg aaaaattcca ttcccgaccg cagcacggta aagagctacc ttgcagcacg 1440tacgtgcaga gcaccgccgc aactaccgag gagatagagg tacacatgcc cccagacacc 1500cctgatcgca cattaatgtc acaacagtcc ggcaacgtaa agatcacagt caatggccag 1560acggtgcggt acaagtgtaa ttgcggtggc tcaaatgaag gactaacaac tacagacaaa 1620gtgattaata actgcaaggt tgatcaatgt catgccgcgg tcaccaatca caaaaagtgg 1680cagtataact cccctctggt cccgcgtaat gctgaacttg gggaccgaaa aggaaaaatt 1740cacatcccgt ttccgctggc aaatgtaaca tgcagggtgc ctaaagcaag gaaccccacc 1800gtgacgtacg ggaaaaacca agtcatcatg ctactgtatc ctgaccaccc aacactcctg 1860tcctaccgga atatgggaga agaaccaaac tatcaagaag agtgggtgat gcataagaag 1920gaagtcgtgc taaccgtgcc gactgaaggg ctcgaggtca cgtggggcaa caacgagccg 1980tataagtatt ggccgcagtt atctacaaac ggtacagccc atggccaccc gcatgagata 2040attctgtatt attatgagct gtaccccact atgactgtag tagttgtgtc agtggccacg 2100ttcatactcc tgtcgatggt gggtatggca gcggggatgt gcatgtgtgc acgacgcaga 2160tgcatcacac cgtatgaact gacaccagga gctaccgtcc ctttcctgct tagcctaata 2220tgctgcatca gaacagctaa agcggccaca taccaagagg ctgcgatata cctgtggaac 2280gagcagcaac ctttgttttg gctacaagcc cttattccgc tggcagccct gattgttcta 2340tgcaactgtc tgagactctt accatgctgc tgtaaaacgt tggctttttt agccgtaatg 2400agcgtcggtg cccacactgt gagcgcgtac gaacacgtaa cagtgatccc gaacacggtg 2460ggagtaccgt ataagactct agtcaataga cctggctaca gccccatggt attggagatg 2520gaactactgt cagtcacttt ggagccaaca ctatcgcttg attacatcac gtgcgagtac 2580aaaaccgtca tcccgtctcc gtacgtgaag tgctgcggta cagcagagtg caaggacaaa 2640aacctacctg actacagctg taaggtcttc accggcgtct acccatttat gtggggcggc 2700gcctactgct tctgcgacgc tgaaaacacg cagttgagcg aagcacacgt ggagaagtcc 2760gaatcatgca aaacagaatt tgcatcagca tacagggctc ataccgcatc tgcatcagct 2820aagctccgcg tcctttacca aggaaataac atcactgtaa ctgcctatgc aaacggcgac 2880catgccgtca cagttaagga cgccaaattc attgtggggc caatgtcttc agcctggaca 2940cctttcgaca acaaaattgt ggtgtacaaa ggtgacgtct ataacatgga ctacccgccc 3000tttggcgcag gaagaccagg acaatttggc gatatccaaa gtcgcacacc tgagagtaaa 3060gacgtctatg ctaatacaca actggtactg cagagaccgg ctgtgggtac ggtacacgtg 3120ccatactctc aggcaccatc tggctttaag tattggctaa aagaacgcgg ggcgtcgctg 3180cagcacacag caccatttgg ctgccaaata gcaacaaacc cggtaagagc ggtgaactgc 3240gccgtaggga acatgcccat ctccatcgac ataccggaag cggccttcac tagggtcgtc 3300gacgcgccct ctttaacgga catgtcgtgc gaggtaccag cctgcaccca ttcctcagac 3360tttgggggcg tcgccattat taaatatgca gccagcaaga aaggcaagtg tgcggtgcat 3420tcgatgacta acgccgtcac tattcgggaa gctgagatag aagttgaagg gaattctcag 3480ctgcaaatct ctttctcgac ggccttagcc agcgccgaat tccgcgtaca agtctgttct 3540acacaagtac actgtgcagc cgagtgccac cccccgaagg accacatagt caactacccg 3600gcgtcacata ccaccctcgg ggtccaggac atctccgcta cggcgatgtc atgggtgcag 3660aagatcacgg gaggtgtggg actggttgtt gctgttgccg cactgattct aatcgtggtg 3720ctatgcgtgt cgttcagcag gcactaa 374788404DNAArtificial SequenceSynthetic vector VLP_CHI512 8gaattcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt 60ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg 120gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 180ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 240atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 300gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 360gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 420tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 480atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 540gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 600tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga 660gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 720agaagacacc gggaccgatc cagcctccgt taacggtgga gggcagtgta gtctgagcag 780tactcgttgc tgccgcgcgc gccaccagac ataatagctg acagactaac agactgttcc 840tttccatggg tcttttctgc agtcaccgtc gtcgacacgt gtgatcagat atcgcggccg 900ccaccatgga gttcatcccg acgcaaactt tctataacag aaggtaccaa ccccgaccct 960gggccccacg ccctacaatt caagtaatta gacctagacc acgtccacag aggcaggctg 1020ggcaactcgc ccagctgatc tccgcagtca acaaattgac catgcgcgcg gtacctcaac 1080agaagcctcg cagaaatcgg aaaaacaaga agcaaaggca gaagaagcag gcgccgcaaa 1140acgacccaaa gcaaaagaag caaccaccac aaaagaagcc ggctcaaaag aagaagaaac 1200caggccgtag ggagagaatg tgcatgaaaa ttgaaaatga ttgcatcttc gaagtcaagc 1260atgaaggcaa agtgatgggc tacgcatgcc tggtggggga taaagtaatg aaaccagcac 1320atgtgaaggg aactatcgac aatgccgatc tggctaaact ggcctttaag cggtcgtcta 1380aatacgatct tgaatgtgca cagataccgg tgcacatgaa gtctgatgcc tcgaagttta 1440cccacgagaa acccgagggg tactataact ggcatcacgg agcagtgcag tattcaggag 1500gccggttcac tatcccgacg ggtgcaggca agccgggaga cagcggcaga ccgatcttcg 1560acaacaaagg acgggtggtg gccatcgtcc taggaggggc caacgaaggt gcccgcacgg 1620ccctctccgt ggtgacgtgg aacaaagaca tcgtcacaaa aattacccct gagggagccg 1680aagagtggag cctcgccctc ccggtcttgt gcctgttggc aaacactaca ttcccctgct 1740ctcagccgcc ttgcacaccc tgctgctacg aaaaggaacc ggaaagcacc ttgcgcatgc 1800ttgaggacaa cgtgatgaga cccggatact accagctact aaaagcatcg ctgacttgct 1860ctccccaccg ccaaagacgc agtactaagg acaattttaa tgtctataaa gccacaagac 1920catatctagc tcattgtcct gactgcggag aagggcattc gtgccacagc cctatcgcat 1980tggagcgcat cagaaatgaa gcaacggacg gaacgctgaa aatccaggtc tctttgcaga 2040tcgggataaa gacagatgac agccacgatt ggaccaagct gcgctatatg gatagccata 2100cgccagcgga cgcggagcga gccggattgc ttgtaaggac ttcagcaccg tgcacgatca 2160ccgggaccat gggacacttt attctcgccc gatgcccgaa aggagagacg ctgacagtgg 2220gatttacgga cagcagaaag atcagccaca catgcacaca cccgttccat catgaaccac 2280ctgtgatagg tagggagagg ttccactctc gaccacaaca tggtaaagag ttaccttgca 2340gcacgtacgt gcagagcacc gctgccactg ctgaggagat agaggtgcat atgcccccag 2400atactcctga ccgcacgctg atgacgcagc agtccggagg atccaacgtg aagatcacag 2460ttaatgggca gacggtgcgg tacaagtgca actgcggtgg ctcaaacgag ggactgacaa 2520ccacagacaa agtgatcaat aactgcaaaa ttgatcagtg ccatgctgca gtcactaatc 2580acaagaattg gcaatacaac tcccctttag tcccgcgcaa cgctgaactc ggggaccgta 2640aaggaaagat ccacatccca ttcccattgg caaacgtgac ttgcagagtg ccaaaagcaa 2700gaaaccctac agtaacttac ggaaaaaacc aagtcaccat gctgctgtat cctgaccatc 2760cgacactctt gtcttaccgt aacatgggac aggaaccaaa ttaccacgag gagtgggtga 2820cacacaagaa ggaggttacc ttgaccgtgc ctactgaggg tctggaggtc acttggggca 2880acaacgaacc atacaagtac tggccgcaga tgtctacgaa cggtactgct catggtcacc 2940cacatgagat aatcttgtac tattatgagc tgtaccccac tatgactgta gtcattgtgt 3000cggtggcctc gttcgtgctt ctgtcgatgg tgggcacagc agtgggaatg tgtgtgtgcg 3060cacggcgcag atgcattaca ccatatgaat taacaccagg agccactgtt cccttcctgc 3120tcagcctgct atgctgcgtc agaacgacca aggcggccac atattacgag gctgcggcat 3180atctatggaa cgaacagcag cccctgttct ggttgcaggc tcttatcccg ctggccgcct 3240tgatcgtcct gtgcaactgt ctgaaactct tgccatgctg ctgtaagacc ctggcttttt 3300tagccgtaat gagcatcggt gcccacactg tgagcgcgta cgaacacgta acagtgatcc 3360cgaacacggt gggagtaccg tataagactc ttgtcaacag accgggttac agccccatgg 3420tgttggagat ggagctacaa tcagtcacct tggaaccaac actgtcactt gactacatca 3480cgtgcgagta caaaactgtc atcccctccc cgtacgtgaa gtgctgtggt acagcagagt 3540gcaaggacaa gagcctacca gactacagct gcaaggtctt tactggagtc tacccattta 3600tgtggggcgg cgcctactgc ttttgcgacg ccgaaaatac gcaattgagc gaggcacatg 3660tagagaaatc tgaatcttgc aaaacagagt ttgcatcggc ctacagagcc cacaccgcat 3720cggcgtcggc gaagctccgc gtcctttacc aaggaaacaa cattaccgta gctgcctacg 3780ctaacggtga ccatgccgtc acagtaaagg acgccaagtt tgtcgtgggc ccaatgtcct 3840ccgcctggac accttttgac aacaaaatcg tggtgtacaa aggcgacgtc tacaacatgg 3900actacccacc ttttggcgca ggaagaccag gacaatttgg tgacattcaa agtcgtacac 3960cggaaagtaa agacgtttat gccaacactc agttggtact acagaggcca gcagcaggca 4020cggtacatgt accatactct caggcaccat ctggcttcaa gtattggctg aaggaacgag 4080gagcatcgct acagcacacg gcaccgttcg gttgccagat tgcgacaaac ccggtaagag 4140ctgtaaattg cgctgtgggg aacataccaa tttccatcga cataccggat gcggccttta 4200ctagggttgt cgatgcaccc tctgtaacgg acatgtcatg cgaagtacca gcctgcactc 4260actcctccga ctttgggggc gtcgccatca tcaaatacac agctagcaag aaaggtaaat 4320gtgcagtaca ttcgatgacc aacgccgtta ccattcgaga agccgacgta gaagtagagg 4380ggaactccca gctgcaaata tccttctcaa cagccctggc aagcgccgag tttcgcgtgc 4440aagtgtgctc cacacaagta cactgcgcag ccgcatgcca ccctccaaag gaccacatag 4500tcaattaccc agcatcacac accacccttg gggtccagga tatatccaca acggcaatgt 4560cttgggtgca gaagattacg ggaggagtag gattaattgt tgctgttgct gccttaattt 4620taattgtggt gctatgcgtg tcgtttagca ggcactaagg atctagatct gctgtgcctt 4680ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg 4740ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt 4800gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca 4860atagcaggca tgctggggat gcggtgggct ctatgggtac ccaggtgctg aagaattgac 4920ccggttcctc ctgggccaga aagaagcagg cacatcccct tctctgtgac acaccctgtc 4980cacgcccctg gttcttagtt ccagccccac tcataggaca ctcatagctc aggagggctc 5040cgccttcaat cccacccgct aaagtacttg gagcggtctc tccctccctc atcagcccac 5100caaaccaaac ctagcctcca agagtgggaa gaaattaaag caagataggc tattaagtgc 5160agagggagag aaaatgcctc caacatgtga ggaagtaatg agagaaatca tagaatttta 5220aggccatgat ttaaggccat catggcctaa gcttgaaagg agataggatc aaagcttggc 5280gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 5340catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 5400attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 5460ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 5520ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 5580aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 5640aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 5700gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 5760gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 5820tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 5880ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 5940ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 6000tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 6060tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 6120ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 6180aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 6240ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 6300tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 6360atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 6420aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 6480ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 6540tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg 6600ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg

agcgcagaag 6660tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 6720aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 6780gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 6840tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 6900cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 6960tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 7020ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 7080cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 7140actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 7200ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 7260aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 7320ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 7380atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 7440tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag 7500gccctttcgg gtcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc 7560gttgacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc 7620gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt 7680actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt 7740ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc 7800aaaagaatag cccgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt 7860aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact 7920acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg 7980gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag 8040aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac 8100gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtactatgg 8160ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc 8220aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct 8280tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg 8340ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattcca tggtctcaac 8400tttc 840498410DNAArtificial SequenceSynthetic vector VLP_CHI 532 9gaattcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt 60ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg 120gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 180ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 240atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 300gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 360gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 420tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 480atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 540gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 600tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga 660gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 720agaagacacc gggaccgatc cagcctccgt taacggtgga gggcagtgta gtctgagcag 780tactcgttgc tgccgcgcgc gccaccagac ataatagctg acagactaac agactgttcc 840tttccatggg tcttttctgc agtcaccgtc gtcgacacgt gtgatcagat atcgcggccg 900ccaccatgga gttcatcccg acgcaaactt tctataacag aaggtaccaa ccccgaccct 960gggccccacg ccctacaatt caagtaatta gacctagacc acgtccacag aggcaggctg 1020ggcaactcgc ccagctgatc tccgcagtca acaaattgac catgcgcgcg gtacctcaac 1080agaagcctcg cagaaatcgg aaaaacaaga agcaaaggca gaagaagcag gcgccgcaaa 1140acgacccaaa gcaaaagaag caaccaccac aaaagaagcc ggctcaaaag aagaagaaac 1200caggccgtag ggagagaatg tgcatgaaaa ttgaaaatga ttgcatcttc gaagtcaagc 1260atgaaggcaa agtgatgggc tacgcatgcc tggtggggga taaagtaatg aaaccagcac 1320atgtgaaggg aactatcgac aatgccgatc tggctaaact ggcctttaag cggtcgtcta 1380aatacgatct tgaatgtgca cagataccgg tgcacatgaa gtctgatgcc tcgaagttta 1440cccacgagaa acccgagggg tactataact ggcatcacgg agcagtgcag tattcaggag 1500gccggttcac tatcccgacg ggtgcaggca agccgggaga cagcggcaga ccgatcttcg 1560acaacaaagg acgggtggtg gccatcgtcc taggaggggc caacgaaggt gcccgcacgg 1620ccctctccgt ggtgacgtgg aacaaagaca tcgtcacaaa aattacccct gagggagccg 1680aagagtggag cctcgccctc ccggtcttgt gcctgttggc aaacactaca ttcccctgct 1740ctcagccgcc ttgcacaccc tgctgctacg aaaaggaacc ggaaagcacc ttgcgcatgc 1800ttgaggacaa cgtgatgaga cccggatact accagctact aaaagcatcg ctgacttgct 1860ctccccaccg ccaaagacgc agtactaagg acaattttaa tgtctataaa gccacaagac 1920catatctagc tcattgtcct gactgcggag aagggcattc gtgccacagc cctatcgcat 1980tggagcgcat cagaaatgaa gcaacggacg gaacgctgaa aatccaggtc tctttgcaga 2040tcgggataaa gacagatgac agccacgatt ggaccaagct gcgctatatg gatagccata 2100cgccagcgga cgcggagcga gccggattgc ttgtaaggac ttcagcaccg tgcacgatca 2160ccgggaccat gggacacttt attctcgccc gatgcccgaa aggagagacg ctgacagtgg 2220gatttacgga cagcagaaag atcagccaca catgcacaca cccgttccat catgaaccac 2280ctgtgatagg tagggagagg ttccactctc gaccacaaca tggtaaagag ttaccttgca 2340gcacgtacgt gcagagcacc gctgccactg ctgaggagat agaggtgcat atgcccccag 2400atactcctga ccgcacgctg atgacgcagc agtctggcaa cgtgaagatc acagttaatg 2460ggcagacggt gcggtacaag tgcaactgcg gtggctccgg aagtggatcc aacgagggac 2520tgacaaccac agacaaagtg atcaataact gcaaaattga tcagtgccat gctgcagtca 2580ctaatcacaa gaattggcaa tacaactccc ctttagtccc gcgcaacgct gaactcgggg 2640accgtaaagg aaagatccac atcccattcc cattggcaaa cgtgacttgc agagtgccaa 2700aagcaagaaa ccctacagta acttacggaa aaaaccaagt caccatgctg ctgtatcctg 2760accatccgac actcttgtct taccgtaaca tgggacagga accaaattac cacgaggagt 2820gggtgacaca caagaaggag gttaccttga ccgtgcctac tgagggtctg gaggtcactt 2880ggggcaacaa cgaaccatac aagtactggc cgcagatgtc tacgaacggt actgctcatg 2940gtcacccaca tgagataatc ttgtactatt atgagctgta ccccactatg actgtagtca 3000ttgtgtcggt ggcctcgttc gtgcttctgt cgatggtggg cacagcagtg ggaatgtgtg 3060tgtgcgcacg gcgcagatgc attacaccat atgaattaac accaggagcc actgttccct 3120tcctgctcag cctgctatgc tgcgtcagaa cgaccaaggc ggccacatat tacgaggctg 3180cggcatatct atggaacgaa cagcagcccc tgttctggtt gcaggctctt atcccgctgg 3240ccgccttgat cgtcctgtgc aactgtctga aactcttgcc atgctgctgt aagaccctgg 3300cttttttagc cgtaatgagc atcggtgccc acactgtgag cgcgtacgaa cacgtaacag 3360tgatcccgaa cacggtggga gtaccgtata agactcttgt caacagaccg ggttacagcc 3420ccatggtgtt ggagatggag ctacaatcag tcaccttgga accaacactg tcacttgact 3480acatcacgtg cgagtacaaa actgtcatcc cctccccgta cgtgaagtgc tgtggtacag 3540cagagtgcaa ggacaagagc ctaccagact acagctgcaa ggtctttact ggagtctacc 3600catttatgtg gggcggcgcc tactgctttt gcgacgccga aaatacgcaa ttgagcgagg 3660cacatgtaga gaaatctgaa tcttgcaaaa cagagtttgc atcggcctac agagcccaca 3720ccgcatcggc gtcggcgaag ctccgcgtcc tttaccaagg aaacaacatt accgtagctg 3780cctacgctaa cggtgaccat gccgtcacag taaaggacgc caagtttgtc gtgggcccaa 3840tgtcctccgc ctggacacct tttgacaaca aaatcgtggt gtacaaaggc gacgtctaca 3900acatggacta cccacctttt ggcgcaggaa gaccaggaca atttggtgac attcaaagtc 3960gtacaccgga aagtaaagac gtttatgcca acactcagtt ggtactacag aggccagcag 4020caggcacggt acatgtacca tactctcagg caccatctgg cttcaagtat tggctgaagg 4080aacgaggagc atcgctacag cacacggcac cgttcggttg ccagattgcg acaaacccgg 4140taagagctgt aaattgcgct gtggggaaca taccaatttc catcgacata ccggatgcgg 4200cctttactag ggttgtcgat gcaccctctg taacggacat gtcatgcgaa gtaccagcct 4260gcactcactc ctccgacttt gggggcgtcg ccatcatcaa atacacagct agcaagaaag 4320gtaaatgtgc agtacattcg atgaccaacg ccgttaccat tcgagaagcc gacgtagaag 4380tagaggggaa ctcccagctg caaatatcct tctcaacagc cctggcaagc gccgagtttc 4440gcgtgcaagt gtgctccaca caagtacact gcgcagccgc atgccaccct ccaaaggacc 4500acatagtcaa ttacccagca tcacacacca cccttggggt ccaggatata tccacaacgg 4560caatgtcttg ggtgcagaag attacgggag gagtaggatt aattgttgct gttgctgcct 4620taattttaat tgtggtgcta tgcgtgtcgt ttagcaggca ctaaggatct agatctgctg 4680tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg 4740aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga 4800gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg 4860aagacaatag caggcatgct ggggatgcgg tgggctctat gggtacccag gtgctgaaga 4920attgacccgg ttcctcctgg gccagaaaga agcaggcaca tccccttctc tgtgacacac 4980cctgtccacg cccctggttc ttagttccag ccccactcat aggacactca tagctcagga 5040gggctccgcc ttcaatccca cccgctaaag tacttggagc ggtctctccc tccctcatca 5100gcccaccaaa ccaaacctag cctccaagag tgggaagaaa ttaaagcaag ataggctatt 5160aagtgcagag ggagagaaaa tgcctccaac atgtgaggaa gtaatgagag aaatcataga 5220attttaaggc catgatttaa ggccatcatg gcctaagctt gaaaggagat aggatccaag 5280cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc 5340acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta 5400actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca 5460gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 5520cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 5580tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 5640gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 5700ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 5760aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 5820tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 5880ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 5940gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 6000tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 6060caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 6120ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt 6180cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 6240ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 6300cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 6360gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 6420aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 6480acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 6540gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 6600accacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 6660cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 6720tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 6780cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 6840gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 6900cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 6960ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 7020gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 7080taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 7140gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 7200acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 7260aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 7320cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 7380atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 7440gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat 7500cacgaggccc tttcgggtcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca 7560gctcccgttg acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca 7620gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca 7680gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt 7740aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta 7800taaatcaaaa gaatagcccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc 7860actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg 7920cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact 7980aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt 8040ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc 8100ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta 8160ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac 8220cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg 8280gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg 8340gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attccatggt 8400ctcaactttc 8410108431DNAArtificial SequenceSynthetic vector VLP_VEEV VLP 518 10gaattcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt 60ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg 120gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 180ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 240atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 300gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 360gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 420tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 480atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 540gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 600tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga 660gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 720agaagacacc gggaccgatc cagcctccgt taacggtgga gggcagtgta gtctgagcag 780tactcgttgc tgccgcgcgc gccaccagac ataatagctg acagactaac agactgttcc 840tttccatggg tcttttctgc agtcaccgtc gtcgacacgt gtgatcagat atcgcggccg 900ccaccatgtt cccgttccag ccaatgtatc cgatgcagcc aatgccctat cgcaacccgt 960tcgcggcccc gcgcaggccc tggttcccca gaaccgaccc ttttctggcg atgcaggtgc 1020aggaattaac ccgctcgatg gctaacctga cgttcaagca acgccgggac gcgccacctg 1080aggggccatc cgctaataaa ccgaagaagg aggcctcgca aaaacagaaa gggggaggcc 1140aagggaagaa gaagaagaac caagggaaga agaaggctaa gacagggccg cctaatccga 1200aggcacagaa tggaaacaag aagaagacca acaagaaacc aggcaagaga cagcgcatgg 1260tcatgaaatt ggaatctgac aagacgttcc caatcatgtt ggaagggaag ataaacggct 1320acgcttgtgt ggtcggaggg aagttattca ggccgatgca tgtggaaggc aagatcgaca 1380acgacgttct ggccgcgctt aagacgaaga aagcatccaa atacgatctt gagtatgcag 1440atgtgccaca gaacatgcgg gccgatacat tcaaatacac ccatgagaaa ccccaaggct 1500attacagctg gcatcatgga gcagtccaat atgaaaatgg gcgtttcacg gtgccgaaag 1560gagttggggc caagggagac agcggacgac ccattctgga taaccaggga cgggtggtcg 1620ctattgtgct gggaggtgtg aatgaaggat ctaggacagc cctttcagtc gtcatgtgga 1680acgagaaggg agttaccgtg aagtatactc cagagaactg cgagcaatgg tcactagtga 1740ccaccatgtg tctgctcgcc aatgtgacgt tcccatgtgc tcaaccacca atttgctacg 1800acagaaaacc agcagagact ttggccatgc tcagcgttaa cgttgacaac ccgggctacg 1860atgagctgct ggaagcagct gttaagtgcc ccggaaggaa aaggagatcc accgaggagc 1920tgtttaatga gtataagcta acgcgccctt acatggccag atgcatcaga tgtgcagttg 1980ggagctgcca tagtccaata gcaatcgagg cagtaaagag cgacgggcac gacggttatg 2040ttagacttca gacttcctcg cagtatggcc tggattcctc cggcaactta aagggcagga 2100ccatgcggta tgacatgcac gggaccatta aagagatacc actacatcaa gtgtcactct 2160atacatctcg cccgtgtcac attgtggatg ggcacggtta tttcctgctt gccaggtgcc 2220cggcagggga ctccatcacc atggaattta agaaagattc cgtcagacac tcctgctcgg 2280tgccgtatga agtgaaattt aatcctgtag gcagagaact ctatactcat cccccagaac 2340acggagtaga gcaagcgtgc caagtctacg cacatgatgc acagaacaga ggagcttatg 2400tcgagatgca cctcccgggc tcagaagtgg acagcagttt ggtttccttg agcggctccg 2460gaggatccag ttcagtcacc gtgacacctc ctgatgggac tagcgccctg gtggaatgcg 2520agtgtggcgg cacaaagatc tccgagacca tcaacaagac aaaacagttc agccagtgca 2580caaagaagga gcagtgcaga gcatatcggc tgcagaacga taagtgggtg tataattctg 2640acaaactgcc caaagcagcg ggagccacct taaaaggaaa actgcatgtc ccattcttgc 2700tggcagacgg caaatgcacc gtgcctctag caccagaacc tatgataacc ttcggtttca 2760gatcagtgtc actgaaactg caccctaaga atcccacata tctaatcacc cgccaacttg 2820ctgatgagcc tcactacacg cacgagctca tatctgaacc agctgttagg aattttaccg 2880tcaccgaaaa agggtgggag tttgtatggg gaaaccaccc gccgaaaagg ttttgggcac 2940aggaaacagc acccggaaat ccacatgggc taccgcacga ggtgataact cattattacc 3000acagataccc tatgtccacc atcctgggtt tgtcaatttg tgccgccatt gcaaccgttt 3060ccgttgcagc gtctacctgg ctgttttgca gatcaagagt tgcgtgccta actccttacc 3120ggctaacacc taacgctagg ataccatttt gtctggctgt gctttgctgc gcccgcactg 3180cccgggccga gaccacctgg gagtccttgg atcacctatg gaacaataac caacagatgt 3240tctggattca attgctgatc cctctggccg ccttgatcgt agtgactcgc ctgctcaggt 3300gcgtgtgctg tgtcgtgcct tttttagtca tggccggcgc cgcaggcgcc ggcgcctacg 3360agcacgcgac cacgatgccg agccaagcgg gaatctcgta taacactata gtcaacagag 3420caggctacgc accactccct atcagcataa caccaacaaa gatcaagctg atacctacag 3480tgaacttgga gtacgtcacc tgccactaca aaacaggaat ggattcacca gccatcaaat 3540gctgcggatc tcaggaatgc actccaactt acaggcctga tgaacagtgc aaagtcttca 3600caggggttta cccgttcatg tggggtggtg catattgctt ttgcgacact gagaacaccc 3660aagtcagcaa ggcctacgta atgaaatctg acgactgcct tgcggatcat gctgaagcat 3720ataaagcgca cacagcctca gtgcaggcgt tcctcaacat cacagtggga gaacactcta 3780ttgtgactac cgtgtatgtg aatggagaaa ctcctgtgaa tttcaatggg gtcaaaataa 3840ctgcaggtcc gctttccaca gcttggacac cctttgatcg caaaatcgtg cagtatgccg 3900gggagatcta taattatgat tttcctgagt atggggcagg acaaccagga gcatttggag 3960atatacaatc cagaacagtc tcaagctctg atctgtatgc caataccaac ctagtgctgc 4020agagacccaa agcaggagcg atccacgtgc catacactca ggcaccttcg ggttttgagc 4080aatggaagaa agataaagct ccatcattga aatttaccgc ccctttcgga tgcgaaatat 4140atacaaaccc cattcgcgcc gaaaactgtg ctgtagggtc aattccatta gcctttgaca 4200ttcccgacgc cttgttcacc agggtgtcag aaacaccgac actttcagcg gccgaatgca 4260ctcttaacga gtgcgtgtat tcttccgact ttggtgggat cgccacggtc aagtactcgg 4320ccagcaagtc aggcaagtgc gcagtccatg tgccatcagg gactgctacc ctaaaagaag 4380cagcagtcga gctaaccgag caagggtcgg cgactatcca tttctcgacc gcaaatatcc 4440acccggagtt caggctccaa atatgcacat catatgttac gtgcaaaggt gattgtcacc 4500ccccgaaaga ccatattgtg acacaccctc agtatcacgc ccaaacattt acagccgcgg 4560tgtcaaaaac cgcgtggacg tggttaacat ccctgctggg aggatcagcc gtaattatta 4620taattggctt ggtgctggct actattgtgg ccatgtacgt gctgaccaac cagaaacata

4680attaaggatc tagatctgct gtgccttcta gttgccagcc atctgttgtt tgcccctccc 4740ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg 4800aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg 4860acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta 4920tgggtaccca ggtgctgaag aattgacccg gttcctcctg ggccagaaag aagcaggcac 4980atccccttct ctgtgacaca ccctgtccac gcccctggtt cttagttcca gccccactca 5040taggacactc atagctcagg agggctccgc cttcaatccc acccgctaaa gtacttggag 5100cggtctctcc ctccctcatc agcccaccaa accaaaccta gcctccaaga gtgggaagaa 5160attaaagcaa gataggctat taagtgcaga gggagagaaa atgcctccaa catgtgagga 5220agtaatgaga gaaatcatag aattttaagg ccatgattta aggccatcat ggcctaagct 5280tgaaaggaga taggatcaaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 5340ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 5400gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 5460gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 5520tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 5580gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 5640ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 5700ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 5760acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 5820tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 5880ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 5940ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 6000ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 6060actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 6120gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 6180tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 6240caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 6300atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 6360acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 6420ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 6480ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 6540tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 6600tgctgcaatg ataccgcgag aaccacgctc accggctcca gatttatcag caataaacca 6660gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 6720tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 6780tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 6840ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 6900tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 6960ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 7020gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 7080ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 7140cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 7200ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 7260ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 7320gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 7380ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 7440gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt 7500aacctataaa aataggcgta tcacgaggcc ctttcgggtc gcgcgtttcg gtgatgacgg 7560tgaaaacctc tgacacatgc agctcccgtt gacggtcaca gcttgtctgt aagcggatgc 7620cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct 7680taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa 7740tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc 7800cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagggt tgagtgttgt 7860tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa 7920aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg 7980gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg 8040acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc 8100tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa 8160tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac 8220agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt 8280tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt 8340gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg 8400acggccagtg aattccatgg tctcaacttt c 8431118431DNAArtificial SequenceSynthetic vector VLP_VEEV VLP 519 11gaattcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt 60ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg 120gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 180ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 240atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 300gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 360gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 420tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 480atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 540gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 600tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga 660gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 720agaagacacc gggaccgatc cagcctccgt taacggtgga gggcagtgta gtctgagcag 780tactcgttgc tgccgcgcgc gccaccagac ataatagctg acagactaac agactgttcc 840tttccatggg tcttttctgc agtcaccgtc gtcgacacgt gtgatcagat atcgcggccg 900ccaccatgtt cccgttccag ccaatgtatc cgatgcagcc aatgccctat cgcaacccgt 960tcgcggcccc gcgcaggccc tggttcccca gaaccgaccc ttttctggcg atgcaggtgc 1020aggaattaac ccgctcgatg gctaacctga cgttcaagca acgccgggac gcgccacctg 1080aggggccatc cgctaataaa ccgaagaagg aggcctcgca aaaacagaaa gggggaggcc 1140aagggaagaa gaagaagaac caagggaaga agaaggctaa gacagggccg cctaatccga 1200aggcacagaa tggaaacaag aagaagacca acaagaaacc aggcaagaga cagcgcatgg 1260tcatgaaatt ggaatctgac aagacgttcc caatcatgtt ggaagggaag ataaacggct 1320acgcttgtgt ggtcggaggg aagttattca ggccgatgca tgtggaaggc aagatcgaca 1380acgacgttct ggccgcgctt aagacgaaga aagcatccaa atacgatctt gagtatgcag 1440atgtgccaca gaacatgcgg gccgatacat tcaaatacac ccatgagaaa ccccaaggct 1500attacagctg gcatcatgga gcagtccaat atgaaaatgg gcgtttcacg gtgccgaaag 1560gagttggggc caagggagac agcggacgac ccattctgga taaccaggga cgggtggtcg 1620ctattgtgct gggaggtgtg aatgaaggat ctaggacagc cctttcagtc gtcatgtgga 1680acgagaaggg agttaccgtg aagtatactc cagagaactg cgagcaatgg tcactagtga 1740ccaccatgtg tctgctcgcc aatgtgacgt tcccatgtgc tcaaccacca atttgctacg 1800acagaaaacc agcagagact ttggccatgc tcagcgttaa cgttgacaac ccgggctacg 1860atgagctgct ggaagcagct gttaagtgcc ccggaaggaa aaggagatcc accgaggagc 1920tgtttaatga gtataagcta acgcgccctt acatggccag atgcatcaga tgtgcagttg 1980ggagctgcca tagtccaata gcaatcgagg cagtaaagag cgacgggcac gacggttatg 2040ttagacttca gacttcctcg cagtatggcc tggattcctc cggcaactta aagggcagga 2100ccatgcggta tgacatgcac gggaccatta aagagatacc actacatcaa gtgtcactct 2160atacatctcg cccgtgtcac attgtggatg ggcacggtta tttcctgctt gccaggtgcc 2220cggcagggga ctccatcacc atggaattta agaaagattc cgtcagacac tcctgctcgg 2280tgccgtatga agtgaaattt aatcctgtag gcagagaact ctatactcat cccccagaac 2340acggagtaga gcaagcgtgc caagtctacg cacatgatgc acagaacaga ggagcttatg 2400tcgagatgca cctcccgggc tcagaagtgg acagcagttt ggtttccttg agcggcagtt 2460ccggaggatc ctcagtcacc gtgacacctc ctgatgggac tagcgccctg gtggaatgcg 2520agtgtggcgg cacaaagatc tccgagacca tcaacaagac aaaacagttc agccagtgca 2580caaagaagga gcagtgcaga gcatatcggc tgcagaacga taagtgggtg tataattctg 2640acaaactgcc caaagcagcg ggagccacct taaaaggaaa actgcatgtc ccattcttgc 2700tggcagacgg caaatgcacc gtgcctctag caccagaacc tatgataacc ttcggtttca 2760gatcagtgtc actgaaactg caccctaaga atcccacata tctaatcacc cgccaacttg 2820ctgatgagcc tcactacacg cacgagctca tatctgaacc agctgttagg aattttaccg 2880tcaccgaaaa agggtgggag tttgtatggg gaaaccaccc gccgaaaagg ttttgggcac 2940aggaaacagc acccggaaat ccacatgggc taccgcacga ggtgataact cattattacc 3000acagataccc tatgtccacc atcctgggtt tgtcaatttg tgccgccatt gcaaccgttt 3060ccgttgcagc gtctacctgg ctgttttgca gatcaagagt tgcgtgccta actccttacc 3120ggctaacacc taacgctagg ataccatttt gtctggctgt gctttgctgc gcccgcactg 3180cccgggccga gaccacctgg gagtccttgg atcacctatg gaacaataac caacagatgt 3240tctggattca attgctgatc cctctggccg ccttgatcgt agtgactcgc ctgctcaggt 3300gcgtgtgctg tgtcgtgcct tttttagtca tggccggcgc cgcaggcgcc ggcgcctacg 3360agcacgcgac cacgatgccg agccaagcgg gaatctcgta taacactata gtcaacagag 3420caggctacgc accactccct atcagcataa caccaacaaa gatcaagctg atacctacag 3480tgaacttgga gtacgtcacc tgccactaca aaacaggaat ggattcacca gccatcaaat 3540gctgcggatc tcaggaatgc actccaactt acaggcctga tgaacagtgc aaagtcttca 3600caggggttta cccgttcatg tggggtggtg catattgctt ttgcgacact gagaacaccc 3660aagtcagcaa ggcctacgta atgaaatctg acgactgcct tgcggatcat gctgaagcat 3720ataaagcgca cacagcctca gtgcaggcgt tcctcaacat cacagtggga gaacactcta 3780ttgtgactac cgtgtatgtg aatggagaaa ctcctgtgaa tttcaatggg gtcaaaataa 3840ctgcaggtcc gctttccaca gcttggacac cctttgatcg caaaatcgtg cagtatgccg 3900gggagatcta taattatgat tttcctgagt atggggcagg acaaccagga gcatttggag 3960atatacaatc cagaacagtc tcaagctctg atctgtatgc caataccaac ctagtgctgc 4020agagacccaa agcaggagcg atccacgtgc catacactca ggcaccttcg ggttttgagc 4080aatggaagaa agataaagct ccatcattga aatttaccgc ccctttcgga tgcgaaatat 4140atacaaaccc cattcgcgcc gaaaactgtg ctgtagggtc aattccatta gcctttgaca 4200ttcccgacgc cttgttcacc agggtgtcag aaacaccgac actttcagcg gccgaatgca 4260ctcttaacga gtgcgtgtat tcttccgact ttggtgggat cgccacggtc aagtactcgg 4320ccagcaagtc aggcaagtgc gcagtccatg tgccatcagg gactgctacc ctaaaagaag 4380cagcagtcga gctaaccgag caagggtcgg cgactatcca tttctcgacc gcaaatatcc 4440acccggagtt caggctccaa atatgcacat catatgttac gtgcaaaggt gattgtcacc 4500ccccgaaaga ccatattgtg acacaccctc agtatcacgc ccaaacattt acagccgcgg 4560tgtcaaaaac cgcgtggacg tggttaacat ccctgctggg aggatcagcc gtaattatta 4620taattggctt ggtgctggct actattgtgg ccatgtacgt gctgaccaac cagaaacata 4680attaaggatc tagatctgct gtgccttcta gttgccagcc atctgttgtt tgcccctccc 4740ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg 4800aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg 4860acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta 4920tgggtaccca ggtgctgaag aattgacccg gttcctcctg ggccagaaag aagcaggcac 4980atccccttct ctgtgacaca ccctgtccac gcccctggtt cttagttcca gccccactca 5040taggacactc atagctcagg agggctccgc cttcaatccc acccgctaaa gtacttggag 5100cggtctctcc ctccctcatc agcccaccaa accaaaccta gcctccaaga gtgggaagaa 5160attaaagcaa gataggctat taagtgcaga gggagagaaa atgcctccaa catgtgagga 5220agtaatgaga gaaatcatag aattttaagg ccatgattta aggccatcat ggcctaagct 5280tgaaaggaga taggatcaaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 5340ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 5400gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 5460gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 5520tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 5580gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 5640ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 5700ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 5760acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 5820tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 5880ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 5940ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 6000ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 6060actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 6120gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 6180tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 6240caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 6300atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 6360acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 6420ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 6480ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 6540tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 6600tgctgcaatg ataccgcgag aaccacgctc accggctcca gatttatcag caataaacca 6660gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 6720tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 6780tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 6840ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 6900tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 6960ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 7020gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 7080ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 7140cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 7200ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 7260ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 7320gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 7380ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 7440gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt 7500aacctataaa aataggcgta tcacgaggcc ctttcgggtc gcgcgtttcg gtgatgacgg 7560tgaaaacctc tgacacatgc agctcccgtt gacggtcaca gcttgtctgt aagcggatgc 7620cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct 7680taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa 7740tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc 7800cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagggt tgagtgttgt 7860tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa 7920aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg 7980gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg 8040acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc 8100tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa 8160tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac 8220agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt 8280tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt 8340gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg 8400acggccagtg aattccatgg tctcaacttt c 8431128431DNAArtificial SequenceSynthetic vector VLP_VEEV VLP 538 12gaattcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt 60ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg 120gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 180ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 240atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 300gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 360gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 420tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 480atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 540gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 600tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga 660gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 720agaagacacc gggaccgatc cagcctccgt taacggtgga gggcagtgta gtctgagcag 780tactcgttgc tgccgcgcgc gccaccagac ataatagctg acagactaac agactgttcc 840tttccatggg tcttttctgc agtcaccgtc gtcgacacgt gtgatcagat atcgcggccg 900ccaccatgtt cccgttccag ccaatgtatc cgatgcagcc aatgccctat cgcaacccgt 960tcgcggcccc gcgcaggccc tggttcccca gaaccgaccc ttttctggcg atgcaggtgc 1020aggaattaac ccgctcgatg gctaacctga cgttcaagca acgccgggac gcgccacctg 1080aggggccatc cgctaataaa ccgaagaagg aggcctcgca aaaacagaaa gggggaggcc 1140aagggaagaa gaagaagaac caagggaaga agaaggctaa gacagggccg cctaatccga 1200aggcacagaa tggaaacaag aagaagacca acaagaaacc aggcaagaga cagcgcatgg 1260tcatgaaatt ggaatctgac aagacgttcc caatcatgtt ggaagggaag ataaacggct 1320acgcttgtgt ggtcggaggg aagttattca ggccgatgca tgtggaaggc aagatcgaca 1380acgacgttct ggccgcgctt aagacgaaga aagcatccaa atacgatctt gagtatgcag 1440atgtgccaca gaacatgcgg gccgatacat tcaaatacac ccatgagaaa ccccaaggct 1500attacagctg gcatcatgga gcagtccaat atgaaaatgg gcgtttcacg gtgccgaaag 1560gagttggggc caagggagac agcggacgac ccattctgga taaccaggga cgggtggtcg 1620ctattgtgct gggaggtgtg aatgaaggat ctaggacagc cctttcagtc gtcatgtgga 1680acgagaaggg agttaccgtg aagtatactc cagagaactg cgagcaatgg tcactagtga 1740ccaccatgtg tctgctcgcc aatgtgacgt tcccatgtgc tcaaccacca atttgctacg 1800acagaaaacc agcagagact ttggccatgc tcagcgttaa cgttgacaac ccgggctacg 1860atgagctgct ggaagcagct gttaagtgcc ccggaaggaa aaggagatcc accgaggagc 1920tgtttaatga gtataagcta acgcgccctt acatggccag atgcatcaga tgtgcagttg 1980ggagctgcca tagtccaata gcaatcgagg cagtaaagag cgacgggcac gacggttatg 2040ttagacttca gacttcctcg cagtatggcc tggattcctc cggcaactta aagggcagga 2100ccatgcggta tgacatgcac gggaccatta aagagatacc actacatcaa gtgtcactct 2160atacatctcg cccgtgtcac attgtggatg ggcacggtta tttcctgctt gccaggtgcc 2220cggcagggga ctccatcacc atggaattta agaaagattc cgtcagacac tcctgctcgg 2280tgccgtatga agtgaaattt aatcctgtag gcagagaact ctatactcat cccccagaac 2340acggagtaga gcaagcgtgc caagtctacg cacatgatgc acagaacaga ggagcttatg 2400tcgagatgca cctcccgggc tcagaagtgg acagcagttt ggtttccttg agcggcagtt 2460cagtcaccgt gacacctcct gatgggacta gcgccctggt ggaatgcgag tgtggctccg 2520gaggatccgg cacaaagatc tccgagacca tcaacaagac aaaacagttc agccagtgca 2580caaagaagga gcagtgcaga gcatatcggc tgcagaacga taagtgggtg tataattctg 2640acaaactgcc caaagcagcg ggagccacct taaaaggaaa actgcatgtc ccattcttgc

2700tggcagacgg caaatgcacc gtgcctctag caccagaacc tatgataacc ttcggtttca 2760gatcagtgtc actgaaactg caccctaaga atcccacata tctaatcacc cgccaacttg 2820ctgatgagcc tcactacacg cacgagctca tatctgaacc agctgttagg aattttaccg 2880tcaccgaaaa agggtgggag tttgtatggg gaaaccaccc gccgaaaagg ttttgggcac 2940aggaaacagc acccggaaat ccacatgggc taccgcacga ggtgataact cattattacc 3000acagataccc tatgtccacc atcctgggtt tgtcaatttg tgccgccatt gcaaccgttt 3060ccgttgcagc gtctacctgg ctgttttgca gatcaagagt tgcgtgccta actccttacc 3120ggctaacacc taacgctagg ataccatttt gtctggctgt gctttgctgc gcccgcactg 3180cccgggccga gaccacctgg gagtccttgg atcacctatg gaacaataac caacagatgt 3240tctggattca attgctgatc cctctggccg ccttgatcgt agtgactcgc ctgctcaggt 3300gcgtgtgctg tgtcgtgcct tttttagtca tggccggcgc cgcaggcgcc ggcgcctacg 3360agcacgcgac cacgatgccg agccaagcgg gaatctcgta taacactata gtcaacagag 3420caggctacgc accactccct atcagcataa caccaacaaa gatcaagctg atacctacag 3480tgaacttgga gtacgtcacc tgccactaca aaacaggaat ggattcacca gccatcaaat 3540gctgcggatc tcaggaatgc actccaactt acaggcctga tgaacagtgc aaagtcttca 3600caggggttta cccgttcatg tggggtggtg catattgctt ttgcgacact gagaacaccc 3660aagtcagcaa ggcctacgta atgaaatctg acgactgcct tgcggatcat gctgaagcat 3720ataaagcgca cacagcctca gtgcaggcgt tcctcaacat cacagtggga gaacactcta 3780ttgtgactac cgtgtatgtg aatggagaaa ctcctgtgaa tttcaatggg gtcaaaataa 3840ctgcaggtcc gctttccaca gcttggacac cctttgatcg caaaatcgtg cagtatgccg 3900gggagatcta taattatgat tttcctgagt atggggcagg acaaccagga gcatttggag 3960atatacaatc cagaacagtc tcaagctctg atctgtatgc caataccaac ctagtgctgc 4020agagacccaa agcaggagcg atccacgtgc catacactca ggcaccttcg ggttttgagc 4080aatggaagaa agataaagct ccatcattga aatttaccgc ccctttcgga tgcgaaatat 4140atacaaaccc cattcgcgcc gaaaactgtg ctgtagggtc aattccatta gcctttgaca 4200ttcccgacgc cttgttcacc agggtgtcag aaacaccgac actttcagcg gccgaatgca 4260ctcttaacga gtgcgtgtat tcttccgact ttggtgggat cgccacggtc aagtactcgg 4320ccagcaagtc aggcaagtgc gcagtccatg tgccatcagg gactgctacc ctaaaagaag 4380cagcagtcga gctaaccgag caagggtcgg cgactatcca tttctcgacc gcaaatatcc 4440acccggagtt caggctccaa atatgcacat catatgttac gtgcaaaggt gattgtcacc 4500ccccgaaaga ccatattgtg acacaccctc agtatcacgc ccaaacattt acagccgcgg 4560tgtcaaaaac cgcgtggacg tggttaacat ccctgctggg aggatcagcc gtaattatta 4620taattggctt ggtgctggct actattgtgg ccatgtacgt gctgaccaac cagaaacata 4680attaaggatc tagatctgct gtgccttcta gttgccagcc atctgttgtt tgcccctccc 4740ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg 4800aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg 4860acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta 4920tgggtaccca ggtgctgaag aattgacccg gttcctcctg ggccagaaag aagcaggcac 4980atccccttct ctgtgacaca ccctgtccac gcccctggtt cttagttcca gccccactca 5040taggacactc atagctcagg agggctccgc cttcaatccc acccgctaaa gtacttggag 5100cggtctctcc ctccctcatc agcccaccaa accaaaccta gcctccaaga gtgggaagaa 5160attaaagcaa gataggctat taagtgcaga gggagagaaa atgcctccaa catgtgagga 5220agtaatgaga gaaatcatag aattttaagg ccatgattta aggccatcat ggcctaagct 5280tgaaaggaga taggatcaaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 5340ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 5400gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 5460gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 5520tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 5580gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 5640ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 5700ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 5760acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 5820tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 5880ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 5940ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 6000ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 6060actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 6120gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 6180tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 6240caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 6300atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 6360acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 6420ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 6480ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 6540tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 6600tgctgcaatg ataccgcgag aaccacgctc accggctcca gatttatcag caataaacca 6660gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 6720tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 6780tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 6840ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 6900tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 6960ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 7020gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 7080ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 7140cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 7200ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 7260ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 7320gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 7380ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 7440gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt 7500aacctataaa aataggcgta tcacgaggcc ctttcgggtc gcgcgtttcg gtgatgacgg 7560tgaaaacctc tgacacatgc agctcccgtt gacggtcaca gcttgtctgt aagcggatgc 7620cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct 7680taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa 7740tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc 7800cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagggt tgagtgttgt 7860tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa 7920aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg 7980gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg 8040acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc 8100tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa 8160tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac 8220agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt 8280tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt 8340gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg 8400acggccagtg aattccatgg tctcaacttt c 8431133750DNAArtificial SequenceSynthetic vector VLP_CHI 512 -CHIKV VLP 13atggagttca tcccgacgca aactttctat aacagaaggt accaaccccg accctgggcc 60ccacgcccta caattcaagt aattagacct agaccacgtc cacagaggca ggctgggcaa 120ctcgcccagc tgatctccgc agtcaacaaa ttgaccatgc gcgcggtacc tcaacagaag 180cctcgcagaa atcggaaaaa caagaagcaa aggcagaaga agcaggcgcc gcaaaacgac 240ccaaagcaaa agaagcaacc accacaaaag aagccggctc aaaagaagaa gaaaccaggc 300cgtagggaga gaatgtgcat gaaaattgaa aatgattgca tcttcgaagt caagcatgaa 360ggcaaagtga tgggctacgc atgcctggtg ggggataaag taatgaaacc agcacatgtg 420aagggaacta tcgacaatgc cgatctggct aaactggcct ttaagcggtc gtctaaatac 480gatcttgaat gtgcacagat accggtgcac atgaagtctg atgcctcgaa gtttacccac 540gagaaacccg aggggtacta taactggcat cacggagcag tgcagtattc aggaggccgg 600ttcactatcc cgacgggtgc aggcaagccg ggagacagcg gcagaccgat cttcgacaac 660aaaggacggg tggtggccat cgtcctagga ggggccaacg aaggtgcccg cacggccctc 720tccgtggtga cgtggaacaa agacatcgtc acaaaaatta cccctgaggg agccgaagag 780tggagcctcg ccctcccggt cttgtgcctg ttggcaaaca ctacattccc ctgctctcag 840ccgccttgca caccctgctg ctacgaaaag gaaccggaaa gcaccttgcg catgcttgag 900gacaacgtga tgagacccgg atactaccag ctactaaaag catcgctgac ttgctctccc 960caccgccaaa gacgcagtac taaggacaat tttaatgtct ataaagccac aagaccatat 1020ctagctcatt gtcctgactg cggagaaggg cattcgtgcc acagccctat cgcattggag 1080cgcatcagaa atgaagcaac ggacggaacg ctgaaaatcc aggtctcttt gcagatcggg 1140ataaagacag atgacagcca cgattggacc aagctgcgct atatggatag ccatacgcca 1200gcggacgcgg agcgagccgg attgcttgta aggacttcag caccgtgcac gatcaccggg 1260accatgggac actttattct cgcccgatgc ccgaaaggag agacgctgac agtgggattt 1320acggacagca gaaagatcag ccacacatgc acacacccgt tccatcatga accacctgtg 1380ataggtaggg agaggttcca ctctcgacca caacatggta aagagttacc ttgcagcacg 1440tacgtgcaga gcaccgctgc cactgctgag gagatagagg tgcatatgcc cccagatact 1500cctgaccgca cgctgatgac gcagcagtcc ggaggatcca acgtgaagat cacagttaat 1560gggcagacgg tgcggtacaa gtgcaactgc ggtggctcaa acgagggact gacaaccaca 1620gacaaagtga tcaataactg caaaattgat cagtgccatg ctgcagtcac taatcacaag 1680aattggcaat acaactcccc tttagtcccg cgcaacgctg aactcgggga ccgtaaagga 1740aagatccaca tcccattccc attggcaaac gtgacttgca gagtgccaaa agcaagaaac 1800cctacagtaa cttacggaaa aaaccaagtc accatgctgc tgtatcctga ccatccgaca 1860ctcttgtctt accgtaacat gggacaggaa ccaaattacc acgaggagtg ggtgacacac 1920aagaaggagg ttaccttgac cgtgcctact gagggtctgg aggtcacttg gggcaacaac 1980gaaccataca agtactggcc gcagatgtct acgaacggta ctgctcatgg tcacccacat 2040gagataatct tgtactatta tgagctgtac cccactatga ctgtagtcat tgtgtcggtg 2100gcctcgttcg tgcttctgtc gatggtgggc acagcagtgg gaatgtgtgt gtgcgcacgg 2160cgcagatgca ttacaccata tgaattaaca ccaggagcca ctgttccctt cctgctcagc 2220ctgctatgct gcgtcagaac gaccaaggcg gccacatatt acgaggctgc ggcatatcta 2280tggaacgaac agcagcccct gttctggttg caggctctta tcccgctggc cgccttgatc 2340gtcctgtgca actgtctgaa actcttgcca tgctgctgta agaccctggc ttttttagcc 2400gtaatgagca tcggtgccca cactgtgagc gcgtacgaac acgtaacagt gatcccgaac 2460acggtgggag taccgtataa gactcttgtc aacagaccgg gttacagccc catggtgttg 2520gagatggagc tacaatcagt caccttggaa ccaacactgt cacttgacta catcacgtgc 2580gagtacaaaa ctgtcatccc ctccccgtac gtgaagtgct gtggtacagc agagtgcaag 2640gacaagagcc taccagacta cagctgcaag gtctttactg gagtctaccc atttatgtgg 2700ggcggcgcct actgcttttg cgacgccgaa aatacgcaat tgagcgaggc acatgtagag 2760aaatctgaat cttgcaaaac agagtttgca tcggcctaca gagcccacac cgcatcggcg 2820tcggcgaagc tccgcgtcct ttaccaagga aacaacatta ccgtagctgc ctacgctaac 2880ggtgaccatg ccgtcacagt aaaggacgcc aagtttgtcg tgggcccaat gtcctccgcc 2940tggacacctt ttgacaacaa aatcgtggtg tacaaaggcg acgtctacaa catggactac 3000ccaccttttg gcgcaggaag accaggacaa tttggtgaca ttcaaagtcg tacaccggaa 3060agtaaagacg tttatgccaa cactcagttg gtactacaga ggccagcagc aggcacggta 3120catgtaccat actctcaggc accatctggc ttcaagtatt ggctgaagga acgaggagca 3180tcgctacagc acacggcacc gttcggttgc cagattgcga caaacccggt aagagctgta 3240aattgcgctg tggggaacat accaatttcc atcgacatac cggatgcggc ctttactagg 3300gttgtcgatg caccctctgt aacggacatg tcatgcgaag taccagcctg cactcactcc 3360tccgactttg ggggcgtcgc catcatcaaa tacacagcta gcaagaaagg taaatgtgca 3420gtacattcga tgaccaacgc cgttaccatt cgagaagccg acgtagaagt agaggggaac 3480tcccagctgc aaatatcctt ctcaacagcc ctggcaagcg ccgagtttcg cgtgcaagtg 3540tgctccacac aagtacactg cgcagccgca tgccaccctc caaaggacca catagtcaat 3600tacccagcat cacacaccac ccttggggtc caggatatat ccacaacggc aatgtcttgg 3660gtgcagaaga ttacgggagg agtaggatta attgttgctg ttgctgcctt aattttaatt 3720gtggtgctat gcgtgtcgtt tagcaggcac 3750141250PRTArtificial SequenceSynthetic vector VLP_CHI 512 -CHIKV VLP 14Met Glu Phe Ile Pro Thr Gln Thr Phe Tyr Asn Arg Arg Tyr Gln Pro 1 5 10 15 Arg Pro Trp Ala Pro Arg Pro Thr Ile Gln Val Ile Arg Pro Arg Pro 20 25 30 Arg Pro Gln Arg Gln Ala Gly Gln Leu Ala Gln Leu Ile Ser Ala Val 35 40 45 Asn Lys Leu Thr Met Arg Ala Val Pro Gln Gln Lys Pro Arg Arg Asn 50 55 60 Arg Lys Asn Lys Lys Gln Arg Gln Lys Lys Gln Ala Pro Gln Asn Asp 65 70 75 80 Pro Lys Gln Lys Lys Gln Pro Pro Gln Lys Lys Pro Ala Gln Lys Lys 85 90 95 Lys Lys Pro Gly Arg Arg Glu Arg Met Cys Met Lys Ile Glu Asn Asp 100 105 110 Cys Ile Phe Glu Val Lys His Glu Gly Lys Val Met Gly Tyr Ala Cys 115 120 125 Leu Val Gly Asp Lys Val Met Lys Pro Ala His Val Lys Gly Thr Ile 130 135 140 Asp Asn Ala Asp Leu Ala Lys Leu Ala Phe Lys Arg Ser Ser Lys Tyr 145 150 155 160 Asp Leu Glu Cys Ala Gln Ile Pro Val His Met Lys Ser Asp Ala Ser 165 170 175 Lys Phe Thr His Glu Lys Pro Glu Gly Tyr Tyr Asn Trp His His Gly 180 185 190 Ala Val Gln Tyr Ser Gly Gly Arg Phe Thr Ile Pro Thr Gly Ala Gly 195 200 205 Lys Pro Gly Asp Ser Gly Arg Pro Ile Phe Asp Asn Lys Gly Arg Val 210 215 220 Val Ala Ile Val Leu Gly Gly Ala Asn Glu Gly Ala Arg Thr Ala Leu 225 230 235 240 Ser Val Val Thr Trp Asn Lys Asp Ile Val Thr Lys Ile Thr Pro Glu 245 250 255 Gly Ala Glu Glu Trp Ser Leu Ala Leu Pro Val Leu Cys Leu Leu Ala 260 265 270 Asn Thr Thr Phe Pro Cys Ser Gln Pro Pro Cys Thr Pro Cys Cys Tyr 275 280 285 Glu Lys Glu Pro Glu Ser Thr Leu Arg Met Leu Glu Asp Asn Val Met 290 295 300 Arg Pro Gly Tyr Tyr Gln Leu Leu Lys Ala Ser Leu Thr Cys Ser Pro 305 310 315 320 His Arg Gln Arg Arg Ser Thr Lys Asp Asn Phe Asn Val Tyr Lys Ala 325 330 335 Thr Arg Pro Tyr Leu Ala His Cys Pro Asp Cys Gly Glu Gly His Ser 340 345 350 Cys His Ser Pro Ile Ala Leu Glu Arg Ile Arg Asn Glu Ala Thr Asp 355 360 365 Gly Thr Leu Lys Ile Gln Val Ser Leu Gln Ile Gly Ile Lys Thr Asp 370 375 380 Asp Ser His Asp Trp Thr Lys Leu Arg Tyr Met Asp Ser His Thr Pro 385 390 395 400 Ala Asp Ala Glu Arg Ala Gly Leu Leu Val Arg Thr Ser Ala Pro Cys 405 410 415 Thr Ile Thr Gly Thr Met Gly His Phe Ile Leu Ala Arg Cys Pro Lys 420 425 430 Gly Glu Thr Leu Thr Val Gly Phe Thr Asp Ser Arg Lys Ile Ser His 435 440 445 Thr Cys Thr His Pro Phe His His Glu Pro Pro Val Ile Gly Arg Glu 450 455 460 Arg Phe His Ser Arg Pro Gln His Gly Lys Glu Leu Pro Cys Ser Thr 465 470 475 480 Tyr Val Gln Ser Thr Ala Ala Thr Ala Glu Glu Ile Glu Val His Met 485 490 495 Pro Pro Asp Thr Pro Asp Arg Thr Leu Met Thr Gln Gln Ser Gly Gly 500 505 510 Ser Asn Val Lys Ile Thr Val Asn Gly Gln Thr Val Arg Tyr Lys Cys 515 520 525 Asn Cys Gly Gly Ser Asn Glu Gly Leu Thr Thr Thr Asp Lys Val Ile 530 535 540 Asn Asn Cys Lys Ile Asp Gln Cys His Ala Ala Val Thr Asn His Lys 545 550 555 560 Asn Trp Gln Tyr Asn Ser Pro Leu Val Pro Arg Asn Ala Glu Leu Gly 565 570 575 Asp Arg Lys Gly Lys Ile His Ile Pro Phe Pro Leu Ala Asn Val Thr 580 585 590 Cys Arg Val Pro Lys Ala Arg Asn Pro Thr Val Thr Tyr Gly Lys Asn 595 600 605 Gln Val Thr Met Leu Leu Tyr Pro Asp His Pro Thr Leu Leu Ser Tyr 610 615 620 Arg Asn Met Gly Gln Glu Pro Asn Tyr His Glu Glu Trp Val Thr His 625 630 635 640 Lys Lys Glu Val Thr Leu Thr Val Pro Thr Glu Gly Leu Glu Val Thr 645 650 655 Trp Gly Asn Asn Glu Pro Tyr Lys Tyr Trp Pro Gln Met Ser Thr Asn 660 665 670 Gly Thr Ala His Gly His Pro His Glu Ile Ile Leu Tyr Tyr Tyr Glu 675 680 685 Leu Tyr Pro Thr Met Thr Val Val Ile Val Ser Val Ala Ser Phe Val 690 695 700 Leu Leu Ser Met Val Gly Thr Ala Val Gly Met Cys Val Cys Ala Arg 705 710 715 720 Arg Arg Cys Ile Thr Pro Tyr Glu Leu Thr Pro Gly Ala Thr Val Pro 725 730 735 Phe Leu Leu Ser Leu Leu Cys Cys Val Arg Thr Thr Lys Ala Ala Thr 740 745 750 Tyr Tyr Glu Ala Ala Ala Tyr Leu Trp Asn Glu Gln Gln Pro Leu Phe 755 760 765 Trp Leu Gln Ala Leu Ile Pro Leu Ala Ala Leu Ile Val Leu Cys Asn 770 775 780 Cys Leu Lys Leu Leu Pro Cys Cys Cys Lys Thr Leu Ala Phe Leu Ala 785 790 795 800 Val Met Ser Ile Gly Ala His Thr Val Ser Ala Tyr Glu His Val Thr 805 810 815 Val Ile Pro Asn Thr Val Gly Val Pro Tyr Lys Thr Leu Val Asn Arg 820

825 830 Pro Gly Tyr Ser Pro Met Val Leu Glu Met Glu Leu Gln Ser Val Thr 835 840 845 Leu Glu Pro Thr Leu Ser Leu Asp Tyr Ile Thr Cys Glu Tyr Lys Thr 850 855 860 Val Ile Pro Ser Pro Tyr Val Lys Cys Cys Gly Thr Ala Glu Cys Lys 865 870 875 880 Asp Lys Ser Leu Pro Asp Tyr Ser Cys Lys Val Phe Thr Gly Val Tyr 885 890 895 Pro Phe Met Trp Gly Gly Ala Tyr Cys Phe Cys Asp Ala Glu Asn Thr 900 905 910 Gln Leu Ser Glu Ala His Val Glu Lys Ser Glu Ser Cys Lys Thr Glu 915 920 925 Phe Ala Ser Ala Tyr Arg Ala His Thr Ala Ser Ala Ser Ala Lys Leu 930 935 940 Arg Val Leu Tyr Gln Gly Asn Asn Ile Thr Val Ala Ala Tyr Ala Asn 945 950 955 960 Gly Asp His Ala Val Thr Val Lys Asp Ala Lys Phe Val Val Gly Pro 965 970 975 Met Ser Ser Ala Trp Thr Pro Phe Asp Asn Lys Ile Val Val Tyr Lys 980 985 990 Gly Asp Val Tyr Asn Met Asp Tyr Pro Pro Phe Gly Ala Gly Arg Pro 995 1000 1005 Gly Gln Phe Gly Asp Ile Gln Ser Arg Thr Pro Glu Ser Lys Asp 1010 1015 1020 Val Tyr Ala Asn Thr Gln Leu Val Leu Gln Arg Pro Ala Ala Gly 1025 1030 1035 Thr Val His Val Pro Tyr Ser Gln Ala Pro Ser Gly Phe Lys Tyr 1040 1045 1050 Trp Leu Lys Glu Arg Gly Ala Ser Leu Gln His Thr Ala Pro Phe 1055 1060 1065 Gly Cys Gln Ile Ala Thr Asn Pro Val Arg Ala Val Asn Cys Ala 1070 1075 1080 Val Gly Asn Ile Pro Ile Ser Ile Asp Ile Pro Asp Ala Ala Phe 1085 1090 1095 Thr Arg Val Val Asp Ala Pro Ser Val Thr Asp Met Ser Cys Glu 1100 1105 1110 Val Pro Ala Cys Thr His Ser Ser Asp Phe Gly Gly Val Ala Ile 1115 1120 1125 Ile Lys Tyr Thr Ala Ser Lys Lys Gly Lys Cys Ala Val His Ser 1130 1135 1140 Met Thr Asn Ala Val Thr Ile Arg Glu Ala Asp Val Glu Val Glu 1145 1150 1155 Gly Asn Ser Gln Leu Gln Ile Ser Phe Ser Thr Ala Leu Ala Ser 1160 1165 1170 Ala Glu Phe Arg Val Gln Val Cys Ser Thr Gln Val His Cys Ala 1175 1180 1185 Ala Ala Cys His Pro Pro Lys Asp His Ile Val Asn Tyr Pro Ala 1190 1195 1200 Ser His Thr Thr Leu Gly Val Gln Asp Ile Ser Thr Thr Ala Met 1205 1210 1215 Ser Trp Val Gln Lys Ile Thr Gly Gly Val Gly Leu Ile Val Ala 1220 1225 1230 Val Ala Ala Leu Ile Leu Ile Val Val Leu Cys Val Ser Phe Ser 1235 1240 1245 Arg His 1250153756DNAArtificial SequenceSynthetic vector VLP_CHI 532 -CHIKV VLP 15atggagttca tcccgacgca aactttctat aacagaaggt accaaccccg accctgggcc 60ccacgcccta caattcaagt aattagacct agaccacgtc cacagaggca ggctgggcaa 120ctcgcccagc tgatctccgc agtcaacaaa ttgaccatgc gcgcggtacc tcaacagaag 180cctcgcagaa atcggaaaaa caagaagcaa aggcagaaga agcaggcgcc gcaaaacgac 240ccaaagcaaa agaagcaacc accacaaaag aagccggctc aaaagaagaa gaaaccaggc 300cgtagggaga gaatgtgcat gaaaattgaa aatgattgca tcttcgaagt caagcatgaa 360ggcaaagtga tgggctacgc atgcctggtg ggggataaag taatgaaacc agcacatgtg 420aagggaacta tcgacaatgc cgatctggct aaactggcct ttaagcggtc gtctaaatac 480gatcttgaat gtgcacagat accggtgcac atgaagtctg atgcctcgaa gtttacccac 540gagaaacccg aggggtacta taactggcat cacggagcag tgcagtattc aggaggccgg 600ttcactatcc cgacgggtgc aggcaagccg ggagacagcg gcagaccgat cttcgacaac 660aaaggacggg tggtggccat cgtcctagga ggggccaacg aaggtgcccg cacggccctc 720tccgtggtga cgtggaacaa agacatcgtc acaaaaatta cccctgaggg agccgaagag 780tggagcctcg ccctcccggt cttgtgcctg ttggcaaaca ctacattccc ctgctctcag 840ccgccttgca caccctgctg ctacgaaaag gaaccggaaa gcaccttgcg catgcttgag 900gacaacgtga tgagacccgg atactaccag ctactaaaag catcgctgac ttgctctccc 960caccgccaaa gacgcagtac taaggacaat tttaatgtct ataaagccac aagaccatat 1020ctagctcatt gtcctgactg cggagaaggg cattcgtgcc acagccctat cgcattggag 1080cgcatcagaa atgaagcaac ggacggaacg ctgaaaatcc aggtctcttt gcagatcggg 1140ataaagacag atgacagcca cgattggacc aagctgcgct atatggatag ccatacgcca 1200gcggacgcgg agcgagccgg attgcttgta aggacttcag caccgtgcac gatcaccggg 1260accatgggac actttattct cgcccgatgc ccgaaaggag agacgctgac agtgggattt 1320acggacagca gaaagatcag ccacacatgc acacacccgt tccatcatga accacctgtg 1380ataggtaggg agaggttcca ctctcgacca caacatggta aagagttacc ttgcagcacg 1440tacgtgcaga gcaccgctgc cactgctgag gagatagagg tgcatatgcc cccagatact 1500cctgaccgca cgctgatgac gcagcagtct ggcaacgtga agatcacagt taatgggcag 1560acggtgcggt acaagtgcaa ctgcggtggc tccggaagtg gatccaacga gggactgaca 1620accacagaca aagtgatcaa taactgcaaa attgatcagt gccatgctgc agtcactaat 1680cacaagaatt ggcaatacaa ctccccttta gtcccgcgca acgctgaact cggggaccgt 1740aaaggaaaga tccacatccc attcccattg gcaaacgtga cttgcagagt gccaaaagca 1800agaaacccta cagtaactta cggaaaaaac caagtcacca tgctgctgta tcctgaccat 1860ccgacactct tgtcttaccg taacatggga caggaaccaa attaccacga ggagtgggtg 1920acacacaaga aggaggttac cttgaccgtg cctactgagg gtctggaggt cacttggggc 1980aacaacgaac catacaagta ctggccgcag atgtctacga acggtactgc tcatggtcac 2040ccacatgaga taatcttgta ctattatgag ctgtacccca ctatgactgt agtcattgtg 2100tcggtggcct cgttcgtgct tctgtcgatg gtgggcacag cagtgggaat gtgtgtgtgc 2160gcacggcgca gatgcattac accatatgaa ttaacaccag gagccactgt tcccttcctg 2220ctcagcctgc tatgctgcgt cagaacgacc aaggcggcca catattacga ggctgcggca 2280tatctatgga acgaacagca gcccctgttc tggttgcagg ctcttatccc gctggccgcc 2340ttgatcgtcc tgtgcaactg tctgaaactc ttgccatgct gctgtaagac cctggctttt 2400ttagccgtaa tgagcatcgg tgcccacact gtgagcgcgt acgaacacgt aacagtgatc 2460ccgaacacgg tgggagtacc gtataagact cttgtcaaca gaccgggtta cagccccatg 2520gtgttggaga tggagctaca atcagtcacc ttggaaccaa cactgtcact tgactacatc 2580acgtgcgagt acaaaactgt catcccctcc ccgtacgtga agtgctgtgg tacagcagag 2640tgcaaggaca agagcctacc agactacagc tgcaaggtct ttactggagt ctacccattt 2700atgtggggcg gcgcctactg cttttgcgac gccgaaaata cgcaattgag cgaggcacat 2760gtagagaaat ctgaatcttg caaaacagag tttgcatcgg cctacagagc ccacaccgca 2820tcggcgtcgg cgaagctccg cgtcctttac caaggaaaca acattaccgt agctgcctac 2880gctaacggtg accatgccgt cacagtaaag gacgccaagt ttgtcgtggg cccaatgtcc 2940tccgcctgga caccttttga caacaaaatc gtggtgtaca aaggcgacgt ctacaacatg 3000gactacccac cttttggcgc aggaagacca ggacaatttg gtgacattca aagtcgtaca 3060ccggaaagta aagacgttta tgccaacact cagttggtac tacagaggcc agcagcaggc 3120acggtacatg taccatactc tcaggcacca tctggcttca agtattggct gaaggaacga 3180ggagcatcgc tacagcacac ggcaccgttc ggttgccaga ttgcgacaaa cccggtaaga 3240gctgtaaatt gcgctgtggg gaacatacca atttccatcg acataccgga tgcggccttt 3300actagggttg tcgatgcacc ctctgtaacg gacatgtcat gcgaagtacc agcctgcact 3360cactcctccg actttggggg cgtcgccatc atcaaataca cagctagcaa gaaaggtaaa 3420tgtgcagtac attcgatgac caacgccgtt accattcgag aagccgacgt agaagtagag 3480gggaactccc agctgcaaat atccttctca acagccctgg caagcgccga gtttcgcgtg 3540caagtgtgct ccacacaagt acactgcgca gccgcatgcc accctccaaa ggaccacata 3600gtcaattacc cagcatcaca caccaccctt ggggtccagg atatatccac aacggcaatg 3660tcttgggtgc agaagattac gggaggagta ggattaattg ttgctgttgc tgccttaatt 3720ttaattgtgg tgctatgcgt gtcgtttagc aggcac 3756161252PRTArtificial SequenceSynthetic vector VLP_CHI 532 -CHIKV VLP 16Met Glu Phe Ile Pro Thr Gln Thr Phe Tyr Asn Arg Arg Tyr Gln Pro 1 5 10 15 Arg Pro Trp Ala Pro Arg Pro Thr Ile Gln Val Ile Arg Pro Arg Pro 20 25 30 Arg Pro Gln Arg Gln Ala Gly Gln Leu Ala Gln Leu Ile Ser Ala Val 35 40 45 Asn Lys Leu Thr Met Arg Ala Val Pro Gln Gln Lys Pro Arg Arg Asn 50 55 60 Arg Lys Asn Lys Lys Gln Arg Gln Lys Lys Gln Ala Pro Gln Asn Asp 65 70 75 80 Pro Lys Gln Lys Lys Gln Pro Pro Gln Lys Lys Pro Ala Gln Lys Lys 85 90 95 Lys Lys Pro Gly Arg Arg Glu Arg Met Cys Met Lys Ile Glu Asn Asp 100 105 110 Cys Ile Phe Glu Val Lys His Glu Gly Lys Val Met Gly Tyr Ala Cys 115 120 125 Leu Val Gly Asp Lys Val Met Lys Pro Ala His Val Lys Gly Thr Ile 130 135 140 Asp Asn Ala Asp Leu Ala Lys Leu Ala Phe Lys Arg Ser Ser Lys Tyr 145 150 155 160 Asp Leu Glu Cys Ala Gln Ile Pro Val His Met Lys Ser Asp Ala Ser 165 170 175 Lys Phe Thr His Glu Lys Pro Glu Gly Tyr Tyr Asn Trp His His Gly 180 185 190 Ala Val Gln Tyr Ser Gly Gly Arg Phe Thr Ile Pro Thr Gly Ala Gly 195 200 205 Lys Pro Gly Asp Ser Gly Arg Pro Ile Phe Asp Asn Lys Gly Arg Val 210 215 220 Val Ala Ile Val Leu Gly Gly Ala Asn Glu Gly Ala Arg Thr Ala Leu 225 230 235 240 Ser Val Val Thr Trp Asn Lys Asp Ile Val Thr Lys Ile Thr Pro Glu 245 250 255 Gly Ala Glu Glu Trp Ser Leu Ala Leu Pro Val Leu Cys Leu Leu Ala 260 265 270 Asn Thr Thr Phe Pro Cys Ser Gln Pro Pro Cys Thr Pro Cys Cys Tyr 275 280 285 Glu Lys Glu Pro Glu Ser Thr Leu Arg Met Leu Glu Asp Asn Val Met 290 295 300 Arg Pro Gly Tyr Tyr Gln Leu Leu Lys Ala Ser Leu Thr Cys Ser Pro 305 310 315 320 His Arg Gln Arg Arg Ser Thr Lys Asp Asn Phe Asn Val Tyr Lys Ala 325 330 335 Thr Arg Pro Tyr Leu Ala His Cys Pro Asp Cys Gly Glu Gly His Ser 340 345 350 Cys His Ser Pro Ile Ala Leu Glu Arg Ile Arg Asn Glu Ala Thr Asp 355 360 365 Gly Thr Leu Lys Ile Gln Val Ser Leu Gln Ile Gly Ile Lys Thr Asp 370 375 380 Asp Ser His Asp Trp Thr Lys Leu Arg Tyr Met Asp Ser His Thr Pro 385 390 395 400 Ala Asp Ala Glu Arg Ala Gly Leu Leu Val Arg Thr Ser Ala Pro Cys 405 410 415 Thr Ile Thr Gly Thr Met Gly His Phe Ile Leu Ala Arg Cys Pro Lys 420 425 430 Gly Glu Thr Leu Thr Val Gly Phe Thr Asp Ser Arg Lys Ile Ser His 435 440 445 Thr Cys Thr His Pro Phe His His Glu Pro Pro Val Ile Gly Arg Glu 450 455 460 Arg Phe His Ser Arg Pro Gln His Gly Lys Glu Leu Pro Cys Ser Thr 465 470 475 480 Tyr Val Gln Ser Thr Ala Ala Thr Ala Glu Glu Ile Glu Val His Met 485 490 495 Pro Pro Asp Thr Pro Asp Arg Thr Leu Met Thr Gln Gln Ser Gly Asn 500 505 510 Val Lys Ile Thr Val Asn Gly Gln Thr Val Arg Tyr Lys Cys Asn Cys 515 520 525 Gly Gly Ser Gly Ser Gly Ser Asn Glu Gly Leu Thr Thr Thr Asp Lys 530 535 540 Val Ile Asn Asn Cys Lys Ile Asp Gln Cys His Ala Ala Val Thr Asn 545 550 555 560 His Lys Asn Trp Gln Tyr Asn Ser Pro Leu Val Pro Arg Asn Ala Glu 565 570 575 Leu Gly Asp Arg Lys Gly Lys Ile His Ile Pro Phe Pro Leu Ala Asn 580 585 590 Val Thr Cys Arg Val Pro Lys Ala Arg Asn Pro Thr Val Thr Tyr Gly 595 600 605 Lys Asn Gln Val Thr Met Leu Leu Tyr Pro Asp His Pro Thr Leu Leu 610 615 620 Ser Tyr Arg Asn Met Gly Gln Glu Pro Asn Tyr His Glu Glu Trp Val 625 630 635 640 Thr His Lys Lys Glu Val Thr Leu Thr Val Pro Thr Glu Gly Leu Glu 645 650 655 Val Thr Trp Gly Asn Asn Glu Pro Tyr Lys Tyr Trp Pro Gln Met Ser 660 665 670 Thr Asn Gly Thr Ala His Gly His Pro His Glu Ile Ile Leu Tyr Tyr 675 680 685 Tyr Glu Leu Tyr Pro Thr Met Thr Val Val Ile Val Ser Val Ala Ser 690 695 700 Phe Val Leu Leu Ser Met Val Gly Thr Ala Val Gly Met Cys Val Cys 705 710 715 720 Ala Arg Arg Arg Cys Ile Thr Pro Tyr Glu Leu Thr Pro Gly Ala Thr 725 730 735 Val Pro Phe Leu Leu Ser Leu Leu Cys Cys Val Arg Thr Thr Lys Ala 740 745 750 Ala Thr Tyr Tyr Glu Ala Ala Ala Tyr Leu Trp Asn Glu Gln Gln Pro 755 760 765 Leu Phe Trp Leu Gln Ala Leu Ile Pro Leu Ala Ala Leu Ile Val Leu 770 775 780 Cys Asn Cys Leu Lys Leu Leu Pro Cys Cys Cys Lys Thr Leu Ala Phe 785 790 795 800 Leu Ala Val Met Ser Ile Gly Ala His Thr Val Ser Ala Tyr Glu His 805 810 815 Val Thr Val Ile Pro Asn Thr Val Gly Val Pro Tyr Lys Thr Leu Val 820 825 830 Asn Arg Pro Gly Tyr Ser Pro Met Val Leu Glu Met Glu Leu Gln Ser 835 840 845 Val Thr Leu Glu Pro Thr Leu Ser Leu Asp Tyr Ile Thr Cys Glu Tyr 850 855 860 Lys Thr Val Ile Pro Ser Pro Tyr Val Lys Cys Cys Gly Thr Ala Glu 865 870 875 880 Cys Lys Asp Lys Ser Leu Pro Asp Tyr Ser Cys Lys Val Phe Thr Gly 885 890 895 Val Tyr Pro Phe Met Trp Gly Gly Ala Tyr Cys Phe Cys Asp Ala Glu 900 905 910 Asn Thr Gln Leu Ser Glu Ala His Val Glu Lys Ser Glu Ser Cys Lys 915 920 925 Thr Glu Phe Ala Ser Ala Tyr Arg Ala His Thr Ala Ser Ala Ser Ala 930 935 940 Lys Leu Arg Val Leu Tyr Gln Gly Asn Asn Ile Thr Val Ala Ala Tyr 945 950 955 960 Ala Asn Gly Asp His Ala Val Thr Val Lys Asp Ala Lys Phe Val Val 965 970 975 Gly Pro Met Ser Ser Ala Trp Thr Pro Phe Asp Asn Lys Ile Val Val 980 985 990 Tyr Lys Gly Asp Val Tyr Asn Met Asp Tyr Pro Pro Phe Gly Ala Gly 995 1000 1005 Arg Pro Gly Gln Phe Gly Asp Ile Gln Ser Arg Thr Pro Glu Ser 1010 1015 1020 Lys Asp Val Tyr Ala Asn Thr Gln Leu Val Leu Gln Arg Pro Ala 1025 1030 1035 Ala Gly Thr Val His Val Pro Tyr Ser Gln Ala Pro Ser Gly Phe 1040 1045 1050 Lys Tyr Trp Leu Lys Glu Arg Gly Ala Ser Leu Gln His Thr Ala 1055 1060 1065 Pro Phe Gly Cys Gln Ile Ala Thr Asn Pro Val Arg Ala Val Asn 1070 1075 1080 Cys Ala Val Gly Asn Ile Pro Ile Ser Ile Asp Ile Pro Asp Ala 1085 1090 1095 Ala Phe Thr Arg Val Val Asp Ala Pro Ser Val Thr Asp Met Ser 1100 1105 1110 Cys Glu Val Pro Ala Cys Thr His Ser Ser Asp Phe Gly Gly Val 1115 1120 1125 Ala Ile Ile Lys Tyr Thr Ala Ser Lys Lys Gly Lys Cys Ala Val 1130 1135 1140 His Ser Met Thr Asn Ala Val Thr Ile Arg Glu Ala Asp Val Glu 1145 1150 1155 Val Glu Gly Asn Ser Gln Leu Gln Ile Ser Phe Ser Thr Ala Leu 1160 1165 1170 Ala Ser Ala Glu Phe Arg Val Gln Val Cys Ser Thr Gln Val His 1175 1180 1185 Cys Ala Ala Ala Cys His Pro Pro Lys Asp His Ile Val Asn Tyr 1190 1195 1200 Pro Ala Ser His Thr Thr Leu Gly Val Gln Asp Ile Ser Thr Thr 1205 1210 1215 Ala Met Ser Trp Val Gln Lys Ile Thr Gly Gly Val Gly Leu Ile 1220 1225 1230 Val Ala Val Ala Ala Leu Ile Leu Ile Val Val Leu Cys Val Ser 1235 1240 1245 Phe Ser Arg His 1250 173777DNAArtificial SequenceSynthetic vector

VLP_VEEV VLP 518 -VEEV VLP 17atgttcccgt tccagccaat gtatccgatg cagccaatgc cctatcgcaa cccgttcgcg 60gccccgcgca ggccctggtt ccccagaacc gacccttttc tggcgatgca ggtgcaggaa 120ttaacccgct cgatggctaa cctgacgttc aagcaacgcc gggacgcgcc acctgagggg 180ccatccgcta ataaaccgaa gaaggaggcc tcgcaaaaac agaaaggggg aggccaaggg 240aagaagaaga agaaccaagg gaagaagaag gctaagacag ggccgcctaa tccgaaggca 300cagaatggaa acaagaagaa gaccaacaag aaaccaggca agagacagcg catggtcatg 360aaattggaat ctgacaagac gttcccaatc atgttggaag ggaagataaa cggctacgct 420tgtgtggtcg gagggaagtt attcaggccg atgcatgtgg aaggcaagat cgacaacgac 480gttctggccg cgcttaagac gaagaaagca tccaaatacg atcttgagta tgcagatgtg 540ccacagaaca tgcgggccga tacattcaaa tacacccatg agaaacccca aggctattac 600agctggcatc atggagcagt ccaatatgaa aatgggcgtt tcacggtgcc gaaaggagtt 660ggggccaagg gagacagcgg acgacccatt ctggataacc agggacgggt ggtcgctatt 720gtgctgggag gtgtgaatga aggatctagg acagcccttt cagtcgtcat gtggaacgag 780aagggagtta ccgtgaagta tactccagag aactgcgagc aatggtcact agtgaccacc 840atgtgtctgc tcgccaatgt gacgttccca tgtgctcaac caccaatttg ctacgacaga 900aaaccagcag agactttggc catgctcagc gttaacgttg acaacccggg ctacgatgag 960ctgctggaag cagctgttaa gtgccccgga aggaaaagga gatccaccga ggagctgttt 1020aatgagtata agctaacgcg cccttacatg gccagatgca tcagatgtgc agttgggagc 1080tgccatagtc caatagcaat cgaggcagta aagagcgacg ggcacgacgg ttatgttaga 1140cttcagactt cctcgcagta tggcctggat tcctccggca acttaaaggg caggaccatg 1200cggtatgaca tgcacgggac cattaaagag ataccactac atcaagtgtc actctataca 1260tctcgcccgt gtcacattgt ggatgggcac ggttatttcc tgcttgccag gtgcccggca 1320ggggactcca tcaccatgga atttaagaaa gattccgtca gacactcctg ctcggtgccg 1380tatgaagtga aatttaatcc tgtaggcaga gaactctata ctcatccccc agaacacgga 1440gtagagcaag cgtgccaagt ctacgcacat gatgcacaga acagaggagc ttatgtcgag 1500atgcacctcc cgggctcaga agtggacagc agtttggttt ccttgagcgg ctccggagga 1560tccagttcag tcaccgtgac acctcctgat gggactagcg ccctggtgga atgcgagtgt 1620ggcggcacaa agatctccga gaccatcaac aagacaaaac agttcagcca gtgcacaaag 1680aaggagcagt gcagagcata tcggctgcag aacgataagt gggtgtataa ttctgacaaa 1740ctgcccaaag cagcgggagc caccttaaaa ggaaaactgc atgtcccatt cttgctggca 1800gacggcaaat gcaccgtgcc tctagcacca gaacctatga taaccttcgg tttcagatca 1860gtgtcactga aactgcaccc taagaatccc acatatctaa tcacccgcca acttgctgat 1920gagcctcact acacgcacga gctcatatct gaaccagctg ttaggaattt taccgtcacc 1980gaaaaagggt gggagtttgt atggggaaac cacccgccga aaaggttttg ggcacaggaa 2040acagcacccg gaaatccaca tgggctaccg cacgaggtga taactcatta ttaccacaga 2100taccctatgt ccaccatcct gggtttgtca atttgtgccg ccattgcaac cgtttccgtt 2160gcagcgtcta cctggctgtt ttgcagatca agagttgcgt gcctaactcc ttaccggcta 2220acacctaacg ctaggatacc attttgtctg gctgtgcttt gctgcgcccg cactgcccgg 2280gccgagacca cctgggagtc cttggatcac ctatggaaca ataaccaaca gatgttctgg 2340attcaattgc tgatccctct ggccgccttg atcgtagtga ctcgcctgct caggtgcgtg 2400tgctgtgtcg tgcctttttt agtcatggcc ggcgccgcag gcgccggcgc ctacgagcac 2460gcgaccacga tgccgagcca agcgggaatc tcgtataaca ctatagtcaa cagagcaggc 2520tacgcaccac tccctatcag cataacacca acaaagatca agctgatacc tacagtgaac 2580ttggagtacg tcacctgcca ctacaaaaca ggaatggatt caccagccat caaatgctgc 2640ggatctcagg aatgcactcc aacttacagg cctgatgaac agtgcaaagt cttcacaggg 2700gtttacccgt tcatgtgggg tggtgcatat tgcttttgcg acactgagaa cacccaagtc 2760agcaaggcct acgtaatgaa atctgacgac tgccttgcgg atcatgctga agcatataaa 2820gcgcacacag cctcagtgca ggcgttcctc aacatcacag tgggagaaca ctctattgtg 2880actaccgtgt atgtgaatgg agaaactcct gtgaatttca atggggtcaa aataactgca 2940ggtccgcttt ccacagcttg gacacccttt gatcgcaaaa tcgtgcagta tgccggggag 3000atctataatt atgattttcc tgagtatggg gcaggacaac caggagcatt tggagatata 3060caatccagaa cagtctcaag ctctgatctg tatgccaata ccaacctagt gctgcagaga 3120cccaaagcag gagcgatcca cgtgccatac actcaggcac cttcgggttt tgagcaatgg 3180aagaaagata aagctccatc attgaaattt accgcccctt tcggatgcga aatatataca 3240aaccccattc gcgccgaaaa ctgtgctgta gggtcaattc cattagcctt tgacattccc 3300gacgccttgt tcaccagggt gtcagaaaca ccgacacttt cagcggccga atgcactctt 3360aacgagtgcg tgtattcttc cgactttggt gggatcgcca cggtcaagta ctcggccagc 3420aagtcaggca agtgcgcagt ccatgtgcca tcagggactg ctaccctaaa agaagcagca 3480gtcgagctaa ccgagcaagg gtcggcgact atccatttct cgaccgcaaa tatccacccg 3540gagttcaggc tccaaatatg cacatcatat gttacgtgca aaggtgattg tcaccccccg 3600aaagaccata ttgtgacaca ccctcagtat cacgcccaaa catttacagc cgcggtgtca 3660aaaaccgcgt ggacgtggtt aacatccctg ctgggaggat cagccgtaat tattataatt 3720ggcttggtgc tggctactat tgtggccatg tacgtgctga ccaaccagaa acataat 3777181259PRTArtificial SequenceSynthetic vector VLP_VEEV VLP 518 -VEEV VLP 18Met Phe Pro Phe Gln Pro Met Tyr Pro Met Gln Pro Met Pro Tyr Arg 1 5 10 15 Asn Pro Phe Ala Ala Pro Arg Arg Pro Trp Phe Pro Arg Thr Asp Pro 20 25 30 Phe Leu Ala Met Gln Val Gln Glu Leu Thr Arg Ser Met Ala Asn Leu 35 40 45 Thr Phe Lys Gln Arg Arg Asp Ala Pro Pro Glu Gly Pro Ser Ala Asn 50 55 60 Lys Pro Lys Lys Glu Ala Ser Gln Lys Gln Lys Gly Gly Gly Gln Gly 65 70 75 80 Lys Lys Lys Lys Asn Gln Gly Lys Lys Lys Ala Lys Thr Gly Pro Pro 85 90 95 Asn Pro Lys Ala Gln Asn Gly Asn Lys Lys Lys Thr Asn Lys Lys Pro 100 105 110 Gly Lys Arg Gln Arg Met Val Met Lys Leu Glu Ser Asp Lys Thr Phe 115 120 125 Pro Ile Met Leu Glu Gly Lys Ile Asn Gly Tyr Ala Cys Val Val Gly 130 135 140 Gly Lys Leu Phe Arg Pro Met His Val Glu Gly Lys Ile Asp Asn Asp 145 150 155 160 Val Leu Ala Ala Leu Lys Thr Lys Lys Ala Ser Lys Tyr Asp Leu Glu 165 170 175 Tyr Ala Asp Val Pro Gln Asn Met Arg Ala Asp Thr Phe Lys Tyr Thr 180 185 190 His Glu Lys Pro Gln Gly Tyr Tyr Ser Trp His His Gly Ala Val Gln 195 200 205 Tyr Glu Asn Gly Arg Phe Thr Val Pro Lys Gly Val Gly Ala Lys Gly 210 215 220 Asp Ser Gly Arg Pro Ile Leu Asp Asn Gln Gly Arg Val Val Ala Ile 225 230 235 240 Val Leu Gly Gly Val Asn Glu Gly Ser Arg Thr Ala Leu Ser Val Val 245 250 255 Met Trp Asn Glu Lys Gly Val Thr Val Lys Tyr Thr Pro Glu Asn Cys 260 265 270 Glu Gln Trp Ser Leu Val Thr Thr Met Cys Leu Leu Ala Asn Val Thr 275 280 285 Phe Pro Cys Ala Gln Pro Pro Ile Cys Tyr Asp Arg Lys Pro Ala Glu 290 295 300 Thr Leu Ala Met Leu Ser Val Asn Val Asp Asn Pro Gly Tyr Asp Glu 305 310 315 320 Leu Leu Glu Ala Ala Val Lys Cys Pro Gly Arg Lys Arg Arg Ser Thr 325 330 335 Glu Glu Leu Phe Asn Glu Tyr Lys Leu Thr Arg Pro Tyr Met Ala Arg 340 345 350 Cys Ile Arg Cys Ala Val Gly Ser Cys His Ser Pro Ile Ala Ile Glu 355 360 365 Ala Val Lys Ser Asp Gly His Asp Gly Tyr Val Arg Leu Gln Thr Ser 370 375 380 Ser Gln Tyr Gly Leu Asp Ser Ser Gly Asn Leu Lys Gly Arg Thr Met 385 390 395 400 Arg Tyr Asp Met His Gly Thr Ile Lys Glu Ile Pro Leu His Gln Val 405 410 415 Ser Leu Tyr Thr Ser Arg Pro Cys His Ile Val Asp Gly His Gly Tyr 420 425 430 Phe Leu Leu Ala Arg Cys Pro Ala Gly Asp Ser Ile Thr Met Glu Phe 435 440 445 Lys Lys Asp Ser Val Arg His Ser Cys Ser Val Pro Tyr Glu Val Lys 450 455 460 Phe Asn Pro Val Gly Arg Glu Leu Tyr Thr His Pro Pro Glu His Gly 465 470 475 480 Val Glu Gln Ala Cys Gln Val Tyr Ala His Asp Ala Gln Asn Arg Gly 485 490 495 Ala Tyr Val Glu Met His Leu Pro Gly Ser Glu Val Asp Ser Ser Leu 500 505 510 Val Ser Leu Ser Gly Ser Gly Gly Ser Ser Ser Val Thr Val Thr Pro 515 520 525 Pro Asp Gly Thr Ser Ala Leu Val Glu Cys Glu Cys Gly Gly Thr Lys 530 535 540 Ile Ser Glu Thr Ile Asn Lys Thr Lys Gln Phe Ser Gln Cys Thr Lys 545 550 555 560 Lys Glu Gln Cys Arg Ala Tyr Arg Leu Gln Asn Asp Lys Trp Val Tyr 565 570 575 Asn Ser Asp Lys Leu Pro Lys Ala Ala Gly Ala Thr Leu Lys Gly Lys 580 585 590 Leu His Val Pro Phe Leu Leu Ala Asp Gly Lys Cys Thr Val Pro Leu 595 600 605 Ala Pro Glu Pro Met Ile Thr Phe Gly Phe Arg Ser Val Ser Leu Lys 610 615 620 Leu His Pro Lys Asn Pro Thr Tyr Leu Ile Thr Arg Gln Leu Ala Asp 625 630 635 640 Glu Pro His Tyr Thr His Glu Leu Ile Ser Glu Pro Ala Val Arg Asn 645 650 655 Phe Thr Val Thr Glu Lys Gly Trp Glu Phe Val Trp Gly Asn His Pro 660 665 670 Pro Lys Arg Phe Trp Ala Gln Glu Thr Ala Pro Gly Asn Pro His Gly 675 680 685 Leu Pro His Glu Val Ile Thr His Tyr Tyr His Arg Tyr Pro Met Ser 690 695 700 Thr Ile Leu Gly Leu Ser Ile Cys Ala Ala Ile Ala Thr Val Ser Val 705 710 715 720 Ala Ala Ser Thr Trp Leu Phe Cys Arg Ser Arg Val Ala Cys Leu Thr 725 730 735 Pro Tyr Arg Leu Thr Pro Asn Ala Arg Ile Pro Phe Cys Leu Ala Val 740 745 750 Leu Cys Cys Ala Arg Thr Ala Arg Ala Glu Thr Thr Trp Glu Ser Leu 755 760 765 Asp His Leu Trp Asn Asn Asn Gln Gln Met Phe Trp Ile Gln Leu Leu 770 775 780 Ile Pro Leu Ala Ala Leu Ile Val Val Thr Arg Leu Leu Arg Cys Val 785 790 795 800 Cys Cys Val Val Pro Phe Leu Val Met Ala Gly Ala Ala Gly Ala Gly 805 810 815 Ala Tyr Glu His Ala Thr Thr Met Pro Ser Gln Ala Gly Ile Ser Tyr 820 825 830 Asn Thr Ile Val Asn Arg Ala Gly Tyr Ala Pro Leu Pro Ile Ser Ile 835 840 845 Thr Pro Thr Lys Ile Lys Leu Ile Pro Thr Val Asn Leu Glu Tyr Val 850 855 860 Thr Cys His Tyr Lys Thr Gly Met Asp Ser Pro Ala Ile Lys Cys Cys 865 870 875 880 Gly Ser Gln Glu Cys Thr Pro Thr Tyr Arg Pro Asp Glu Gln Cys Lys 885 890 895 Val Phe Thr Gly Val Tyr Pro Phe Met Trp Gly Gly Ala Tyr Cys Phe 900 905 910 Cys Asp Thr Glu Asn Thr Gln Val Ser Lys Ala Tyr Val Met Lys Ser 915 920 925 Asp Asp Cys Leu Ala Asp His Ala Glu Ala Tyr Lys Ala His Thr Ala 930 935 940 Ser Val Gln Ala Phe Leu Asn Ile Thr Val Gly Glu His Ser Ile Val 945 950 955 960 Thr Thr Val Tyr Val Asn Gly Glu Thr Pro Val Asn Phe Asn Gly Val 965 970 975 Lys Ile Thr Ala Gly Pro Leu Ser Thr Ala Trp Thr Pro Phe Asp Arg 980 985 990 Lys Ile Val Gln Tyr Ala Gly Glu Ile Tyr Asn Tyr Asp Phe Pro Glu 995 1000 1005 Tyr Gly Ala Gly Gln Pro Gly Ala Phe Gly Asp Ile Gln Ser Arg 1010 1015 1020 Thr Val Ser Ser Ser Asp Leu Tyr Ala Asn Thr Asn Leu Val Leu 1025 1030 1035 Gln Arg Pro Lys Ala Gly Ala Ile His Val Pro Tyr Thr Gln Ala 1040 1045 1050 Pro Ser Gly Phe Glu Gln Trp Lys Lys Asp Lys Ala Pro Ser Leu 1055 1060 1065 Lys Phe Thr Ala Pro Phe Gly Cys Glu Ile Tyr Thr Asn Pro Ile 1070 1075 1080 Arg Ala Glu Asn Cys Ala Val Gly Ser Ile Pro Leu Ala Phe Asp 1085 1090 1095 Ile Pro Asp Ala Leu Phe Thr Arg Val Ser Glu Thr Pro Thr Leu 1100 1105 1110 Ser Ala Ala Glu Cys Thr Leu Asn Glu Cys Val Tyr Ser Ser Asp 1115 1120 1125 Phe Gly Gly Ile Ala Thr Val Lys Tyr Ser Ala Ser Lys Ser Gly 1130 1135 1140 Lys Cys Ala Val His Val Pro Ser Gly Thr Ala Thr Leu Lys Glu 1145 1150 1155 Ala Ala Val Glu Leu Thr Glu Gln Gly Ser Ala Thr Ile His Phe 1160 1165 1170 Ser Thr Ala Asn Ile His Pro Glu Phe Arg Leu Gln Ile Cys Thr 1175 1180 1185 Ser Tyr Val Thr Cys Lys Gly Asp Cys His Pro Pro Lys Asp His 1190 1195 1200 Ile Val Thr His Pro Gln Tyr His Ala Gln Thr Phe Thr Ala Ala 1205 1210 1215 Val Ser Lys Thr Ala Trp Thr Trp Leu Thr Ser Leu Leu Gly Gly 1220 1225 1230 Ser Ala Val Ile Ile Ile Ile Gly Leu Val Leu Ala Thr Ile Val 1235 1240 1245 Ala Met Tyr Val Leu Thr Asn Gln Lys His Asn 1250 1255 193777DNAArtificial SequenceSynthetic vector VLP_VEEV VLP 519 -VEEV VLP 19atgttcccgt tccagccaat gtatccgatg cagccaatgc cctatcgcaa cccgttcgcg 60gccccgcgca ggccctggtt ccccagaacc gacccttttc tggcgatgca ggtgcaggaa 120ttaacccgct cgatggctaa cctgacgttc aagcaacgcc gggacgcgcc acctgagggg 180ccatccgcta ataaaccgaa gaaggaggcc tcgcaaaaac agaaaggggg aggccaaggg 240aagaagaaga agaaccaagg gaagaagaag gctaagacag ggccgcctaa tccgaaggca 300cagaatggaa acaagaagaa gaccaacaag aaaccaggca agagacagcg catggtcatg 360aaattggaat ctgacaagac gttcccaatc atgttggaag ggaagataaa cggctacgct 420tgtgtggtcg gagggaagtt attcaggccg atgcatgtgg aaggcaagat cgacaacgac 480gttctggccg cgcttaagac gaagaaagca tccaaatacg atcttgagta tgcagatgtg 540ccacagaaca tgcgggccga tacattcaaa tacacccatg agaaacccca aggctattac 600agctggcatc atggagcagt ccaatatgaa aatgggcgtt tcacggtgcc gaaaggagtt 660ggggccaagg gagacagcgg acgacccatt ctggataacc agggacgggt ggtcgctatt 720gtgctgggag gtgtgaatga aggatctagg acagcccttt cagtcgtcat gtggaacgag 780aagggagtta ccgtgaagta tactccagag aactgcgagc aatggtcact agtgaccacc 840atgtgtctgc tcgccaatgt gacgttccca tgtgctcaac caccaatttg ctacgacaga 900aaaccagcag agactttggc catgctcagc gttaacgttg acaacccggg ctacgatgag 960ctgctggaag cagctgttaa gtgccccgga aggaaaagga gatccaccga ggagctgttt 1020aatgagtata agctaacgcg cccttacatg gccagatgca tcagatgtgc agttgggagc 1080tgccatagtc caatagcaat cgaggcagta aagagcgacg ggcacgacgg ttatgttaga 1140cttcagactt cctcgcagta tggcctggat tcctccggca acttaaaggg caggaccatg 1200cggtatgaca tgcacgggac cattaaagag ataccactac atcaagtgtc actctataca 1260tctcgcccgt gtcacattgt ggatgggcac ggttatttcc tgcttgccag gtgcccggca 1320ggggactcca tcaccatgga atttaagaaa gattccgtca gacactcctg ctcggtgccg 1380tatgaagtga aatttaatcc tgtaggcaga gaactctata ctcatccccc agaacacgga 1440gtagagcaag cgtgccaagt ctacgcacat gatgcacaga acagaggagc ttatgtcgag 1500atgcacctcc cgggctcaga agtggacagc agtttggttt ccttgagcgg cagttccgga 1560ggatcctcag tcaccgtgac acctcctgat gggactagcg ccctggtgga atgcgagtgt 1620ggcggcacaa agatctccga gaccatcaac aagacaaaac agttcagcca gtgcacaaag 1680aaggagcagt gcagagcata tcggctgcag aacgataagt gggtgtataa ttctgacaaa 1740ctgcccaaag cagcgggagc caccttaaaa ggaaaactgc atgtcccatt cttgctggca 1800gacggcaaat gcaccgtgcc tctagcacca gaacctatga taaccttcgg tttcagatca 1860gtgtcactga aactgcaccc taagaatccc acatatctaa tcacccgcca acttgctgat 1920gagcctcact acacgcacga gctcatatct gaaccagctg ttaggaattt taccgtcacc 1980gaaaaagggt gggagtttgt atggggaaac cacccgccga aaaggttttg ggcacaggaa 2040acagcacccg gaaatccaca tgggctaccg cacgaggtga taactcatta ttaccacaga 2100taccctatgt ccaccatcct gggtttgtca atttgtgccg ccattgcaac cgtttccgtt 2160gcagcgtcta cctggctgtt ttgcagatca agagttgcgt gcctaactcc ttaccggcta 2220acacctaacg ctaggatacc attttgtctg gctgtgcttt gctgcgcccg cactgcccgg 2280gccgagacca cctgggagtc cttggatcac ctatggaaca ataaccaaca gatgttctgg 2340attcaattgc tgatccctct ggccgccttg atcgtagtga ctcgcctgct caggtgcgtg 2400tgctgtgtcg tgcctttttt agtcatggcc ggcgccgcag gcgccggcgc ctacgagcac 2460gcgaccacga tgccgagcca agcgggaatc tcgtataaca ctatagtcaa cagagcaggc 2520tacgcaccac tccctatcag cataacacca acaaagatca agctgatacc tacagtgaac 2580ttggagtacg tcacctgcca ctacaaaaca ggaatggatt caccagccat caaatgctgc 2640ggatctcagg aatgcactcc aacttacagg cctgatgaac agtgcaaagt cttcacaggg 2700gtttacccgt tcatgtgggg tggtgcatat tgcttttgcg acactgagaa cacccaagtc 2760agcaaggcct acgtaatgaa atctgacgac tgccttgcgg atcatgctga agcatataaa

2820gcgcacacag cctcagtgca ggcgttcctc aacatcacag tgggagaaca ctctattgtg 2880actaccgtgt atgtgaatgg agaaactcct gtgaatttca atggggtcaa aataactgca 2940ggtccgcttt ccacagcttg gacacccttt gatcgcaaaa tcgtgcagta tgccggggag 3000atctataatt atgattttcc tgagtatggg gcaggacaac caggagcatt tggagatata 3060caatccagaa cagtctcaag ctctgatctg tatgccaata ccaacctagt gctgcagaga 3120cccaaagcag gagcgatcca cgtgccatac actcaggcac cttcgggttt tgagcaatgg 3180aagaaagata aagctccatc attgaaattt accgcccctt tcggatgcga aatatataca 3240aaccccattc gcgccgaaaa ctgtgctgta gggtcaattc cattagcctt tgacattccc 3300gacgccttgt tcaccagggt gtcagaaaca ccgacacttt cagcggccga atgcactctt 3360aacgagtgcg tgtattcttc cgactttggt gggatcgcca cggtcaagta ctcggccagc 3420aagtcaggca agtgcgcagt ccatgtgcca tcagggactg ctaccctaaa agaagcagca 3480gtcgagctaa ccgagcaagg gtcggcgact atccatttct cgaccgcaaa tatccacccg 3540gagttcaggc tccaaatatg cacatcatat gttacgtgca aaggtgattg tcaccccccg 3600aaagaccata ttgtgacaca ccctcagtat cacgcccaaa catttacagc cgcggtgtca 3660aaaaccgcgt ggacgtggtt aacatccctg ctgggaggat cagccgtaat tattataatt 3720ggcttggtgc tggctactat tgtggccatg tacgtgctga ccaaccagaa acataat 3777201259PRTArtificial SequenceSynthetic vector VLP_VEEV VLP 519 -VEEV VLP 20Met Phe Pro Phe Gln Pro Met Tyr Pro Met Gln Pro Met Pro Tyr Arg 1 5 10 15 Asn Pro Phe Ala Ala Pro Arg Arg Pro Trp Phe Pro Arg Thr Asp Pro 20 25 30 Phe Leu Ala Met Gln Val Gln Glu Leu Thr Arg Ser Met Ala Asn Leu 35 40 45 Thr Phe Lys Gln Arg Arg Asp Ala Pro Pro Glu Gly Pro Ser Ala Asn 50 55 60 Lys Pro Lys Lys Glu Ala Ser Gln Lys Gln Lys Gly Gly Gly Gln Gly 65 70 75 80 Lys Lys Lys Lys Asn Gln Gly Lys Lys Lys Ala Lys Thr Gly Pro Pro 85 90 95 Asn Pro Lys Ala Gln Asn Gly Asn Lys Lys Lys Thr Asn Lys Lys Pro 100 105 110 Gly Lys Arg Gln Arg Met Val Met Lys Leu Glu Ser Asp Lys Thr Phe 115 120 125 Pro Ile Met Leu Glu Gly Lys Ile Asn Gly Tyr Ala Cys Val Val Gly 130 135 140 Gly Lys Leu Phe Arg Pro Met His Val Glu Gly Lys Ile Asp Asn Asp 145 150 155 160 Val Leu Ala Ala Leu Lys Thr Lys Lys Ala Ser Lys Tyr Asp Leu Glu 165 170 175 Tyr Ala Asp Val Pro Gln Asn Met Arg Ala Asp Thr Phe Lys Tyr Thr 180 185 190 His Glu Lys Pro Gln Gly Tyr Tyr Ser Trp His His Gly Ala Val Gln 195 200 205 Tyr Glu Asn Gly Arg Phe Thr Val Pro Lys Gly Val Gly Ala Lys Gly 210 215 220 Asp Ser Gly Arg Pro Ile Leu Asp Asn Gln Gly Arg Val Val Ala Ile 225 230 235 240 Val Leu Gly Gly Val Asn Glu Gly Ser Arg Thr Ala Leu Ser Val Val 245 250 255 Met Trp Asn Glu Lys Gly Val Thr Val Lys Tyr Thr Pro Glu Asn Cys 260 265 270 Glu Gln Trp Ser Leu Val Thr Thr Met Cys Leu Leu Ala Asn Val Thr 275 280 285 Phe Pro Cys Ala Gln Pro Pro Ile Cys Tyr Asp Arg Lys Pro Ala Glu 290 295 300 Thr Leu Ala Met Leu Ser Val Asn Val Asp Asn Pro Gly Tyr Asp Glu 305 310 315 320 Leu Leu Glu Ala Ala Val Lys Cys Pro Gly Arg Lys Arg Arg Ser Thr 325 330 335 Glu Glu Leu Phe Asn Glu Tyr Lys Leu Thr Arg Pro Tyr Met Ala Arg 340 345 350 Cys Ile Arg Cys Ala Val Gly Ser Cys His Ser Pro Ile Ala Ile Glu 355 360 365 Ala Val Lys Ser Asp Gly His Asp Gly Tyr Val Arg Leu Gln Thr Ser 370 375 380 Ser Gln Tyr Gly Leu Asp Ser Ser Gly Asn Leu Lys Gly Arg Thr Met 385 390 395 400 Arg Tyr Asp Met His Gly Thr Ile Lys Glu Ile Pro Leu His Gln Val 405 410 415 Ser Leu Tyr Thr Ser Arg Pro Cys His Ile Val Asp Gly His Gly Tyr 420 425 430 Phe Leu Leu Ala Arg Cys Pro Ala Gly Asp Ser Ile Thr Met Glu Phe 435 440 445 Lys Lys Asp Ser Val Arg His Ser Cys Ser Val Pro Tyr Glu Val Lys 450 455 460 Phe Asn Pro Val Gly Arg Glu Leu Tyr Thr His Pro Pro Glu His Gly 465 470 475 480 Val Glu Gln Ala Cys Gln Val Tyr Ala His Asp Ala Gln Asn Arg Gly 485 490 495 Ala Tyr Val Glu Met His Leu Pro Gly Ser Glu Val Asp Ser Ser Leu 500 505 510 Val Ser Leu Ser Gly Ser Ser Gly Gly Ser Ser Val Thr Val Thr Pro 515 520 525 Pro Asp Gly Thr Ser Ala Leu Val Glu Cys Glu Cys Gly Gly Thr Lys 530 535 540 Ile Ser Glu Thr Ile Asn Lys Thr Lys Gln Phe Ser Gln Cys Thr Lys 545 550 555 560 Lys Glu Gln Cys Arg Ala Tyr Arg Leu Gln Asn Asp Lys Trp Val Tyr 565 570 575 Asn Ser Asp Lys Leu Pro Lys Ala Ala Gly Ala Thr Leu Lys Gly Lys 580 585 590 Leu His Val Pro Phe Leu Leu Ala Asp Gly Lys Cys Thr Val Pro Leu 595 600 605 Ala Pro Glu Pro Met Ile Thr Phe Gly Phe Arg Ser Val Ser Leu Lys 610 615 620 Leu His Pro Lys Asn Pro Thr Tyr Leu Ile Thr Arg Gln Leu Ala Asp 625 630 635 640 Glu Pro His Tyr Thr His Glu Leu Ile Ser Glu Pro Ala Val Arg Asn 645 650 655 Phe Thr Val Thr Glu Lys Gly Trp Glu Phe Val Trp Gly Asn His Pro 660 665 670 Pro Lys Arg Phe Trp Ala Gln Glu Thr Ala Pro Gly Asn Pro His Gly 675 680 685 Leu Pro His Glu Val Ile Thr His Tyr Tyr His Arg Tyr Pro Met Ser 690 695 700 Thr Ile Leu Gly Leu Ser Ile Cys Ala Ala Ile Ala Thr Val Ser Val 705 710 715 720 Ala Ala Ser Thr Trp Leu Phe Cys Arg Ser Arg Val Ala Cys Leu Thr 725 730 735 Pro Tyr Arg Leu Thr Pro Asn Ala Arg Ile Pro Phe Cys Leu Ala Val 740 745 750 Leu Cys Cys Ala Arg Thr Ala Arg Ala Glu Thr Thr Trp Glu Ser Leu 755 760 765 Asp His Leu Trp Asn Asn Asn Gln Gln Met Phe Trp Ile Gln Leu Leu 770 775 780 Ile Pro Leu Ala Ala Leu Ile Val Val Thr Arg Leu Leu Arg Cys Val 785 790 795 800 Cys Cys Val Val Pro Phe Leu Val Met Ala Gly Ala Ala Gly Ala Gly 805 810 815 Ala Tyr Glu His Ala Thr Thr Met Pro Ser Gln Ala Gly Ile Ser Tyr 820 825 830 Asn Thr Ile Val Asn Arg Ala Gly Tyr Ala Pro Leu Pro Ile Ser Ile 835 840 845 Thr Pro Thr Lys Ile Lys Leu Ile Pro Thr Val Asn Leu Glu Tyr Val 850 855 860 Thr Cys His Tyr Lys Thr Gly Met Asp Ser Pro Ala Ile Lys Cys Cys 865 870 875 880 Gly Ser Gln Glu Cys Thr Pro Thr Tyr Arg Pro Asp Glu Gln Cys Lys 885 890 895 Val Phe Thr Gly Val Tyr Pro Phe Met Trp Gly Gly Ala Tyr Cys Phe 900 905 910 Cys Asp Thr Glu Asn Thr Gln Val Ser Lys Ala Tyr Val Met Lys Ser 915 920 925 Asp Asp Cys Leu Ala Asp His Ala Glu Ala Tyr Lys Ala His Thr Ala 930 935 940 Ser Val Gln Ala Phe Leu Asn Ile Thr Val Gly Glu His Ser Ile Val 945 950 955 960 Thr Thr Val Tyr Val Asn Gly Glu Thr Pro Val Asn Phe Asn Gly Val 965 970 975 Lys Ile Thr Ala Gly Pro Leu Ser Thr Ala Trp Thr Pro Phe Asp Arg 980 985 990 Lys Ile Val Gln Tyr Ala Gly Glu Ile Tyr Asn Tyr Asp Phe Pro Glu 995 1000 1005 Tyr Gly Ala Gly Gln Pro Gly Ala Phe Gly Asp Ile Gln Ser Arg 1010 1015 1020 Thr Val Ser Ser Ser Asp Leu Tyr Ala Asn Thr Asn Leu Val Leu 1025 1030 1035 Gln Arg Pro Lys Ala Gly Ala Ile His Val Pro Tyr Thr Gln Ala 1040 1045 1050 Pro Ser Gly Phe Glu Gln Trp Lys Lys Asp Lys Ala Pro Ser Leu 1055 1060 1065 Lys Phe Thr Ala Pro Phe Gly Cys Glu Ile Tyr Thr Asn Pro Ile 1070 1075 1080 Arg Ala Glu Asn Cys Ala Val Gly Ser Ile Pro Leu Ala Phe Asp 1085 1090 1095 Ile Pro Asp Ala Leu Phe Thr Arg Val Ser Glu Thr Pro Thr Leu 1100 1105 1110 Ser Ala Ala Glu Cys Thr Leu Asn Glu Cys Val Tyr Ser Ser Asp 1115 1120 1125 Phe Gly Gly Ile Ala Thr Val Lys Tyr Ser Ala Ser Lys Ser Gly 1130 1135 1140 Lys Cys Ala Val His Val Pro Ser Gly Thr Ala Thr Leu Lys Glu 1145 1150 1155 Ala Ala Val Glu Leu Thr Glu Gln Gly Ser Ala Thr Ile His Phe 1160 1165 1170 Ser Thr Ala Asn Ile His Pro Glu Phe Arg Leu Gln Ile Cys Thr 1175 1180 1185 Ser Tyr Val Thr Cys Lys Gly Asp Cys His Pro Pro Lys Asp His 1190 1195 1200 Ile Val Thr His Pro Gln Tyr His Ala Gln Thr Phe Thr Ala Ala 1205 1210 1215 Val Ser Lys Thr Ala Trp Thr Trp Leu Thr Ser Leu Leu Gly Gly 1220 1225 1230 Ser Ala Val Ile Ile Ile Ile Gly Leu Val Leu Ala Thr Ile Val 1235 1240 1245 Ala Met Tyr Val Leu Thr Asn Gln Lys His Asn 1250 1255 213777DNAArtificial SequenceSynthetic vector VLP_VEEV VLP 538 -VEEV VLP 21atgttcccgt tccagccaat gtatccgatg cagccaatgc cctatcgcaa cccgttcgcg 60gccccgcgca ggccctggtt ccccagaacc gacccttttc tggcgatgca ggtgcaggaa 120ttaacccgct cgatggctaa cctgacgttc aagcaacgcc gggacgcgcc acctgagggg 180ccatccgcta ataaaccgaa gaaggaggcc tcgcaaaaac agaaaggggg aggccaaggg 240aagaagaaga agaaccaagg gaagaagaag gctaagacag ggccgcctaa tccgaaggca 300cagaatggaa acaagaagaa gaccaacaag aaaccaggca agagacagcg catggtcatg 360aaattggaat ctgacaagac gttcccaatc atgttggaag ggaagataaa cggctacgct 420tgtgtggtcg gagggaagtt attcaggccg atgcatgtgg aaggcaagat cgacaacgac 480gttctggccg cgcttaagac gaagaaagca tccaaatacg atcttgagta tgcagatgtg 540ccacagaaca tgcgggccga tacattcaaa tacacccatg agaaacccca aggctattac 600agctggcatc atggagcagt ccaatatgaa aatgggcgtt tcacggtgcc gaaaggagtt 660ggggccaagg gagacagcgg acgacccatt ctggataacc agggacgggt ggtcgctatt 720gtgctgggag gtgtgaatga aggatctagg acagcccttt cagtcgtcat gtggaacgag 780aagggagtta ccgtgaagta tactccagag aactgcgagc aatggtcact agtgaccacc 840atgtgtctgc tcgccaatgt gacgttccca tgtgctcaac caccaatttg ctacgacaga 900aaaccagcag agactttggc catgctcagc gttaacgttg acaacccggg ctacgatgag 960ctgctggaag cagctgttaa gtgccccgga aggaaaagga gatccaccga ggagctgttt 1020aatgagtata agctaacgcg cccttacatg gccagatgca tcagatgtgc agttgggagc 1080tgccatagtc caatagcaat cgaggcagta aagagcgacg ggcacgacgg ttatgttaga 1140cttcagactt cctcgcagta tggcctggat tcctccggca acttaaaggg caggaccatg 1200cggtatgaca tgcacgggac cattaaagag ataccactac atcaagtgtc actctataca 1260tctcgcccgt gtcacattgt ggatgggcac ggttatttcc tgcttgccag gtgcccggca 1320ggggactcca tcaccatgga atttaagaaa gattccgtca gacactcctg ctcggtgccg 1380tatgaagtga aatttaatcc tgtaggcaga gaactctata ctcatccccc agaacacgga 1440gtagagcaag cgtgccaagt ctacgcacat gatgcacaga acagaggagc ttatgtcgag 1500atgcacctcc cgggctcaga agtggacagc agtttggttt ccttgagcgg cagttcagtc 1560accgtgacac ctcctgatgg gactagcgcc ctggtggaat gcgagtgtgg ctccggagga 1620tccggcacaa agatctccga gaccatcaac aagacaaaac agttcagcca gtgcacaaag 1680aaggagcagt gcagagcata tcggctgcag aacgataagt gggtgtataa ttctgacaaa 1740ctgcccaaag cagcgggagc caccttaaaa ggaaaactgc atgtcccatt cttgctggca 1800gacggcaaat gcaccgtgcc tctagcacca gaacctatga taaccttcgg tttcagatca 1860gtgtcactga aactgcaccc taagaatccc acatatctaa tcacccgcca acttgctgat 1920gagcctcact acacgcacga gctcatatct gaaccagctg ttaggaattt taccgtcacc 1980gaaaaagggt gggagtttgt atggggaaac cacccgccga aaaggttttg ggcacaggaa 2040acagcacccg gaaatccaca tgggctaccg cacgaggtga taactcatta ttaccacaga 2100taccctatgt ccaccatcct gggtttgtca atttgtgccg ccattgcaac cgtttccgtt 2160gcagcgtcta cctggctgtt ttgcagatca agagttgcgt gcctaactcc ttaccggcta 2220acacctaacg ctaggatacc attttgtctg gctgtgcttt gctgcgcccg cactgcccgg 2280gccgagacca cctgggagtc cttggatcac ctatggaaca ataaccaaca gatgttctgg 2340attcaattgc tgatccctct ggccgccttg atcgtagtga ctcgcctgct caggtgcgtg 2400tgctgtgtcg tgcctttttt agtcatggcc ggcgccgcag gcgccggcgc ctacgagcac 2460gcgaccacga tgccgagcca agcgggaatc tcgtataaca ctatagtcaa cagagcaggc 2520tacgcaccac tccctatcag cataacacca acaaagatca agctgatacc tacagtgaac 2580ttggagtacg tcacctgcca ctacaaaaca ggaatggatt caccagccat caaatgctgc 2640ggatctcagg aatgcactcc aacttacagg cctgatgaac agtgcaaagt cttcacaggg 2700gtttacccgt tcatgtgggg tggtgcatat tgcttttgcg acactgagaa cacccaagtc 2760agcaaggcct acgtaatgaa atctgacgac tgccttgcgg atcatgctga agcatataaa 2820gcgcacacag cctcagtgca ggcgttcctc aacatcacag tgggagaaca ctctattgtg 2880actaccgtgt atgtgaatgg agaaactcct gtgaatttca atggggtcaa aataactgca 2940ggtccgcttt ccacagcttg gacacccttt gatcgcaaaa tcgtgcagta tgccggggag 3000atctataatt atgattttcc tgagtatggg gcaggacaac caggagcatt tggagatata 3060caatccagaa cagtctcaag ctctgatctg tatgccaata ccaacctagt gctgcagaga 3120cccaaagcag gagcgatcca cgtgccatac actcaggcac cttcgggttt tgagcaatgg 3180aagaaagata aagctccatc attgaaattt accgcccctt tcggatgcga aatatataca 3240aaccccattc gcgccgaaaa ctgtgctgta gggtcaattc cattagcctt tgacattccc 3300gacgccttgt tcaccagggt gtcagaaaca ccgacacttt cagcggccga atgcactctt 3360aacgagtgcg tgtattcttc cgactttggt gggatcgcca cggtcaagta ctcggccagc 3420aagtcaggca agtgcgcagt ccatgtgcca tcagggactg ctaccctaaa agaagcagca 3480gtcgagctaa ccgagcaagg gtcggcgact atccatttct cgaccgcaaa tatccacccg 3540gagttcaggc tccaaatatg cacatcatat gttacgtgca aaggtgattg tcaccccccg 3600aaagaccata ttgtgacaca ccctcagtat cacgcccaaa catttacagc cgcggtgtca 3660aaaaccgcgt ggacgtggtt aacatccctg ctgggaggat cagccgtaat tattataatt 3720ggcttggtgc tggctactat tgtggccatg tacgtgctga ccaaccagaa acataat 3777221259PRTArtificial SequenceSynthetic vector VLP_VEEV VLP 538 -VEEV VLP 22Met Phe Pro Phe Gln Pro Met Tyr Pro Met Gln Pro Met Pro Tyr Arg 1 5 10 15 Asn Pro Phe Ala Ala Pro Arg Arg Pro Trp Phe Pro Arg Thr Asp Pro 20 25 30 Phe Leu Ala Met Gln Val Gln Glu Leu Thr Arg Ser Met Ala Asn Leu 35 40 45 Thr Phe Lys Gln Arg Arg Asp Ala Pro Pro Glu Gly Pro Ser Ala Asn 50 55 60 Lys Pro Lys Lys Glu Ala Ser Gln Lys Gln Lys Gly Gly Gly Gln Gly 65 70 75 80 Lys Lys Lys Lys Asn Gln Gly Lys Lys Lys Ala Lys Thr Gly Pro Pro 85 90 95 Asn Pro Lys Ala Gln Asn Gly Asn Lys Lys Lys Thr Asn Lys Lys Pro 100 105 110 Gly Lys Arg Gln Arg Met Val Met Lys Leu Glu Ser Asp Lys Thr Phe 115 120 125 Pro Ile Met Leu Glu Gly Lys Ile Asn Gly Tyr Ala Cys Val Val Gly 130 135 140 Gly Lys Leu Phe Arg Pro Met His Val Glu Gly Lys Ile Asp Asn Asp 145 150 155 160 Val Leu Ala Ala Leu Lys Thr Lys Lys Ala Ser Lys Tyr Asp Leu Glu 165 170 175 Tyr Ala Asp Val Pro Gln Asn Met Arg Ala Asp Thr Phe Lys Tyr Thr 180 185 190 His Glu Lys Pro Gln Gly Tyr Tyr Ser Trp His His Gly Ala Val Gln 195 200 205 Tyr Glu Asn Gly Arg Phe Thr Val Pro Lys Gly Val Gly Ala Lys Gly 210 215 220 Asp Ser Gly Arg Pro Ile Leu Asp Asn Gln Gly Arg Val Val Ala Ile 225 230 235 240 Val Leu Gly Gly Val Asn Glu Gly Ser Arg Thr Ala Leu Ser Val Val 245 250 255 Met Trp Asn Glu Lys Gly Val Thr Val Lys Tyr Thr Pro Glu Asn Cys 260 265 270 Glu Gln Trp Ser Leu Val Thr

Thr Met Cys Leu Leu Ala Asn Val Thr 275 280 285 Phe Pro Cys Ala Gln Pro Pro Ile Cys Tyr Asp Arg Lys Pro Ala Glu 290 295 300 Thr Leu Ala Met Leu Ser Val Asn Val Asp Asn Pro Gly Tyr Asp Glu 305 310 315 320 Leu Leu Glu Ala Ala Val Lys Cys Pro Gly Arg Lys Arg Arg Ser Thr 325 330 335 Glu Glu Leu Phe Asn Glu Tyr Lys Leu Thr Arg Pro Tyr Met Ala Arg 340 345 350 Cys Ile Arg Cys Ala Val Gly Ser Cys His Ser Pro Ile Ala Ile Glu 355 360 365 Ala Val Lys Ser Asp Gly His Asp Gly Tyr Val Arg Leu Gln Thr Ser 370 375 380 Ser Gln Tyr Gly Leu Asp Ser Ser Gly Asn Leu Lys Gly Arg Thr Met 385 390 395 400 Arg Tyr Asp Met His Gly Thr Ile Lys Glu Ile Pro Leu His Gln Val 405 410 415 Ser Leu Tyr Thr Ser Arg Pro Cys His Ile Val Asp Gly His Gly Tyr 420 425 430 Phe Leu Leu Ala Arg Cys Pro Ala Gly Asp Ser Ile Thr Met Glu Phe 435 440 445 Lys Lys Asp Ser Val Arg His Ser Cys Ser Val Pro Tyr Glu Val Lys 450 455 460 Phe Asn Pro Val Gly Arg Glu Leu Tyr Thr His Pro Pro Glu His Gly 465 470 475 480 Val Glu Gln Ala Cys Gln Val Tyr Ala His Asp Ala Gln Asn Arg Gly 485 490 495 Ala Tyr Val Glu Met His Leu Pro Gly Ser Glu Val Asp Ser Ser Leu 500 505 510 Val Ser Leu Ser Gly Ser Ser Val Thr Val Thr Pro Pro Asp Gly Thr 515 520 525 Ser Ala Leu Val Glu Cys Glu Cys Gly Ser Gly Gly Ser Gly Thr Lys 530 535 540 Ile Ser Glu Thr Ile Asn Lys Thr Lys Gln Phe Ser Gln Cys Thr Lys 545 550 555 560 Lys Glu Gln Cys Arg Ala Tyr Arg Leu Gln Asn Asp Lys Trp Val Tyr 565 570 575 Asn Ser Asp Lys Leu Pro Lys Ala Ala Gly Ala Thr Leu Lys Gly Lys 580 585 590 Leu His Val Pro Phe Leu Leu Ala Asp Gly Lys Cys Thr Val Pro Leu 595 600 605 Ala Pro Glu Pro Met Ile Thr Phe Gly Phe Arg Ser Val Ser Leu Lys 610 615 620 Leu His Pro Lys Asn Pro Thr Tyr Leu Ile Thr Arg Gln Leu Ala Asp 625 630 635 640 Glu Pro His Tyr Thr His Glu Leu Ile Ser Glu Pro Ala Val Arg Asn 645 650 655 Phe Thr Val Thr Glu Lys Gly Trp Glu Phe Val Trp Gly Asn His Pro 660 665 670 Pro Lys Arg Phe Trp Ala Gln Glu Thr Ala Pro Gly Asn Pro His Gly 675 680 685 Leu Pro His Glu Val Ile Thr His Tyr Tyr His Arg Tyr Pro Met Ser 690 695 700 Thr Ile Leu Gly Leu Ser Ile Cys Ala Ala Ile Ala Thr Val Ser Val 705 710 715 720 Ala Ala Ser Thr Trp Leu Phe Cys Arg Ser Arg Val Ala Cys Leu Thr 725 730 735 Pro Tyr Arg Leu Thr Pro Asn Ala Arg Ile Pro Phe Cys Leu Ala Val 740 745 750 Leu Cys Cys Ala Arg Thr Ala Arg Ala Glu Thr Thr Trp Glu Ser Leu 755 760 765 Asp His Leu Trp Asn Asn Asn Gln Gln Met Phe Trp Ile Gln Leu Leu 770 775 780 Ile Pro Leu Ala Ala Leu Ile Val Val Thr Arg Leu Leu Arg Cys Val 785 790 795 800 Cys Cys Val Val Pro Phe Leu Val Met Ala Gly Ala Ala Gly Ala Gly 805 810 815 Ala Tyr Glu His Ala Thr Thr Met Pro Ser Gln Ala Gly Ile Ser Tyr 820 825 830 Asn Thr Ile Val Asn Arg Ala Gly Tyr Ala Pro Leu Pro Ile Ser Ile 835 840 845 Thr Pro Thr Lys Ile Lys Leu Ile Pro Thr Val Asn Leu Glu Tyr Val 850 855 860 Thr Cys His Tyr Lys Thr Gly Met Asp Ser Pro Ala Ile Lys Cys Cys 865 870 875 880 Gly Ser Gln Glu Cys Thr Pro Thr Tyr Arg Pro Asp Glu Gln Cys Lys 885 890 895 Val Phe Thr Gly Val Tyr Pro Phe Met Trp Gly Gly Ala Tyr Cys Phe 900 905 910 Cys Asp Thr Glu Asn Thr Gln Val Ser Lys Ala Tyr Val Met Lys Ser 915 920 925 Asp Asp Cys Leu Ala Asp His Ala Glu Ala Tyr Lys Ala His Thr Ala 930 935 940 Ser Val Gln Ala Phe Leu Asn Ile Thr Val Gly Glu His Ser Ile Val 945 950 955 960 Thr Thr Val Tyr Val Asn Gly Glu Thr Pro Val Asn Phe Asn Gly Val 965 970 975 Lys Ile Thr Ala Gly Pro Leu Ser Thr Ala Trp Thr Pro Phe Asp Arg 980 985 990 Lys Ile Val Gln Tyr Ala Gly Glu Ile Tyr Asn Tyr Asp Phe Pro Glu 995 1000 1005 Tyr Gly Ala Gly Gln Pro Gly Ala Phe Gly Asp Ile Gln Ser Arg 1010 1015 1020 Thr Val Ser Ser Ser Asp Leu Tyr Ala Asn Thr Asn Leu Val Leu 1025 1030 1035 Gln Arg Pro Lys Ala Gly Ala Ile His Val Pro Tyr Thr Gln Ala 1040 1045 1050 Pro Ser Gly Phe Glu Gln Trp Lys Lys Asp Lys Ala Pro Ser Leu 1055 1060 1065 Lys Phe Thr Ala Pro Phe Gly Cys Glu Ile Tyr Thr Asn Pro Ile 1070 1075 1080 Arg Ala Glu Asn Cys Ala Val Gly Ser Ile Pro Leu Ala Phe Asp 1085 1090 1095 Ile Pro Asp Ala Leu Phe Thr Arg Val Ser Glu Thr Pro Thr Leu 1100 1105 1110 Ser Ala Ala Glu Cys Thr Leu Asn Glu Cys Val Tyr Ser Ser Asp 1115 1120 1125 Phe Gly Gly Ile Ala Thr Val Lys Tyr Ser Ala Ser Lys Ser Gly 1130 1135 1140 Lys Cys Ala Val His Val Pro Ser Gly Thr Ala Thr Leu Lys Glu 1145 1150 1155 Ala Ala Val Glu Leu Thr Glu Gln Gly Ser Ala Thr Ile His Phe 1160 1165 1170 Ser Thr Ala Asn Ile His Pro Glu Phe Arg Leu Gln Ile Cys Thr 1175 1180 1185 Ser Tyr Val Thr Cys Lys Gly Asp Cys His Pro Pro Lys Asp His 1190 1195 1200 Ile Val Thr His Pro Gln Tyr His Ala Gln Thr Phe Thr Ala Ala 1205 1210 1215 Val Ser Lys Thr Ala Trp Thr Trp Leu Thr Ser Leu Leu Gly Gly 1220 1225 1230 Ser Ala Val Ile Ile Ile Ile Gly Leu Val Leu Ala Thr Ile Val 1235 1240 1245 Ala Met Tyr Val Leu Thr Asn Gln Lys His Asn 1250 1255 238416DNAArtificial SequenceSynthetic vector VLP_CHI 520 23gaattcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt 60ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg 120gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 180ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 240atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 300gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 360gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 420tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 480atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 540gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 600tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga 660gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 720agaagacacc gggaccgatc cagcctccgt taacggtgga gggcagtgta gtctgagcag 780tactcgttgc tgccgcgcgc gccaccagac ataatagctg acagactaac agactgttcc 840tttccatggg tcttttctgc agtcaccgtc gtcgacacgt gtgatcagat atcgcggccg 900ccaccatgga gttcatcccg acgcaaactt tctataacag aaggtaccaa ccccgaccct 960gggccccacg ccctacaatt caagtaatta gacctagacc acgtccacag aggcaggctg 1020ggcaactcgc ccagctgatc tccgcagtca acaaattgac catgcgcgcg gtacctcaac 1080agaagcctcg cagaaatcgg aaaaacaaga agcaaaggca gaagaagcag gcgccgcaaa 1140acgacccaaa gcaaaagaag caaccaccac aaaagaagcc ggctcaaaag aagaagaaac 1200caggccgtag ggagagaatg tgcatgaaaa ttgaaaatga ttgcatcttc gaagtcaagc 1260atgaaggcaa agtgatgggc tacgcatgcc tggtggggga taaagtaatg aaaccagcac 1320atgtgaaggg aactatcgac aatgccgatc tggctaaact ggcctttaag cggtcgtcta 1380aatacgatct tgaatgtgca cagataccgg tgcacatgaa gtctgatgcc tcgaagttta 1440cccacgagaa acccgagggg tactataact ggcatcacgg agcagtgcag tattcaggag 1500gccggttcac tatcccgacg ggtgcaggca agccgggaga cagcggcaga ccgatcttcg 1560acaacaaagg acgggtggtg gccatcgtcc taggaggggc caacgaaggt gcccgcacgg 1620ccctctccgt ggtgacgtgg aacaaagaca tcgtcacaaa aattacccct gagggagccg 1680aagagtggag cctcgccctc ccggtcttgt gcctgttggc aaacactaca ttcccctgct 1740ctcagccgcc ttgcacaccc tgctgctacg aaaaggaacc ggaaagcacc ttgcgcatgc 1800ttgaggacaa cgtgatgaga cccggatact accagctact aaaagcatcg ctgacttgct 1860ctccccaccg ccaaagacgc agtactaagg acaattttaa tgtctataaa gccacaagac 1920catatctagc tcattgtcct gactgcggag aagggcattc gtgccacagc cctatcgcat 1980tggagcgcat cagaaatgaa gcaacggacg gaacgctgaa aatccaggtc tctttgcaga 2040tcgggataaa gacagatgac agccacgatt ggaccaagct gcgctatatg gatagccata 2100cgccagcgga cgcggagcga gccggattgc ttgtaaggac ttcagcaccg tgcacgatca 2160ccgggaccat gggacacttt attctcgccc gatgcccgaa aggagagacg ctgacagtgg 2220gatttacgga cagcagaaag atcagccaca catgcacaca cccgttccat catgaaccac 2280ctgtgatagg tagggagagg ttccactctc gaccacaaca tggtaaagag ttaccttgca 2340gcacgtacgt gcagagcacc gctgccactg ctgaggagat agaggtgcat atgcccccag 2400atactcctga ccgcacgctg atgacgcagc agtctggcaa cgtgaagatc acagttaatg 2460gctccggagg atccggcggg cagacggtgc ggtacaagtg caactgcggt ggctcaaacg 2520agggactgac aaccacagac aaagtgatca ataactgcaa aattgatcag tgccatgctg 2580cagtcactaa tcacaagaat tggcaataca actccccttt agtcccgcgc aacgctgaac 2640tcggggaccg taaaggaaag atccacatcc cattcccatt ggcaaacgtg acttgcagag 2700tgccaaaagc aagaaaccct acagtaactt acggaaaaaa ccaagtcacc atgctgctgt 2760atcctgacca tccgacactc ttgtcttacc gtaacatggg acaggaacca aattaccacg 2820aggagtgggt gacacacaag aaggaggtta ccttgaccgt gcctactgag ggtctggagg 2880tcacttgggg caacaacgaa ccatacaagt actggccgca gatgtctacg aacggtactg 2940ctcatggtca cccacatgag ataatcttgt actattatga gctgtacccc actatgactg 3000tagtcattgt gtcggtggcc tcgttcgtgc ttctgtcgat ggtgggcaca gcagtgggaa 3060tgtgtgtgtg cgcacggcgc agatgcatta caccatatga attaacacca ggagccactg 3120ttcccttcct gctcagcctg ctatgctgcg tcagaacgac caaggcggcc acatattacg 3180aggctgcggc atatctatgg aacgaacagc agcccctgtt ctggttgcag gctcttatcc 3240cgctggccgc cttgatcgtc ctgtgcaact gtctgaaact cttgccatgc tgctgtaaga 3300ccctggcttt tttagccgta atgagcatcg gtgcccacac tgtgagcgcg tacgaacacg 3360taacagtgat cccgaacacg gtgggagtac cgtataagac tcttgtcaac agaccgggtt 3420acagccccat ggtgttggag atggagctac aatcagtcac cttggaacca acactgtcac 3480ttgactacat cacgtgcgag tacaaaactg tcatcccctc cccgtacgtg aagtgctgtg 3540gtacagcaga gtgcaaggac aagagcctac cagactacag ctgcaaggtc tttactggag 3600tctacccatt tatgtggggc ggcgcctact gcttttgcga cgccgaaaat acgcaattga 3660gcgaggcaca tgtagagaaa tctgaatctt gcaaaacaga gtttgcatcg gcctacagag 3720cccacaccgc atcggcgtcg gcgaagctcc gcgtccttta ccaaggaaac aacattaccg 3780tagctgccta cgctaacggt gaccatgccg tcacagtaaa ggacgccaag tttgtcgtgg 3840gcccaatgtc ctccgcctgg acaccttttg acaacaaaat cgtggtgtac aaaggcgacg 3900tctacaacat ggactaccca ccttttggcg caggaagacc aggacaattt ggtgacattc 3960aaagtcgtac accggaaagt aaagacgttt atgccaacac tcagttggta ctacagaggc 4020cagcagcagg cacggtacat gtaccatact ctcaggcacc atctggcttc aagtattggc 4080tgaaggaacg aggagcatcg ctacagcaca cggcaccgtt cggttgccag attgcgacaa 4140acccggtaag agctgtaaat tgcgctgtgg ggaacatacc aatttccatc gacataccgg 4200atgcggcctt tactagggtt gtcgatgcac cctctgtaac ggacatgtca tgcgaagtac 4260cagcctgcac tcactcctcc gactttgggg gcgtcgccat catcaaatac acagctagca 4320agaaaggtaa atgtgcagta cattcgatga ccaacgccgt taccattcga gaagccgacg 4380tagaagtaga ggggaactcc cagctgcaaa tatccttctc aacagccctg gcaagcgccg 4440agtttcgcgt gcaagtgtgc tccacacaag tacactgcgc agccgcatgc caccctccaa 4500aggaccacat agtcaattac ccagcatcac acaccaccct tggggtccag gatatatcca 4560caacggcaat gtcttgggtg cagaagatta cgggaggagt aggattaatt gttgctgttg 4620ctgccttaat tttaattgtg gtgctatgcg tgtcgtttag caggcactaa ggatctagat 4680ctgctgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga 4740ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt 4800gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg 4860attgggaaga caatagcagg catgctgggg atgcggtggg ctctatgggt acccaggtgc 4920tgaagaattg acccggttcc tcctgggcca gaaagaagca ggcacatccc cttctctgtg 4980acacaccctg tccacgcccc tggttcttag ttccagcccc actcatagga cactcatagc 5040tcaggagggc tccgccttca atcccacccg ctaaagtact tggagcggtc tctccctccc 5100tcatcagccc accaaaccaa acctagcctc caagagtggg aagaaattaa agcaagatag 5160gctattaagt gcagagggag agaaaatgcc tccaacatgt gaggaagtaa tgagagaaat 5220catagaattt taaggccatg atttaaggcc atcatggcct aagcttgaaa ggagatagga 5280tcaaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 5340aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 5400gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 5460gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 5520ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 5580atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 5640gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 5700gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 5760gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 5820gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 5880aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 5940ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 6000taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 6060tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 6120gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 6180taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 6240tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 6300tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 6360ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 6420taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 6480tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 6540cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 6600gcgagaacca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 6660cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 6720ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 6780aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 6840atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 6900tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 6960gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 7020aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 7080acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 7140ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 7200tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 7260aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 7320catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 7380atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 7440aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 7500gcgtatcacg aggccctttc gggtcgcgcg tttcggtgat gacggtgaaa acctctgaca 7560catgcagctc ccgttgacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 7620ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 7680agagcagatt gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt 7740cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat 7800cccttataaa tcaaaagaat agcccgagat agggttgagt gttgttccag tttggaacaa 7860gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg 7920cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa 7980agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc 8040gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag 8100tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg 8160cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga 8220aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg 8280gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta 8340agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattc 8400catggtctca actttc 8416243762DNAArtificial

SequenceSynthetic vector insert VLP_CHI 520 24atggagttca tcccgacgca aactttctat aacagaaggt accaaccccg accctgggcc 60ccacgcccta caattcaagt aattagacct agaccacgtc cacagaggca ggctgggcaa 120ctcgcccagc tgatctccgc agtcaacaaa ttgaccatgc gcgcggtacc tcaacagaag 180cctcgcagaa atcggaaaaa caagaagcaa aggcagaaga agcaggcgcc gcaaaacgac 240ccaaagcaaa agaagcaacc accacaaaag aagccggctc aaaagaagaa gaaaccaggc 300cgtagggaga gaatgtgcat gaaaattgaa aatgattgca tcttcgaagt caagcatgaa 360ggcaaagtga tgggctacgc atgcctggtg ggggataaag taatgaaacc agcacatgtg 420aagggaacta tcgacaatgc cgatctggct aaactggcct ttaagcggtc gtctaaatac 480gatcttgaat gtgcacagat accggtgcac atgaagtctg atgcctcgaa gtttacccac 540gagaaacccg aggggtacta taactggcat cacggagcag tgcagtattc aggaggccgg 600ttcactatcc cgacgggtgc aggcaagccg ggagacagcg gcagaccgat cttcgacaac 660aaaggacggg tggtggccat cgtcctagga ggggccaacg aaggtgcccg cacggccctc 720tccgtggtga cgtggaacaa agacatcgtc acaaaaatta cccctgaggg agccgaagag 780tggagcctcg ccctcccggt cttgtgcctg ttggcaaaca ctacattccc ctgctctcag 840ccgccttgca caccctgctg ctacgaaaag gaaccggaaa gcaccttgcg catgcttgag 900gacaacgtga tgagacccgg atactaccag ctactaaaag catcgctgac ttgctctccc 960caccgccaaa gacgcagtac taaggacaat tttaatgtct ataaagccac aagaccatat 1020ctagctcatt gtcctgactg cggagaaggg cattcgtgcc acagccctat cgcattggag 1080cgcatcagaa atgaagcaac ggacggaacg ctgaaaatcc aggtctcttt gcagatcggg 1140ataaagacag atgacagcca cgattggacc aagctgcgct atatggatag ccatacgcca 1200gcggacgcgg agcgagccgg attgcttgta aggacttcag caccgtgcac gatcaccggg 1260accatgggac actttattct cgcccgatgc ccgaaaggag agacgctgac agtgggattt 1320acggacagca gaaagatcag ccacacatgc acacacccgt tccatcatga accacctgtg 1380ataggtaggg agaggttcca ctctcgacca caacatggta aagagttacc ttgcagcacg 1440tacgtgcaga gcaccgctgc cactgctgag gagatagagg tgcatatgcc cccagatact 1500cctgaccgca cgctgatgac gcagcagtct ggcaacgtga agatcacagt taatggctcc 1560ggaggatccg gcgggcagac ggtgcggtac aagtgcaact gcggtggctc aaacgaggga 1620ctgacaacca cagacaaagt gatcaataac tgcaaaattg atcagtgcca tgctgcagtc 1680actaatcaca agaattggca atacaactcc cctttagtcc cgcgcaacgc tgaactcggg 1740gaccgtaaag gaaagatcca catcccattc ccattggcaa acgtgacttg cagagtgcca 1800aaagcaagaa accctacagt aacttacgga aaaaaccaag tcaccatgct gctgtatcct 1860gaccatccga cactcttgtc ttaccgtaac atgggacagg aaccaaatta ccacgaggag 1920tgggtgacac acaagaagga ggttaccttg accgtgccta ctgagggtct ggaggtcact 1980tggggcaaca acgaaccata caagtactgg ccgcagatgt ctacgaacgg tactgctcat 2040ggtcacccac atgagataat cttgtactat tatgagctgt accccactat gactgtagtc 2100attgtgtcgg tggcctcgtt cgtgcttctg tcgatggtgg gcacagcagt gggaatgtgt 2160gtgtgcgcac ggcgcagatg cattacacca tatgaattaa caccaggagc cactgttccc 2220ttcctgctca gcctgctatg ctgcgtcaga acgaccaagg cggccacata ttacgaggct 2280gcggcatatc tatggaacga acagcagccc ctgttctggt tgcaggctct tatcccgctg 2340gccgccttga tcgtcctgtg caactgtctg aaactcttgc catgctgctg taagaccctg 2400gcttttttag ccgtaatgag catcggtgcc cacactgtga gcgcgtacga acacgtaaca 2460gtgatcccga acacggtggg agtaccgtat aagactcttg tcaacagacc gggttacagc 2520cccatggtgt tggagatgga gctacaatca gtcaccttgg aaccaacact gtcacttgac 2580tacatcacgt gcgagtacaa aactgtcatc ccctccccgt acgtgaagtg ctgtggtaca 2640gcagagtgca aggacaagag cctaccagac tacagctgca aggtctttac tggagtctac 2700ccatttatgt ggggcggcgc ctactgcttt tgcgacgccg aaaatacgca attgagcgag 2760gcacatgtag agaaatctga atcttgcaaa acagagtttg catcggccta cagagcccac 2820accgcatcgg cgtcggcgaa gctccgcgtc ctttaccaag gaaacaacat taccgtagct 2880gcctacgcta acggtgacca tgccgtcaca gtaaaggacg ccaagtttgt cgtgggccca 2940atgtcctccg cctggacacc ttttgacaac aaaatcgtgg tgtacaaagg cgacgtctac 3000aacatggact acccaccttt tggcgcagga agaccaggac aatttggtga cattcaaagt 3060cgtacaccgg aaagtaaaga cgtttatgcc aacactcagt tggtactaca gaggccagca 3120gcaggcacgg tacatgtacc atactctcag gcaccatctg gcttcaagta ttggctgaag 3180gaacgaggag catcgctaca gcacacggca ccgttcggtt gccagattgc gacaaacccg 3240gtaagagctg taaattgcgc tgtggggaac ataccaattt ccatcgacat accggatgcg 3300gcctttacta gggttgtcga tgcaccctct gtaacggaca tgtcatgcga agtaccagcc 3360tgcactcact cctccgactt tgggggcgtc gccatcatca aatacacagc tagcaagaaa 3420ggtaaatgtg cagtacattc gatgaccaac gccgttacca ttcgagaagc cgacgtagaa 3480gtagagggga actcccagct gcaaatatcc ttctcaacag ccctggcaag cgccgagttt 3540cgcgtgcaag tgtgctccac acaagtacac tgcgcagccg catgccaccc tccaaaggac 3600cacatagtca attacccagc atcacacacc acccttgggg tccaggatat atccacaacg 3660gcaatgtctt gggtgcagaa gattacggga ggagtaggat taattgttgc tgttgctgcc 3720ttaattttaa ttgtggtgct atgcgtgtcg tttagcaggc ac 3762251254PRTArtificial SequenceSynthetic vector VLP_CHI 520 25Met Glu Phe Ile Pro Thr Gln Thr Phe Tyr Asn Arg Arg Tyr Gln Pro 1 5 10 15 Arg Pro Trp Ala Pro Arg Pro Thr Ile Gln Val Ile Arg Pro Arg Pro 20 25 30 Arg Pro Gln Arg Gln Ala Gly Gln Leu Ala Gln Leu Ile Ser Ala Val 35 40 45 Asn Lys Leu Thr Met Arg Ala Val Pro Gln Gln Lys Pro Arg Arg Asn 50 55 60 Arg Lys Asn Lys Lys Gln Arg Gln Lys Lys Gln Ala Pro Gln Asn Asp 65 70 75 80 Pro Lys Gln Lys Lys Gln Pro Pro Gln Lys Lys Pro Ala Gln Lys Lys 85 90 95 Lys Lys Pro Gly Arg Arg Glu Arg Met Cys Met Lys Ile Glu Asn Asp 100 105 110 Cys Ile Phe Glu Val Lys His Glu Gly Lys Val Met Gly Tyr Ala Cys 115 120 125 Leu Val Gly Asp Lys Val Met Lys Pro Ala His Val Lys Gly Thr Ile 130 135 140 Asp Asn Ala Asp Leu Ala Lys Leu Ala Phe Lys Arg Ser Ser Lys Tyr 145 150 155 160 Asp Leu Glu Cys Ala Gln Ile Pro Val His Met Lys Ser Asp Ala Ser 165 170 175 Lys Phe Thr His Glu Lys Pro Glu Gly Tyr Tyr Asn Trp His His Gly 180 185 190 Ala Val Gln Tyr Ser Gly Gly Arg Phe Thr Ile Pro Thr Gly Ala Gly 195 200 205 Lys Pro Gly Asp Ser Gly Arg Pro Ile Phe Asp Asn Lys Gly Arg Val 210 215 220 Val Ala Ile Val Leu Gly Gly Ala Asn Glu Gly Ala Arg Thr Ala Leu 225 230 235 240 Ser Val Val Thr Trp Asn Lys Asp Ile Val Thr Lys Ile Thr Pro Glu 245 250 255 Gly Ala Glu Glu Trp Ser Leu Ala Leu Pro Val Leu Cys Leu Leu Ala 260 265 270 Asn Thr Thr Phe Pro Cys Ser Gln Pro Pro Cys Thr Pro Cys Cys Tyr 275 280 285 Glu Lys Glu Pro Glu Ser Thr Leu Arg Met Leu Glu Asp Asn Val Met 290 295 300 Arg Pro Gly Tyr Tyr Gln Leu Leu Lys Ala Ser Leu Thr Cys Ser Pro 305 310 315 320 His Arg Gln Arg Arg Ser Thr Lys Asp Asn Phe Asn Val Tyr Lys Ala 325 330 335 Thr Arg Pro Tyr Leu Ala His Cys Pro Asp Cys Gly Glu Gly His Ser 340 345 350 Cys His Ser Pro Ile Ala Leu Glu Arg Ile Arg Asn Glu Ala Thr Asp 355 360 365 Gly Thr Leu Lys Ile Gln Val Ser Leu Gln Ile Gly Ile Lys Thr Asp 370 375 380 Asp Ser His Asp Trp Thr Lys Leu Arg Tyr Met Asp Ser His Thr Pro 385 390 395 400 Ala Asp Ala Glu Arg Ala Gly Leu Leu Val Arg Thr Ser Ala Pro Cys 405 410 415 Thr Ile Thr Gly Thr Met Gly His Phe Ile Leu Ala Arg Cys Pro Lys 420 425 430 Gly Glu Thr Leu Thr Val Gly Phe Thr Asp Ser Arg Lys Ile Ser His 435 440 445 Thr Cys Thr His Pro Phe His His Glu Pro Pro Val Ile Gly Arg Glu 450 455 460 Arg Phe His Ser Arg Pro Gln His Gly Lys Glu Leu Pro Cys Ser Thr 465 470 475 480 Tyr Val Gln Ser Thr Ala Ala Thr Ala Glu Glu Ile Glu Val His Met 485 490 495 Pro Pro Asp Thr Pro Asp Arg Thr Leu Met Thr Gln Gln Ser Gly Asn 500 505 510 Val Lys Ile Thr Val Asn Gly Ser Gly Gly Ser Gly Gly Gln Thr Val 515 520 525 Arg Tyr Lys Cys Asn Cys Gly Gly Ser Asn Glu Gly Leu Thr Thr Thr 530 535 540 Asp Lys Val Ile Asn Asn Cys Lys Ile Asp Gln Cys His Ala Ala Val 545 550 555 560 Thr Asn His Lys Asn Trp Gln Tyr Asn Ser Pro Leu Val Pro Arg Asn 565 570 575 Ala Glu Leu Gly Asp Arg Lys Gly Lys Ile His Ile Pro Phe Pro Leu 580 585 590 Ala Asn Val Thr Cys Arg Val Pro Lys Ala Arg Asn Pro Thr Val Thr 595 600 605 Tyr Gly Lys Asn Gln Val Thr Met Leu Leu Tyr Pro Asp His Pro Thr 610 615 620 Leu Leu Ser Tyr Arg Asn Met Gly Gln Glu Pro Asn Tyr His Glu Glu 625 630 635 640 Trp Val Thr His Lys Lys Glu Val Thr Leu Thr Val Pro Thr Glu Gly 645 650 655 Leu Glu Val Thr Trp Gly Asn Asn Glu Pro Tyr Lys Tyr Trp Pro Gln 660 665 670 Met Ser Thr Asn Gly Thr Ala His Gly His Pro His Glu Ile Ile Leu 675 680 685 Tyr Tyr Tyr Glu Leu Tyr Pro Thr Met Thr Val Val Ile Val Ser Val 690 695 700 Ala Ser Phe Val Leu Leu Ser Met Val Gly Thr Ala Val Gly Met Cys 705 710 715 720 Val Cys Ala Arg Arg Arg Cys Ile Thr Pro Tyr Glu Leu Thr Pro Gly 725 730 735 Ala Thr Val Pro Phe Leu Leu Ser Leu Leu Cys Cys Val Arg Thr Thr 740 745 750 Lys Ala Ala Thr Tyr Tyr Glu Ala Ala Ala Tyr Leu Trp Asn Glu Gln 755 760 765 Gln Pro Leu Phe Trp Leu Gln Ala Leu Ile Pro Leu Ala Ala Leu Ile 770 775 780 Val Leu Cys Asn Cys Leu Lys Leu Leu Pro Cys Cys Cys Lys Thr Leu 785 790 795 800 Ala Phe Leu Ala Val Met Ser Ile Gly Ala His Thr Val Ser Ala Tyr 805 810 815 Glu His Val Thr Val Ile Pro Asn Thr Val Gly Val Pro Tyr Lys Thr 820 825 830 Leu Val Asn Arg Pro Gly Tyr Ser Pro Met Val Leu Glu Met Glu Leu 835 840 845 Gln Ser Val Thr Leu Glu Pro Thr Leu Ser Leu Asp Tyr Ile Thr Cys 850 855 860 Glu Tyr Lys Thr Val Ile Pro Ser Pro Tyr Val Lys Cys Cys Gly Thr 865 870 875 880 Ala Glu Cys Lys Asp Lys Ser Leu Pro Asp Tyr Ser Cys Lys Val Phe 885 890 895 Thr Gly Val Tyr Pro Phe Met Trp Gly Gly Ala Tyr Cys Phe Cys Asp 900 905 910 Ala Glu Asn Thr Gln Leu Ser Glu Ala His Val Glu Lys Ser Glu Ser 915 920 925 Cys Lys Thr Glu Phe Ala Ser Ala Tyr Arg Ala His Thr Ala Ser Ala 930 935 940 Ser Ala Lys Leu Arg Val Leu Tyr Gln Gly Asn Asn Ile Thr Val Ala 945 950 955 960 Ala Tyr Ala Asn Gly Asp His Ala Val Thr Val Lys Asp Ala Lys Phe 965 970 975 Val Val Gly Pro Met Ser Ser Ala Trp Thr Pro Phe Asp Asn Lys Ile 980 985 990 Val Val Tyr Lys Gly Asp Val Tyr Asn Met Asp Tyr Pro Pro Phe Gly 995 1000 1005 Ala Gly Arg Pro Gly Gln Phe Gly Asp Ile Gln Ser Arg Thr Pro 1010 1015 1020 Glu Ser Lys Asp Val Tyr Ala Asn Thr Gln Leu Val Leu Gln Arg 1025 1030 1035 Pro Ala Ala Gly Thr Val His Val Pro Tyr Ser Gln Ala Pro Ser 1040 1045 1050 Gly Phe Lys Tyr Trp Leu Lys Glu Arg Gly Ala Ser Leu Gln His 1055 1060 1065 Thr Ala Pro Phe Gly Cys Gln Ile Ala Thr Asn Pro Val Arg Ala 1070 1075 1080 Val Asn Cys Ala Val Gly Asn Ile Pro Ile Ser Ile Asp Ile Pro 1085 1090 1095 Asp Ala Ala Phe Thr Arg Val Val Asp Ala Pro Ser Val Thr Asp 1100 1105 1110 Met Ser Cys Glu Val Pro Ala Cys Thr His Ser Ser Asp Phe Gly 1115 1120 1125 Gly Val Ala Ile Ile Lys Tyr Thr Ala Ser Lys Lys Gly Lys Cys 1130 1135 1140 Ala Val His Ser Met Thr Asn Ala Val Thr Ile Arg Glu Ala Asp 1145 1150 1155 Val Glu Val Glu Gly Asn Ser Gln Leu Gln Ile Ser Phe Ser Thr 1160 1165 1170 Ala Leu Ala Ser Ala Glu Phe Arg Val Gln Val Cys Ser Thr Gln 1175 1180 1185 Val His Cys Ala Ala Ala Cys His Pro Pro Lys Asp His Ile Val 1190 1195 1200 Asn Tyr Pro Ala Ser His Thr Thr Leu Gly Val Gln Asp Ile Ser 1205 1210 1215 Thr Thr Ala Met Ser Trp Val Gln Lys Ile Thr Gly Gly Val Gly 1220 1225 1230 Leu Ile Val Ala Val Ala Ala Leu Ile Leu Ile Val Val Leu Cys 1235 1240 1245 Val Ser Phe Ser Arg His 1250 268485DNAArtificial SequenceSynthetic VLP74_11 (CHI VLP 532_NPNAx6) sequence 26gaattcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt 60ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg 120gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 180ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 240atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 300gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 360gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 420tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 480atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 540gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 600tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga 660gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 720agaagacacc gggaccgatc cagcctccgt taacggtgga gggcagtgta gtctgagcag 780tactcgttgc tgccgcgcgc gccaccagac ataatagctg acagactaac agactgttcc 840tttccatggg tcttttctgc agtcaccgtc gtcgacacgt gtgatcagat atcgcggccg 900ccaccatgga gttcatcccg acgcaaactt tctataacag aaggtaccaa ccccgaccct 960gggccccacg ccctacaatt caagtaatta gacctagacc acgtccacag aggcaggctg 1020ggcaactcgc ccagctgatc tccgcagtca acaaattgac catgcgcgcg gtacctcaac 1080agaagcctcg cagaaatcgg aaaaacaaga agcaaaggca gaagaagcag gcgccgcaaa 1140acgacccaaa gcaaaagaag caaccaccac aaaagaagcc ggctcaaaag aagaagaaac 1200caggccgtag ggagagaatg tgcatgaaaa ttgaaaatga ttgcatcttc gaagtcaagc 1260atgaaggcaa agtgatgggc tacgcatgcc tggtggggga taaagtaatg aaaccagcac 1320atgtgaaggg aactatcgac aatgccgatc tggctaaact ggcctttaag cggtcgtcta 1380aatacgatct tgaatgtgca cagataccgg tgcacatgaa gtctgatgcc tcgaagttta 1440cccacgagaa acccgagggg tactataact ggcatcacgg agcagtgcag tattcaggag 1500gccggttcac tatcccgacg ggtgcaggca agccgggaga cagcggcaga ccgatcttcg 1560acaacaaagg acgggtggtg gccatcgtcc taggaggggc caacgaaggt gcccgcacgg 1620ccctctccgt ggtgacgtgg aacaaagaca tcgtcacaaa aattacccct gagggagccg 1680aagagtggag cctcgccctc ccggtcttgt gcctgttggc aaacactaca ttcccctgct 1740ctcagccgcc ttgcacaccc tgctgctacg aaaaggaacc ggaaagcacc ttgcgcatgc 1800ttgaggacaa cgtgatgaga cccggatact accagctact aaaagcatcg ctgacttgct 1860ctccccaccg ccaaagacgc agtactaagg acaattttaa tgtctataaa gccacaagac 1920catatctagc tcattgtcct gactgcggag aagggcattc gtgccacagc cctatcgcat 1980tggagcgcat cagaaatgaa gcaacggacg gaacgctgaa aatccaggtc tctttgcaga 2040tcgggataaa gacagatgac agccacgatt ggaccaagct gcgctatatg gatagccata 2100cgccagcgga cgcggagcga gccggattgc ttgtaaggac ttcagcaccg tgcacgatca 2160ccgggaccat gggacacttt attctcgccc gatgcccgaa aggagagacg ctgacagtgg 2220gatttacgga cagcagaaag atcagccaca catgcacaca cccgttccat catgaaccac 2280ctgtgatagg tagggagagg ttccactctc gaccacaaca tggtaaagag ttaccttgca 2340gcacgtacgt gcagagcacc gctgccactg ctgaggagat agaggtgcat atgcccccag 2400atactcctga ccgcacgctg atgacgcagc agtctggcaa cgtgaagatc acagttaatg 2460ggcagacggt gcggtacaag tgcaactgcg gtggctccgg aggaaacccg aatgccaatc 2520ccaacgcgaa ccccaatgct aacccaaatg ccaacccaaa cgccaacccc aacgctggtg 2580gatccaacga gggactgaca accacagaca aagtgatcaa taactgcaaa attgatcagt 2640gccatgctgc agtcactaat cacaagaatt ggcaatacaa ctccccttta gtcccgcgca 2700acgctgaact cggggaccgt aaaggaaaga tccacatccc attcccattg gcaaacgtga 2760cttgcagagt gccaaaagca agaaacccta cagtaactta cggaaaaaac caagtcacca

2820tgctgctgta tcctgaccat ccgacactct tgtcttaccg taacatggga caggaaccaa 2880attaccacga ggagtgggtg acacacaaga aggaggttac cttgaccgtg cctactgagg 2940gtctggaggt cacttggggc aacaacgaac catacaagta ctggccgcag atgtctacga 3000acggtactgc tcatggtcac ccacatgaga taatcttgta ctattatgag ctgtacccca 3060ctatgactgt agtcattgtg tcggtggcct cgttcgtgct tctgtcgatg gtgggcacag 3120cagtgggaat gtgtgtgtgc gcacggcgca gatgcattac accatatgaa ttaacaccag 3180gagccactgt tcccttcctg ctcagcctgc tatgctgcgt cagaacgacc aaggcggcca 3240catattacga ggctgcggca tatctatgga acgaacagca gcccctgttc tggttgcagg 3300ctcttatccc gctggccgcc ttgatcgtcc tgtgcaactg tctgaaactc ttgccatgct 3360gctgtaagac cctggctttt ttagccgtaa tgagcatcgg tgcccacact gtgagcgcgt 3420acgaacacgt aacagtgatc ccgaacacgg tgggagtacc gtataagact cttgtcaaca 3480gaccgggtta cagccccatg gtgttggaga tggagctaca atcagtcacc ttggaaccaa 3540cactgtcact tgactacatc acgtgcgagt acaaaactgt catcccctcc ccgtacgtga 3600agtgctgtgg tacagcagag tgcaaggaca agagcctacc agactacagc tgcaaggtct 3660ttactggagt ctacccattt atgtggggcg gcgcctactg cttttgcgac gccgaaaata 3720cgcaattgag cgaggcacat gtagagaaat ctgaatcttg caaaacagag tttgcatcgg 3780cctacagagc ccacaccgca tcggcgtcgg cgaagctccg cgtcctttac caaggaaaca 3840acattaccgt agctgcctac gctaacggtg accatgccgt cacagtaaag gacgccaagt 3900ttgtcgtggg cccaatgtcc tccgcctgga caccttttga caacaaaatc gtggtgtaca 3960aaggcgacgt ctacaacatg gactacccac cttttggcgc aggaagacca ggacaatttg 4020gtgacattca aagtcgtaca ccggaaagta aagacgttta tgccaacact cagttggtac 4080tacagaggcc agcagcaggc acggtacatg taccatactc tcaggcacca tctggcttca 4140agtattggct gaaggaacga ggagcatcgc tacagcacac ggcaccgttc ggttgccaga 4200ttgcgacaaa cccggtaaga gctgtaaatt gcgctgtggg gaacatacca atttccatcg 4260acataccgga tgcggccttt actagggttg tcgatgcacc ctctgtaacg gacatgtcat 4320gcgaagtacc agcctgcact cactcctccg actttggggg cgtcgccatc atcaaataca 4380cagctagcaa gaaaggtaaa tgtgcagtac attcgatgac caacgccgtt accattcgag 4440aagccgacgt agaagtagag gggaactccc agctgcaaat atccttctca acagccctgg 4500caagcgccga gtttcgcgtg caagtgtgct ccacacaagt acactgcgca gccgcatgcc 4560accctccaaa ggaccacata gtcaattacc cagcatcaca caccaccctt ggggtccagg 4620atatatccac aacggcaatg tcttgggtgc agaagattac gggaggagta ggattaattg 4680ttgctgttgc tgccttaatt ttaattgtgg tgctatgcgt gtcgtttagc aggcactaag 4740gatctagatc tgctgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc 4800cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg 4860catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca 4920agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatgggta 4980cccaggtgct gaagaattga cccggttcct cctgggccag aaagaagcag gcacatcccc 5040ttctctgtga cacaccctgt ccacgcccct ggttcttagt tccagcccca ctcataggac 5100actcatagct caggagggct ccgccttcaa tcccacccgc taaagtactt ggagcggtct 5160ctccctccct catcagccca ccaaaccaaa cctagcctcc aagagtggga agaaattaaa 5220gcaagatagg ctattaagtg cagagggaga gaaaatgcct ccaacatgtg aggaagtaat 5280gagagaaatc atagaatttt aaggccatga tttaaggcca tcatggccta agcttgaaag 5340gagataggat caaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 5400tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 5460ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 5520aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 5580tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 5640gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 5700cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 5760gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 5820aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 5880ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 5940cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 6000ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 6060cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 6120agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 6180gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 6240gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 6300tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 6360agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 6420agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 6480atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 6540cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 6600actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 6660aatgataccg cgagaaccac gctcaccggc tccagattta tcagcaataa accagccagc 6720cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 6780ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 6840cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 6900ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 6960cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 7020ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 7080tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 7140ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 7200aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 7260gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 7320gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 7380ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 7440catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 7500atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 7560taaaaatagg cgtatcacga ggccctttcg ggtcgcgcgt ttcggtgatg acggtgaaaa 7620cctctgacac atgcagctcc cgttgacggt cacagcttgt ctgtaagcgg atgccgggag 7680cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 7740tgcggcatca gagcagattg tactgagagt gcaccataaa attgtaaacg ttaatatttt 7800gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat 7860cggcaaaatc ccttataaat caaaagaata gcccgagata gggttgagtg ttgttccagt 7920ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt 7980ctatcagggc gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag 8040gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg 8100aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc 8160gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc 8220gctacagggc gcgtactatg gttgctttga cgtatgcggt gtgaaatacc gcacagatgc 8280gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa 8340gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca 8400aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc 8460agtgaattcc atggtctcaa ctttc 8485273831DNAArtificial SequenceSynthetic VLP74_11 (CHI VLP 532 NPNAx6) CHIKV-NPNAx6 sequence 27atggagttca tcccgacgca aactttctat aacagaaggt accaaccccg accctgggcc 60ccacgcccta caattcaagt aattagacct agaccacgtc cacagaggca ggctgggcaa 120ctcgcccagc tgatctccgc agtcaacaaa ttgaccatgc gcgcggtacc tcaacagaag 180cctcgcagaa atcggaaaaa caagaagcaa aggcagaaga agcaggcgcc gcaaaacgac 240ccaaagcaaa agaagcaacc accacaaaag aagccggctc aaaagaagaa gaaaccaggc 300cgtagggaga gaatgtgcat gaaaattgaa aatgattgca tcttcgaagt caagcatgaa 360ggcaaagtga tgggctacgc atgcctggtg ggggataaag taatgaaacc agcacatgtg 420aagggaacta tcgacaatgc cgatctggct aaactggcct ttaagcggtc gtctaaatac 480gatcttgaat gtgcacagat accggtgcac atgaagtctg atgcctcgaa gtttacccac 540gagaaacccg aggggtacta taactggcat cacggagcag tgcagtattc aggaggccgg 600ttcactatcc cgacgggtgc aggcaagccg ggagacagcg gcagaccgat cttcgacaac 660aaaggacggg tggtggccat cgtcctagga ggggccaacg aaggtgcccg cacggccctc 720tccgtggtga cgtggaacaa agacatcgtc acaaaaatta cccctgaggg agccgaagag 780tggagcctcg ccctcccggt cttgtgcctg ttggcaaaca ctacattccc ctgctctcag 840ccgccttgca caccctgctg ctacgaaaag gaaccggaaa gcaccttgcg catgcttgag 900gacaacgtga tgagacccgg atactaccag ctactaaaag catcgctgac ttgctctccc 960caccgccaaa gacgcagtac taaggacaat tttaatgtct ataaagccac aagaccatat 1020ctagctcatt gtcctgactg cggagaaggg cattcgtgcc acagccctat cgcattggag 1080cgcatcagaa atgaagcaac ggacggaacg ctgaaaatcc aggtctcttt gcagatcggg 1140ataaagacag atgacagcca cgattggacc aagctgcgct atatggatag ccatacgcca 1200gcggacgcgg agcgagccgg attgcttgta aggacttcag caccgtgcac gatcaccggg 1260accatgggac actttattct cgcccgatgc ccgaaaggag agacgctgac agtgggattt 1320acggacagca gaaagatcag ccacacatgc acacacccgt tccatcatga accacctgtg 1380ataggtaggg agaggttcca ctctcgacca caacatggta aagagttacc ttgcagcacg 1440tacgtgcaga gcaccgctgc cactgctgag gagatagagg tgcatatgcc cccagatact 1500cctgaccgca cgctgatgac gcagcagtct ggcaacgtga agatcacagt taatgggcag 1560acggtgcggt acaagtgcaa ctgcggtggc tccggaggaa acccgaatgc caatcccaac 1620gcgaacccca atgctaaccc aaatgccaac ccaaacgcca accccaacgc tggtggatcc 1680aacgagggac tgacaaccac agacaaagtg atcaataact gcaaaattga tcagtgccat 1740gctgcagtca ctaatcacaa gaattggcaa tacaactccc ctttagtccc gcgcaacgct 1800gaactcgggg accgtaaagg aaagatccac atcccattcc cattggcaaa cgtgacttgc 1860agagtgccaa aagcaagaaa ccctacagta acttacggaa aaaaccaagt caccatgctg 1920ctgtatcctg accatccgac actcttgtct taccgtaaca tgggacagga accaaattac 1980cacgaggagt gggtgacaca caagaaggag gttaccttga ccgtgcctac tgagggtctg 2040gaggtcactt ggggcaacaa cgaaccatac aagtactggc cgcagatgtc tacgaacggt 2100actgctcatg gtcacccaca tgagataatc ttgtactatt atgagctgta ccccactatg 2160actgtagtca ttgtgtcggt ggcctcgttc gtgcttctgt cgatggtggg cacagcagtg 2220ggaatgtgtg tgtgcgcacg gcgcagatgc attacaccat atgaattaac accaggagcc 2280actgttccct tcctgctcag cctgctatgc tgcgtcagaa cgaccaaggc ggccacatat 2340tacgaggctg cggcatatct atggaacgaa cagcagcccc tgttctggtt gcaggctctt 2400atcccgctgg ccgccttgat cgtcctgtgc aactgtctga aactcttgcc atgctgctgt 2460aagaccctgg cttttttagc cgtaatgagc atcggtgccc acactgtgag cgcgtacgaa 2520cacgtaacag tgatcccgaa cacggtggga gtaccgtata agactcttgt caacagaccg 2580ggttacagcc ccatggtgtt ggagatggag ctacaatcag tcaccttgga accaacactg 2640tcacttgact acatcacgtg cgagtacaaa actgtcatcc cctccccgta cgtgaagtgc 2700tgtggtacag cagagtgcaa ggacaagagc ctaccagact acagctgcaa ggtctttact 2760ggagtctacc catttatgtg gggcggcgcc tactgctttt gcgacgccga aaatacgcaa 2820ttgagcgagg cacatgtaga gaaatctgaa tcttgcaaaa cagagtttgc atcggcctac 2880agagcccaca ccgcatcggc gtcggcgaag ctccgcgtcc tttaccaagg aaacaacatt 2940accgtagctg cctacgctaa cggtgaccat gccgtcacag taaaggacgc caagtttgtc 3000gtgggcccaa tgtcctccgc ctggacacct tttgacaaca aaatcgtggt gtacaaaggc 3060gacgtctaca acatggacta cccacctttt ggcgcaggaa gaccaggaca atttggtgac 3120attcaaagtc gtacaccgga aagtaaagac gtttatgcca acactcagtt ggtactacag 3180aggccagcag caggcacggt acatgtacca tactctcagg caccatctgg cttcaagtat 3240tggctgaagg aacgaggagc atcgctacag cacacggcac cgttcggttg ccagattgcg 3300acaaacccgg taagagctgt aaattgcgct gtggggaaca taccaatttc catcgacata 3360ccggatgcgg cctttactag ggttgtcgat gcaccctctg taacggacat gtcatgcgaa 3420gtaccagcct gcactcactc ctccgacttt gggggcgtcg ccatcatcaa atacacagct 3480agcaagaaag gtaaatgtgc agtacattcg atgaccaacg ccgttaccat tcgagaagcc 3540gacgtagaag tagaggggaa ctcccagctg caaatatcct tctcaacagc cctggcaagc 3600gccgagtttc gcgtgcaagt gtgctccaca caagtacact gcgcagccgc atgccaccct 3660ccaaaggacc acatagtcaa ttacccagca tcacacacca cccttggggt ccaggatata 3720tccacaacgg caatgtcttg ggtgcagaag attacgggag gagtaggatt aattgttgct 3780gttgctgcct taattttaat tgtggtgcta tgcgtgtcgt ttagcaggca c 3831281277PRTArtificial SequenceSynthetic VLP74_11 (CHI VLP 532 NPNAx6) sequence 28Met Glu Phe Ile Pro Thr Gln Thr Phe Tyr Asn Arg Arg Tyr Gln Pro 1 5 10 15 Arg Pro Trp Ala Pro Arg Pro Thr Ile Gln Val Ile Arg Pro Arg Pro 20 25 30 Arg Pro Gln Arg Gln Ala Gly Gln Leu Ala Gln Leu Ile Ser Ala Val 35 40 45 Asn Lys Leu Thr Met Arg Ala Val Pro Gln Gln Lys Pro Arg Arg Asn 50 55 60 Arg Lys Asn Lys Lys Gln Arg Gln Lys Lys Gln Ala Pro Gln Asn Asp 65 70 75 80 Pro Lys Gln Lys Lys Gln Pro Pro Gln Lys Lys Pro Ala Gln Lys Lys 85 90 95 Lys Lys Pro Gly Arg Arg Glu Arg Met Cys Met Lys Ile Glu Asn Asp 100 105 110 Cys Ile Phe Glu Val Lys His Glu Gly Lys Val Met Gly Tyr Ala Cys 115 120 125 Leu Val Gly Asp Lys Val Met Lys Pro Ala His Val Lys Gly Thr Ile 130 135 140 Asp Asn Ala Asp Leu Ala Lys Leu Ala Phe Lys Arg Ser Ser Lys Tyr 145 150 155 160 Asp Leu Glu Cys Ala Gln Ile Pro Val His Met Lys Ser Asp Ala Ser 165 170 175 Lys Phe Thr His Glu Lys Pro Glu Gly Tyr Tyr Asn Trp His His Gly 180 185 190 Ala Val Gln Tyr Ser Gly Gly Arg Phe Thr Ile Pro Thr Gly Ala Gly 195 200 205 Lys Pro Gly Asp Ser Gly Arg Pro Ile Phe Asp Asn Lys Gly Arg Val 210 215 220 Val Ala Ile Val Leu Gly Gly Ala Asn Glu Gly Ala Arg Thr Ala Leu 225 230 235 240 Ser Val Val Thr Trp Asn Lys Asp Ile Val Thr Lys Ile Thr Pro Glu 245 250 255 Gly Ala Glu Glu Trp Ser Leu Ala Leu Pro Val Leu Cys Leu Leu Ala 260 265 270 Asn Thr Thr Phe Pro Cys Ser Gln Pro Pro Cys Thr Pro Cys Cys Tyr 275 280 285 Glu Lys Glu Pro Glu Ser Thr Leu Arg Met Leu Glu Asp Asn Val Met 290 295 300 Arg Pro Gly Tyr Tyr Gln Leu Leu Lys Ala Ser Leu Thr Cys Ser Pro 305 310 315 320 His Arg Gln Arg Arg Ser Thr Lys Asp Asn Phe Asn Val Tyr Lys Ala 325 330 335 Thr Arg Pro Tyr Leu Ala His Cys Pro Asp Cys Gly Glu Gly His Ser 340 345 350 Cys His Ser Pro Ile Ala Leu Glu Arg Ile Arg Asn Glu Ala Thr Asp 355 360 365 Gly Thr Leu Lys Ile Gln Val Ser Leu Gln Ile Gly Ile Lys Thr Asp 370 375 380 Asp Ser His Asp Trp Thr Lys Leu Arg Tyr Met Asp Ser His Thr Pro 385 390 395 400 Ala Asp Ala Glu Arg Ala Gly Leu Leu Val Arg Thr Ser Ala Pro Cys 405 410 415 Thr Ile Thr Gly Thr Met Gly His Phe Ile Leu Ala Arg Cys Pro Lys 420 425 430 Gly Glu Thr Leu Thr Val Gly Phe Thr Asp Ser Arg Lys Ile Ser His 435 440 445 Thr Cys Thr His Pro Phe His His Glu Pro Pro Val Ile Gly Arg Glu 450 455 460 Arg Phe His Ser Arg Pro Gln His Gly Lys Glu Leu Pro Cys Ser Thr 465 470 475 480 Tyr Val Gln Ser Thr Ala Ala Thr Ala Glu Glu Ile Glu Val His Met 485 490 495 Pro Pro Asp Thr Pro Asp Arg Thr Leu Met Thr Gln Gln Ser Gly Asn 500 505 510 Val Lys Ile Thr Val Asn Gly Gln Thr Val Arg Tyr Lys Cys Asn Cys 515 520 525 Gly Gly Ser Gly Gly Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 530 535 540 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Gly Gly Ser 545 550 555 560 Asn Glu Gly Leu Thr Thr Thr Asp Lys Val Ile Asn Asn Cys Lys Ile 565 570 575 Asp Gln Cys His Ala Ala Val Thr Asn His Lys Asn Trp Gln Tyr Asn 580 585 590 Ser Pro Leu Val Pro Arg Asn Ala Glu Leu Gly Asp Arg Lys Gly Lys 595 600 605 Ile His Ile Pro Phe Pro Leu Ala Asn Val Thr Cys Arg Val Pro Lys 610 615 620 Ala Arg Asn Pro Thr Val Thr Tyr Gly Lys Asn Gln Val Thr Met Leu 625 630 635 640 Leu Tyr Pro Asp His Pro Thr Leu Leu Ser Tyr Arg Asn Met Gly Gln 645 650 655 Glu Pro Asn Tyr His Glu Glu Trp Val Thr His Lys Lys Glu Val Thr 660 665 670 Leu Thr Val Pro Thr Glu Gly Leu Glu Val Thr Trp Gly Asn Asn Glu 675 680 685 Pro Tyr Lys Tyr Trp Pro Gln Met Ser Thr Asn Gly Thr Ala His Gly 690 695 700 His Pro His Glu Ile Ile Leu Tyr Tyr Tyr Glu Leu Tyr Pro Thr Met 705 710 715 720 Thr Val Val Ile Val Ser Val Ala Ser Phe Val Leu Leu Ser Met Val 725 730 735 Gly Thr Ala Val Gly Met Cys Val Cys Ala Arg Arg Arg Cys Ile Thr 740 745 750 Pro Tyr Glu Leu Thr Pro Gly Ala Thr Val Pro Phe Leu Leu Ser Leu 755 760 765 Leu Cys Cys Val Arg Thr Thr Lys Ala Ala Thr Tyr Tyr Glu Ala Ala 770 775 780 Ala Tyr Leu Trp Asn Glu Gln Gln Pro Leu Phe Trp Leu Gln Ala Leu 785 790 795 800 Ile Pro Leu Ala Ala Leu Ile Val Leu Cys Asn Cys Leu Lys Leu Leu 805 810 815 Pro Cys Cys Cys Lys Thr Leu Ala Phe Leu

Ala Val Met Ser Ile Gly 820 825 830 Ala His Thr Val Ser Ala Tyr Glu His Val Thr Val Ile Pro Asn Thr 835 840 845 Val Gly Val Pro Tyr Lys Thr Leu Val Asn Arg Pro Gly Tyr Ser Pro 850 855 860 Met Val Leu Glu Met Glu Leu Gln Ser Val Thr Leu Glu Pro Thr Leu 865 870 875 880 Ser Leu Asp Tyr Ile Thr Cys Glu Tyr Lys Thr Val Ile Pro Ser Pro 885 890 895 Tyr Val Lys Cys Cys Gly Thr Ala Glu Cys Lys Asp Lys Ser Leu Pro 900 905 910 Asp Tyr Ser Cys Lys Val Phe Thr Gly Val Tyr Pro Phe Met Trp Gly 915 920 925 Gly Ala Tyr Cys Phe Cys Asp Ala Glu Asn Thr Gln Leu Ser Glu Ala 930 935 940 His Val Glu Lys Ser Glu Ser Cys Lys Thr Glu Phe Ala Ser Ala Tyr 945 950 955 960 Arg Ala His Thr Ala Ser Ala Ser Ala Lys Leu Arg Val Leu Tyr Gln 965 970 975 Gly Asn Asn Ile Thr Val Ala Ala Tyr Ala Asn Gly Asp His Ala Val 980 985 990 Thr Val Lys Asp Ala Lys Phe Val Val Gly Pro Met Ser Ser Ala Trp 995 1000 1005 Thr Pro Phe Asp Asn Lys Ile Val Val Tyr Lys Gly Asp Val Tyr 1010 1015 1020 Asn Met Asp Tyr Pro Pro Phe Gly Ala Gly Arg Pro Gly Gln Phe 1025 1030 1035 Gly Asp Ile Gln Ser Arg Thr Pro Glu Ser Lys Asp Val Tyr Ala 1040 1045 1050 Asn Thr Gln Leu Val Leu Gln Arg Pro Ala Ala Gly Thr Val His 1055 1060 1065 Val Pro Tyr Ser Gln Ala Pro Ser Gly Phe Lys Tyr Trp Leu Lys 1070 1075 1080 Glu Arg Gly Ala Ser Leu Gln His Thr Ala Pro Phe Gly Cys Gln 1085 1090 1095 Ile Ala Thr Asn Pro Val Arg Ala Val Asn Cys Ala Val Gly Asn 1100 1105 1110 Ile Pro Ile Ser Ile Asp Ile Pro Asp Ala Ala Phe Thr Arg Val 1115 1120 1125 Val Asp Ala Pro Ser Val Thr Asp Met Ser Cys Glu Val Pro Ala 1130 1135 1140 Cys Thr His Ser Ser Asp Phe Gly Gly Val Ala Ile Ile Lys Tyr 1145 1150 1155 Thr Ala Ser Lys Lys Gly Lys Cys Ala Val His Ser Met Thr Asn 1160 1165 1170 Ala Val Thr Ile Arg Glu Ala Asp Val Glu Val Glu Gly Asn Ser 1175 1180 1185 Gln Leu Gln Ile Ser Phe Ser Thr Ala Leu Ala Ser Ala Glu Phe 1190 1195 1200 Arg Val Gln Val Cys Ser Thr Gln Val His Cys Ala Ala Ala Cys 1205 1210 1215 His Pro Pro Lys Asp His Ile Val Asn Tyr Pro Ala Ser His Thr 1220 1225 1230 Thr Leu Gly Val Gln Asp Ile Ser Thr Thr Ala Met Ser Trp Val 1235 1240 1245 Gln Lys Ile Thr Gly Gly Val Gly Leu Ile Val Ala Val Ala Ala 1250 1255 1260 Leu Ile Leu Ile Val Val Leu Cys Val Ser Phe Ser Arg His 1265 1270 1275 298713DNAArtificial SequenceSynthetic VLP78_11 (CHI VLP 532 NPNAx25) sequence 29gaattcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt 60ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg 120gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 180ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 240atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 300gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 360gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 420tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 480atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 540gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 600tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga 660gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 720agaagacacc gggaccgatc cagcctccgt taacggtgga gggcagtgta gtctgagcag 780tactcgttgc tgccgcgcgc gccaccagac ataatagctg acagactaac agactgttcc 840tttccatggg tcttttctgc agtcaccgtc gtcgacacgt gtgatcagat atcgcggccg 900ccaccatgga gttcatcccg acgcaaactt tctataacag aaggtaccaa ccccgaccct 960gggccccacg ccctacaatt caagtaatta gacctagacc acgtccacag aggcaggctg 1020ggcaactcgc ccagctgatc tccgcagtca acaaattgac catgcgcgcg gtacctcaac 1080agaagcctcg cagaaatcgg aaaaacaaga agcaaaggca gaagaagcag gcgccgcaaa 1140acgacccaaa gcaaaagaag caaccaccac aaaagaagcc ggctcaaaag aagaagaaac 1200caggccgtag ggagagaatg tgcatgaaaa ttgaaaatga ttgcatcttc gaagtcaagc 1260atgaaggcaa agtgatgggc tacgcatgcc tggtggggga taaagtaatg aaaccagcac 1320atgtgaaggg aactatcgac aatgccgatc tggctaaact ggcctttaag cggtcgtcta 1380aatacgatct tgaatgtgca cagataccgg tgcacatgaa gtctgatgcc tcgaagttta 1440cccacgagaa acccgagggg tactataact ggcatcacgg agcagtgcag tattcaggag 1500gccggttcac tatcccgacg ggtgcaggca agccgggaga cagcggcaga ccgatcttcg 1560acaacaaagg acgggtggtg gccatcgtcc taggaggggc caacgaaggt gcccgcacgg 1620ccctctccgt ggtgacgtgg aacaaagaca tcgtcacaaa aattacccct gagggagccg 1680aagagtggag cctcgccctc ccggtcttgt gcctgttggc aaacactaca ttcccctgct 1740ctcagccgcc ttgcacaccc tgctgctacg aaaaggaacc ggaaagcacc ttgcgcatgc 1800ttgaggacaa cgtgatgaga cccggatact accagctact aaaagcatcg ctgacttgct 1860ctccccaccg ccaaagacgc agtactaagg acaattttaa tgtctataaa gccacaagac 1920catatctagc tcattgtcct gactgcggag aagggcattc gtgccacagc cctatcgcat 1980tggagcgcat cagaaatgaa gcaacggacg gaacgctgaa aatccaggtc tctttgcaga 2040tcgggataaa gacagatgac agccacgatt ggaccaagct gcgctatatg gatagccata 2100cgccagcgga cgcggagcga gccggattgc ttgtaaggac ttcagcaccg tgcacgatca 2160ccgggaccat gggacacttt attctcgccc gatgcccgaa aggagagacg ctgacagtgg 2220gatttacgga cagcagaaag atcagccaca catgcacaca cccgttccat catgaaccac 2280ctgtgatagg tagggagagg ttccactctc gaccacaaca tggtaaagag ttaccttgca 2340gcacgtacgt gcagagcacc gctgccactg ctgaggagat agaggtgcat atgcccccag 2400atactcctga ccgcacgctg atgacgcagc agtctggcaa cgtgaagatc acagttaatg 2460ggcagacggt gcggtacaag tgcaactgcg gtggctccgg aggaaacccg aatgccaatc 2520ccaacgcgaa ccccaacgct aaccccaacg ccaatccgaa tgcaaacccg aacgttgacc 2580caaacgccaa cccgaatgcc aatcccaacg cgaaccccaa tgctaaccca aatgccaacc 2640caaacgccaa ccccaacgct aatccaaacg ccaaccctaa cgccaatccc aacgcgaatc 2700ctaacgctaa tcccaacgca aatcccaatg ctaatccgaa cgcgaaccct aatgcaaacc 2760ccaacgccaa cccgaacgct aacccgaacg ctaatcccaa cgccggtgga tccaacgagg 2820gactgacaac cacagacaaa gtgatcaata actgcaaaat tgatcagtgc catgctgcag 2880tcactaatca caagaattgg caatacaact cccctttagt cccgcgcaac gctgaactcg 2940gggaccgtaa aggaaagatc cacatcccat tcccattggc aaacgtgact tgcagagtgc 3000caaaagcaag aaaccctaca gtaacttacg gaaaaaacca agtcaccatg ctgctgtatc 3060ctgaccatcc gacactcttg tcttaccgta acatgggaca ggaaccaaat taccacgagg 3120agtgggtgac acacaagaag gaggttacct tgaccgtgcc tactgagggt ctggaggtca 3180cttggggcaa caacgaacca tacaagtact ggccgcagat gtctacgaac ggtactgctc 3240atggtcaccc acatgagata atcttgtact attatgagct gtaccccact atgactgtag 3300tcattgtgtc ggtggcctcg ttcgtgcttc tgtcgatggt gggcacagca gtgggaatgt 3360gtgtgtgcgc acggcgcaga tgcattacac catatgaatt aacaccagga gccactgttc 3420ccttcctgct cagcctgcta tgctgcgtca gaacgaccaa ggcggccaca tattacgagg 3480ctgcggcata tctatggaac gaacagcagc ccctgttctg gttgcaggct cttatcccgc 3540tggccgcctt gatcgtcctg tgcaactgtc tgaaactctt gccatgctgc tgtaagaccc 3600tggctttttt agccgtaatg agcatcggtg cccacactgt gagcgcgtac gaacacgtaa 3660cagtgatccc gaacacggtg ggagtaccgt ataagactct tgtcaacaga ccgggttaca 3720gccccatggt gttggagatg gagctacaat cagtcacctt ggaaccaaca ctgtcacttg 3780actacatcac gtgcgagtac aaaactgtca tcccctcccc gtacgtgaag tgctgtggta 3840cagcagagtg caaggacaag agcctaccag actacagctg caaggtcttt actggagtct 3900acccatttat gtggggcggc gcctactgct tttgcgacgc cgaaaatacg caattgagcg 3960aggcacatgt agagaaatct gaatcttgca aaacagagtt tgcatcggcc tacagagccc 4020acaccgcatc ggcgtcggcg aagctccgcg tcctttacca aggaaacaac attaccgtag 4080ctgcctacgc taacggtgac catgccgtca cagtaaagga cgccaagttt gtcgtgggcc 4140caatgtcctc cgcctggaca ccttttgaca acaaaatcgt ggtgtacaaa ggcgacgtct 4200acaacatgga ctacccacct tttggcgcag gaagaccagg acaatttggt gacattcaaa 4260gtcgtacacc ggaaagtaaa gacgtttatg ccaacactca gttggtacta cagaggccag 4320cagcaggcac ggtacatgta ccatactctc aggcaccatc tggcttcaag tattggctga 4380aggaacgagg agcatcgcta cagcacacgg caccgttcgg ttgccagatt gcgacaaacc 4440cggtaagagc tgtaaattgc gctgtgggga acataccaat ttccatcgac ataccggatg 4500cggcctttac tagggttgtc gatgcaccct ctgtaacgga catgtcatgc gaagtaccag 4560cctgcactca ctcctccgac tttgggggcg tcgccatcat caaatacaca gctagcaaga 4620aaggtaaatg tgcagtacat tcgatgacca acgccgttac cattcgagaa gccgacgtag 4680aagtagaggg gaactcccag ctgcaaatat ccttctcaac agccctggca agcgccgagt 4740ttcgcgtgca agtgtgctcc acacaagtac actgcgcagc cgcatgccac cctccaaagg 4800accacatagt caattaccca gcatcacaca ccacccttgg ggtccaggat atatccacaa 4860cggcaatgtc ttgggtgcag aagattacgg gaggagtagg attaattgtt gctgttgctg 4920ccttaatttt aattgtggtg ctatgcgtgt cgtttagcag gcactaagga tctagatctg 4980ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc 5040tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc 5100tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt 5160gggaagacaa tagcaggcat gctggggatg cggtgggctc tatgggtacc caggtgctga 5220agaattgacc cggttcctcc tgggccagaa agaagcaggc acatcccctt ctctgtgaca 5280caccctgtcc acgcccctgg ttcttagttc cagccccact cataggacac tcatagctca 5340ggagggctcc gccttcaatc ccacccgcta aagtacttgg agcggtctct ccctccctca 5400tcagcccacc aaaccaaacc tagcctccaa gagtgggaag aaattaaagc aagataggct 5460attaagtgca gagggagaga aaatgcctcc aacatgtgag gaagtaatga gagaaatcat 5520agaattttaa ggccatgatt taaggccatc atggcctaag cttgaaagga gataggatca 5580aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 5640tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 5700ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 5760ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 5820ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 5880agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 5940catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6000tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6060gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6120ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6180cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6240caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 6300ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 6360taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 6420taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 6480cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 6540tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 6600gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 6660catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 6720atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 6780ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 6840gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 6900agaaccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 6960gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7020agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7080catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7140aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7200gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7260taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7320caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 7380ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7440ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 7500tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 7560aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 7620actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 7680catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 7740agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 7800tatcacgagg ccctttcggg tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 7860gcagctcccg ttgacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 7920tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 7980gcagattgta ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc 8040gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc 8100ttataaatca aaagaatagc ccgagatagg gttgagtgtt gttccagttt ggaacaagag 8160tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga 8220tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc 8280actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa 8340cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt 8400agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc 8460gtactatggt tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa 8520taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg 8580cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt 8640tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattccat 8700ggtctcaact ttc 8713304059DNAArtificial SequenceSynthetic VLP78_11 (CHI VLP 532 NPNAx25) CHIKV-NPNAx25 sequence 30atggagttca tcccgacgca aactttctat aacagaaggt accaaccccg accctgggcc 60ccacgcccta caattcaagt aattagacct agaccacgtc cacagaggca ggctgggcaa 120ctcgcccagc tgatctccgc agtcaacaaa ttgaccatgc gcgcggtacc tcaacagaag 180cctcgcagaa atcggaaaaa caagaagcaa aggcagaaga agcaggcgcc gcaaaacgac 240ccaaagcaaa agaagcaacc accacaaaag aagccggctc aaaagaagaa gaaaccaggc 300cgtagggaga gaatgtgcat gaaaattgaa aatgattgca tcttcgaagt caagcatgaa 360ggcaaagtga tgggctacgc atgcctggtg ggggataaag taatgaaacc agcacatgtg 420aagggaacta tcgacaatgc cgatctggct aaactggcct ttaagcggtc gtctaaatac 480gatcttgaat gtgcacagat accggtgcac atgaagtctg atgcctcgaa gtttacccac 540gagaaacccg aggggtacta taactggcat cacggagcag tgcagtattc aggaggccgg 600ttcactatcc cgacgggtgc aggcaagccg ggagacagcg gcagaccgat cttcgacaac 660aaaggacggg tggtggccat cgtcctagga ggggccaacg aaggtgcccg cacggccctc 720tccgtggtga cgtggaacaa agacatcgtc acaaaaatta cccctgaggg agccgaagag 780tggagcctcg ccctcccggt cttgtgcctg ttggcaaaca ctacattccc ctgctctcag 840ccgccttgca caccctgctg ctacgaaaag gaaccggaaa gcaccttgcg catgcttgag 900gacaacgtga tgagacccgg atactaccag ctactaaaag catcgctgac ttgctctccc 960caccgccaaa gacgcagtac taaggacaat tttaatgtct ataaagccac aagaccatat 1020ctagctcatt gtcctgactg cggagaaggg cattcgtgcc acagccctat cgcattggag 1080cgcatcagaa atgaagcaac ggacggaacg ctgaaaatcc aggtctcttt gcagatcggg 1140ataaagacag atgacagcca cgattggacc aagctgcgct atatggatag ccatacgcca 1200gcggacgcgg agcgagccgg attgcttgta aggacttcag caccgtgcac gatcaccggg 1260accatgggac actttattct cgcccgatgc ccgaaaggag agacgctgac agtgggattt 1320acggacagca gaaagatcag ccacacatgc acacacccgt tccatcatga accacctgtg 1380ataggtaggg agaggttcca ctctcgacca caacatggta aagagttacc ttgcagcacg 1440tacgtgcaga gcaccgctgc cactgctgag gagatagagg tgcatatgcc cccagatact 1500cctgaccgca cgctgatgac gcagcagtct ggcaacgtga agatcacagt taatgggcag 1560acggtgcggt acaagtgcaa ctgcggtggc tccggaggaa acccgaatgc caatcccaac 1620gcgaacccca acgctaaccc caacgccaat ccgaatgcaa acccgaacgt tgacccaaac 1680gccaacccga atgccaatcc caacgcgaac cccaatgcta acccaaatgc caacccaaac 1740gccaacccca acgctaatcc aaacgccaac cctaacgcca atcccaacgc gaatcctaac 1800gctaatccca acgcaaatcc caatgctaat ccgaacgcga accctaatgc aaaccccaac 1860gccaacccga acgctaaccc gaacgctaat cccaacgccg gtggatccaa cgagggactg 1920acaaccacag acaaagtgat caataactgc aaaattgatc agtgccatgc tgcagtcact 1980aatcacaaga attggcaata caactcccct ttagtcccgc gcaacgctga actcggggac 2040cgtaaaggaa agatccacat cccattccca ttggcaaacg tgacttgcag agtgccaaaa 2100gcaagaaacc ctacagtaac ttacggaaaa aaccaagtca ccatgctgct gtatcctgac 2160catccgacac tcttgtctta ccgtaacatg ggacaggaac caaattacca cgaggagtgg 2220gtgacacaca agaaggaggt taccttgacc gtgcctactg agggtctgga ggtcacttgg 2280ggcaacaacg aaccatacaa gtactggccg cagatgtcta cgaacggtac tgctcatggt 2340cacccacatg agataatctt gtactattat gagctgtacc ccactatgac tgtagtcatt 2400gtgtcggtgg cctcgttcgt gcttctgtcg atggtgggca cagcagtggg aatgtgtgtg 2460tgcgcacggc gcagatgcat tacaccatat gaattaacac caggagccac tgttcccttc 2520ctgctcagcc tgctatgctg cgtcagaacg accaaggcgg ccacatatta cgaggctgcg 2580gcatatctat ggaacgaaca gcagcccctg ttctggttgc aggctcttat cccgctggcc 2640gccttgatcg tcctgtgcaa ctgtctgaaa ctcttgccat gctgctgtaa gaccctggct 2700tttttagccg taatgagcat cggtgcccac actgtgagcg cgtacgaaca cgtaacagtg 2760atcccgaaca cggtgggagt accgtataag actcttgtca acagaccggg ttacagcccc 2820atggtgttgg agatggagct acaatcagtc accttggaac caacactgtc acttgactac 2880atcacgtgcg agtacaaaac tgtcatcccc tccccgtacg tgaagtgctg tggtacagca 2940gagtgcaagg acaagagcct accagactac agctgcaagg tctttactgg agtctaccca 3000tttatgtggg gcggcgccta ctgcttttgc gacgccgaaa atacgcaatt gagcgaggca 3060catgtagaga

aatctgaatc ttgcaaaaca gagtttgcat cggcctacag agcccacacc 3120gcatcggcgt cggcgaagct ccgcgtcctt taccaaggaa acaacattac cgtagctgcc 3180tacgctaacg gtgaccatgc cgtcacagta aaggacgcca agtttgtcgt gggcccaatg 3240tcctccgcct ggacaccttt tgacaacaaa atcgtggtgt acaaaggcga cgtctacaac 3300atggactacc caccttttgg cgcaggaaga ccaggacaat ttggtgacat tcaaagtcgt 3360acaccggaaa gtaaagacgt ttatgccaac actcagttgg tactacagag gccagcagca 3420ggcacggtac atgtaccata ctctcaggca ccatctggct tcaagtattg gctgaaggaa 3480cgaggagcat cgctacagca cacggcaccg ttcggttgcc agattgcgac aaacccggta 3540agagctgtaa attgcgctgt ggggaacata ccaatttcca tcgacatacc ggatgcggcc 3600tttactaggg ttgtcgatgc accctctgta acggacatgt catgcgaagt accagcctgc 3660actcactcct ccgactttgg gggcgtcgcc atcatcaaat acacagctag caagaaaggt 3720aaatgtgcag tacattcgat gaccaacgcc gttaccattc gagaagccga cgtagaagta 3780gaggggaact cccagctgca aatatccttc tcaacagccc tggcaagcgc cgagtttcgc 3840gtgcaagtgt gctccacaca agtacactgc gcagccgcat gccaccctcc aaaggaccac 3900atagtcaatt acccagcatc acacaccacc cttggggtcc aggatatatc cacaacggca 3960atgtcttggg tgcagaagat tacgggagga gtaggattaa ttgttgctgt tgctgcctta 4020attttaattg tggtgctatg cgtgtcgttt agcaggcac 4059311353PRTArtificial SequenceSynthetic VLP78_11 (CHI VLP 532 NPNAx25) sequence 31Met Glu Phe Ile Pro Thr Gln Thr Phe Tyr Asn Arg Arg Tyr Gln Pro 1 5 10 15 Arg Pro Trp Ala Pro Arg Pro Thr Ile Gln Val Ile Arg Pro Arg Pro 20 25 30 Arg Pro Gln Arg Gln Ala Gly Gln Leu Ala Gln Leu Ile Ser Ala Val 35 40 45 Asn Lys Leu Thr Met Arg Ala Val Pro Gln Gln Lys Pro Arg Arg Asn 50 55 60 Arg Lys Asn Lys Lys Gln Arg Gln Lys Lys Gln Ala Pro Gln Asn Asp 65 70 75 80 Pro Lys Gln Lys Lys Gln Pro Pro Gln Lys Lys Pro Ala Gln Lys Lys 85 90 95 Lys Lys Pro Gly Arg Arg Glu Arg Met Cys Met Lys Ile Glu Asn Asp 100 105 110 Cys Ile Phe Glu Val Lys His Glu Gly Lys Val Met Gly Tyr Ala Cys 115 120 125 Leu Val Gly Asp Lys Val Met Lys Pro Ala His Val Lys Gly Thr Ile 130 135 140 Asp Asn Ala Asp Leu Ala Lys Leu Ala Phe Lys Arg Ser Ser Lys Tyr 145 150 155 160 Asp Leu Glu Cys Ala Gln Ile Pro Val His Met Lys Ser Asp Ala Ser 165 170 175 Lys Phe Thr His Glu Lys Pro Glu Gly Tyr Tyr Asn Trp His His Gly 180 185 190 Ala Val Gln Tyr Ser Gly Gly Arg Phe Thr Ile Pro Thr Gly Ala Gly 195 200 205 Lys Pro Gly Asp Ser Gly Arg Pro Ile Phe Asp Asn Lys Gly Arg Val 210 215 220 Val Ala Ile Val Leu Gly Gly Ala Asn Glu Gly Ala Arg Thr Ala Leu 225 230 235 240 Ser Val Val Thr Trp Asn Lys Asp Ile Val Thr Lys Ile Thr Pro Glu 245 250 255 Gly Ala Glu Glu Trp Ser Leu Ala Leu Pro Val Leu Cys Leu Leu Ala 260 265 270 Asn Thr Thr Phe Pro Cys Ser Gln Pro Pro Cys Thr Pro Cys Cys Tyr 275 280 285 Glu Lys Glu Pro Glu Ser Thr Leu Arg Met Leu Glu Asp Asn Val Met 290 295 300 Arg Pro Gly Tyr Tyr Gln Leu Leu Lys Ala Ser Leu Thr Cys Ser Pro 305 310 315 320 His Arg Gln Arg Arg Ser Thr Lys Asp Asn Phe Asn Val Tyr Lys Ala 325 330 335 Thr Arg Pro Tyr Leu Ala His Cys Pro Asp Cys Gly Glu Gly His Ser 340 345 350 Cys His Ser Pro Ile Ala Leu Glu Arg Ile Arg Asn Glu Ala Thr Asp 355 360 365 Gly Thr Leu Lys Ile Gln Val Ser Leu Gln Ile Gly Ile Lys Thr Asp 370 375 380 Asp Ser His Asp Trp Thr Lys Leu Arg Tyr Met Asp Ser His Thr Pro 385 390 395 400 Ala Asp Ala Glu Arg Ala Gly Leu Leu Val Arg Thr Ser Ala Pro Cys 405 410 415 Thr Ile Thr Gly Thr Met Gly His Phe Ile Leu Ala Arg Cys Pro Lys 420 425 430 Gly Glu Thr Leu Thr Val Gly Phe Thr Asp Ser Arg Lys Ile Ser His 435 440 445 Thr Cys Thr His Pro Phe His His Glu Pro Pro Val Ile Gly Arg Glu 450 455 460 Arg Phe His Ser Arg Pro Gln His Gly Lys Glu Leu Pro Cys Ser Thr 465 470 475 480 Tyr Val Gln Ser Thr Ala Ala Thr Ala Glu Glu Ile Glu Val His Met 485 490 495 Pro Pro Asp Thr Pro Asp Arg Thr Leu Met Thr Gln Gln Ser Gly Asn 500 505 510 Val Lys Ile Thr Val Asn Gly Gln Thr Val Arg Tyr Lys Cys Asn Cys 515 520 525 Gly Gly Ser Gly Gly Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 530 535 540 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Val Asp Pro Asn 545 550 555 560 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 565 570 575 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 580 585 590 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 595 600 605 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 610 615 620 Ala Asn Pro Asn Ala Asn Pro Asn Ala Gly Gly Ser Asn Glu Gly Leu 625 630 635 640 Thr Thr Thr Asp Lys Val Ile Asn Asn Cys Lys Ile Asp Gln Cys His 645 650 655 Ala Ala Val Thr Asn His Lys Asn Trp Gln Tyr Asn Ser Pro Leu Val 660 665 670 Pro Arg Asn Ala Glu Leu Gly Asp Arg Lys Gly Lys Ile His Ile Pro 675 680 685 Phe Pro Leu Ala Asn Val Thr Cys Arg Val Pro Lys Ala Arg Asn Pro 690 695 700 Thr Val Thr Tyr Gly Lys Asn Gln Val Thr Met Leu Leu Tyr Pro Asp 705 710 715 720 His Pro Thr Leu Leu Ser Tyr Arg Asn Met Gly Gln Glu Pro Asn Tyr 725 730 735 His Glu Glu Trp Val Thr His Lys Lys Glu Val Thr Leu Thr Val Pro 740 745 750 Thr Glu Gly Leu Glu Val Thr Trp Gly Asn Asn Glu Pro Tyr Lys Tyr 755 760 765 Trp Pro Gln Met Ser Thr Asn Gly Thr Ala His Gly His Pro His Glu 770 775 780 Ile Ile Leu Tyr Tyr Tyr Glu Leu Tyr Pro Thr Met Thr Val Val Ile 785 790 795 800 Val Ser Val Ala Ser Phe Val Leu Leu Ser Met Val Gly Thr Ala Val 805 810 815 Gly Met Cys Val Cys Ala Arg Arg Arg Cys Ile Thr Pro Tyr Glu Leu 820 825 830 Thr Pro Gly Ala Thr Val Pro Phe Leu Leu Ser Leu Leu Cys Cys Val 835 840 845 Arg Thr Thr Lys Ala Ala Thr Tyr Tyr Glu Ala Ala Ala Tyr Leu Trp 850 855 860 Asn Glu Gln Gln Pro Leu Phe Trp Leu Gln Ala Leu Ile Pro Leu Ala 865 870 875 880 Ala Leu Ile Val Leu Cys Asn Cys Leu Lys Leu Leu Pro Cys Cys Cys 885 890 895 Lys Thr Leu Ala Phe Leu Ala Val Met Ser Ile Gly Ala His Thr Val 900 905 910 Ser Ala Tyr Glu His Val Thr Val Ile Pro Asn Thr Val Gly Val Pro 915 920 925 Tyr Lys Thr Leu Val Asn Arg Pro Gly Tyr Ser Pro Met Val Leu Glu 930 935 940 Met Glu Leu Gln Ser Val Thr Leu Glu Pro Thr Leu Ser Leu Asp Tyr 945 950 955 960 Ile Thr Cys Glu Tyr Lys Thr Val Ile Pro Ser Pro Tyr Val Lys Cys 965 970 975 Cys Gly Thr Ala Glu Cys Lys Asp Lys Ser Leu Pro Asp Tyr Ser Cys 980 985 990 Lys Val Phe Thr Gly Val Tyr Pro Phe Met Trp Gly Gly Ala Tyr Cys 995 1000 1005 Phe Cys Asp Ala Glu Asn Thr Gln Leu Ser Glu Ala His Val Glu 1010 1015 1020 Lys Ser Glu Ser Cys Lys Thr Glu Phe Ala Ser Ala Tyr Arg Ala 1025 1030 1035 His Thr Ala Ser Ala Ser Ala Lys Leu Arg Val Leu Tyr Gln Gly 1040 1045 1050 Asn Asn Ile Thr Val Ala Ala Tyr Ala Asn Gly Asp His Ala Val 1055 1060 1065 Thr Val Lys Asp Ala Lys Phe Val Val Gly Pro Met Ser Ser Ala 1070 1075 1080 Trp Thr Pro Phe Asp Asn Lys Ile Val Val Tyr Lys Gly Asp Val 1085 1090 1095 Tyr Asn Met Asp Tyr Pro Pro Phe Gly Ala Gly Arg Pro Gly Gln 1100 1105 1110 Phe Gly Asp Ile Gln Ser Arg Thr Pro Glu Ser Lys Asp Val Tyr 1115 1120 1125 Ala Asn Thr Gln Leu Val Leu Gln Arg Pro Ala Ala Gly Thr Val 1130 1135 1140 His Val Pro Tyr Ser Gln Ala Pro Ser Gly Phe Lys Tyr Trp Leu 1145 1150 1155 Lys Glu Arg Gly Ala Ser Leu Gln His Thr Ala Pro Phe Gly Cys 1160 1165 1170 Gln Ile Ala Thr Asn Pro Val Arg Ala Val Asn Cys Ala Val Gly 1175 1180 1185 Asn Ile Pro Ile Ser Ile Asp Ile Pro Asp Ala Ala Phe Thr Arg 1190 1195 1200 Val Val Asp Ala Pro Ser Val Thr Asp Met Ser Cys Glu Val Pro 1205 1210 1215 Ala Cys Thr His Ser Ser Asp Phe Gly Gly Val Ala Ile Ile Lys 1220 1225 1230 Tyr Thr Ala Ser Lys Lys Gly Lys Cys Ala Val His Ser Met Thr 1235 1240 1245 Asn Ala Val Thr Ile Arg Glu Ala Asp Val Glu Val Glu Gly Asn 1250 1255 1260 Ser Gln Leu Gln Ile Ser Phe Ser Thr Ala Leu Ala Ser Ala Glu 1265 1270 1275 Phe Arg Val Gln Val Cys Ser Thr Gln Val His Cys Ala Ala Ala 1280 1285 1290 Cys His Pro Pro Lys Asp His Ile Val Asn Tyr Pro Ala Ser His 1295 1300 1305 Thr Thr Leu Gly Val Gln Asp Ile Ser Thr Thr Ala Met Ser Trp 1310 1315 1320 Val Gln Lys Ile Thr Gly Gly Val Gly Leu Ile Val Ala Val Ala 1325 1330 1335 Ala Leu Ile Leu Ile Val Val Leu Cys Val Ser Phe Ser Arg His 1340 1345 1350 328737DNAArtificial SequenceSynthetic VLP78_21 (VEEV VLP 519 NPNAx25) sequence 32gaattcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt 60ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg 120gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 180ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 240atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 300gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 360gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 420tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 480atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 540gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 600tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga 660gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 720agaagacacc gggaccgatc cagcctccgt taacggtgga gggcagtgta gtctgagcag 780tactcgttgc tgccgcgcgc gccaccagac ataatagctg acagactaac agactgttcc 840tttccatggg tcttttctgc agtcaccgtc gtcgacacgt gtgatcagat atcgcggccg 900ccaccatgtt cccgttccag ccaatgtatc cgatgcagcc aatgccctat cgcaacccgt 960tcgcggcccc gcgcaggccc tggttcccca gaaccgaccc ttttctggcg atgcaggtgc 1020aggaattaac ccgctcgatg gctaacctga cgttcaagca acgccgggac gcgccacctg 1080aggggccatc cgctaataaa ccgaagaagg aggcctcgca aaaacagaaa gggggaggcc 1140aagggaagaa gaagaagaac caagggaaga agaaggctaa gacagggccg cctaatccga 1200aggcacagaa tggaaacaag aagaagacca acaagaaacc aggcaagaga cagcgcatgg 1260tcatgaaatt ggaatctgac aagacgttcc caatcatgtt ggaagggaag ataaacggct 1320acgcttgtgt ggtcggaggg aagttattca ggccgatgca tgtggaaggc aagatcgaca 1380acgacgttct ggccgcgctt aagacgaaga aagcatccaa atacgatctt gagtatgcag 1440atgtgccaca gaacatgcgg gccgatacat tcaaatacac ccatgagaaa ccccaaggct 1500attacagctg gcatcatgga gcagtccaat atgaaaatgg gcgtttcacg gtgccgaaag 1560gagttggggc caagggagac agcggacgac ccattctgga taaccaggga cgggtggtcg 1620ctattgtgct gggaggtgtg aatgaaggat ctaggacagc cctttcagtc gtcatgtgga 1680acgagaaggg agttaccgtg aagtatactc cagagaactg cgagcaatgg tcactagtga 1740ccaccatgtg tctgctcgcc aatgtgacgt tcccatgtgc tcaaccacca atttgctacg 1800acagaaaacc agcagagact ttggccatgc tcagcgttaa cgttgacaac ccgggctacg 1860atgagctgct ggaagcagct gttaagtgcc ccggaaggaa aaggagatcc accgaggagc 1920tgtttaatga gtataagcta acgcgccctt acatggccag atgcatcaga tgtgcagttg 1980ggagctgcca tagtccaata gcaatcgagg cagtaaagag cgacgggcac gacggttatg 2040ttagacttca gacttcctcg cagtatggcc tggattcctc cggcaactta aagggcagga 2100ccatgcggta tgacatgcac gggaccatta aagagatacc actacatcaa gtgtcactct 2160atacatctcg cccgtgtcac attgtggatg ggcacggtta tttcctgctt gccaggtgcc 2220cggcagggga ctccatcacc atggaattta agaaagattc cgtcagacac tcctgctcgg 2280tgccgtatga agtgaaattt aatcctgtag gcagagaact ctatactcat cccccagaac 2340acggagtaga gcaagcgtgc caagtctacg cacatgatgc acagaacaga ggagcttatg 2400tcgagatgca cctcccgggc tcagaagtgg acagcagttt ggtttccttg agcggcagtt 2460ccggaggaaa cccgaatgcc aatcccaacg cgaaccccaa cgctaacccc aacgccaatc 2520cgaatgcaaa cccgaacgtt gacccaaacg ccaacccgaa tgccaatccc aacgcgaacc 2580ccaatgctaa cccaaatgcc aacccaaacg ccaaccccaa cgctaatcca aacgccaacc 2640ctaacgccaa tcccaacgcg aatcctaacg ctaatcccaa cgcaaatccc aatgctaatc 2700cgaacgcgaa ccctaatgca aaccccaacg ccaacccgaa cgctaacccg aacgctaatc 2760ccaacgccgg tggatcctca gtcaccgtga cacctcctga tgggactagc gccctggtgg 2820aatgcgagtg tggcggcaca aagatctccg agaccatcaa caagacaaaa cagttcagcc 2880agtgcacaaa gaaggagcag tgcagagcat atcggctgca gaacgataag tgggtgtata 2940attctgacaa actgcccaaa gcagcgggag ccaccttaaa aggaaaactg catgtcccat 3000tcttgctggc agacggcaaa tgcaccgtgc ctctagcacc agaacctatg ataaccttcg 3060gtttcagatc agtgtcactg aaactgcacc ctaagaatcc cacatatcta atcacccgcc 3120aacttgctga tgagcctcac tacacgcacg agctcatatc tgaaccagct gttaggaatt 3180ttaccgtcac cgaaaaaggg tgggagtttg tatggggaaa ccacccgccg aaaaggtttt 3240gggcacagga aacagcaccc ggaaatccac atgggctacc gcacgaggtg ataactcatt 3300attaccacag ataccctatg tccaccatcc tgggtttgtc aatttgtgcc gccattgcaa 3360ccgtttccgt tgcagcgtct acctggctgt tttgcagatc aagagttgcg tgcctaactc 3420cttaccggct aacacctaac gctaggatac cattttgtct ggctgtgctt tgctgcgccc 3480gcactgcccg ggccgagacc acctgggagt ccttggatca cctatggaac aataaccaac 3540agatgttctg gattcaattg ctgatccctc tggccgcctt gatcgtagtg actcgcctgc 3600tcaggtgcgt gtgctgtgtc gtgccttttt tagtcatggc cggcgccgca ggcgccggcg 3660cctacgagca cgcgaccacg atgccgagcc aagcgggaat ctcgtataac actatagtca 3720acagagcagg ctacgcacca ctccctatca gcataacacc aacaaagatc aagctgatac 3780ctacagtgaa cttggagtac gtcacctgcc actacaaaac aggaatggat tcaccagcca 3840tcaaatgctg cggatctcag gaatgcactc caacttacag gcctgatgaa cagtgcaaag 3900tcttcacagg ggtttacccg ttcatgtggg gtggtgcata ttgcttttgc gacactgaga 3960acacccaagt cagcaaggcc tacgtaatga aatctgacga ctgccttgcg gatcatgctg 4020aagcatataa agcgcacaca gcctcagtgc aggcgttcct caacatcaca gtgggagaac 4080actctattgt gactaccgtg tatgtgaatg gagaaactcc tgtgaatttc aatggggtca 4140aaataactgc aggtccgctt tccacagctt ggacaccctt tgatcgcaaa atcgtgcagt 4200atgccgggga gatctataat tatgattttc ctgagtatgg ggcaggacaa ccaggagcat 4260ttggagatat acaatccaga acagtctcaa gctctgatct gtatgccaat accaacctag 4320tgctgcagag acccaaagca ggagcgatcc acgtgccata cactcaggca ccttcgggtt 4380ttgagcaatg gaagaaagat aaagctccat cattgaaatt taccgcccct ttcggatgcg 4440aaatatatac aaaccccatt cgcgccgaaa actgtgctgt agggtcaatt ccattagcct 4500ttgacattcc cgacgccttg ttcaccaggg tgtcagaaac accgacactt tcagcggccg 4560aatgcactct taacgagtgc gtgtattctt ccgactttgg tgggatcgcc acggtcaagt 4620actcggccag caagtcaggc aagtgcgcag tccatgtgcc atcagggact gctaccctaa 4680aagaagcagc agtcgagcta accgagcaag ggtcggcgac tatccatttc tcgaccgcaa 4740atatccaccc ggagttcagg ctccaaatat gcacatcata tgttacgtgc aaaggtgatt 4800gtcacccccc gaaagaccat attgtgacac accctcagta tcacgcccaa acatttacag 4860ccgcggtgtc aaaaaccgcg tggacgtggt taacatccct gctgggagga tcagccgtaa 4920ttattataat tggcttggtg

ctggctacta ttgtggccat gtacgtgctg accaaccaga 4980aacataatta aggatctaga tctgctgtgc cttctagttg ccagccatct gttgtttgcc 5040cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 5100atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 5160ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 5220gctctatggg tacccaggtg ctgaagaatt gacccggttc ctcctgggcc agaaagaagc 5280aggcacatcc ccttctctgt gacacaccct gtccacgccc ctggttctta gttccagccc 5340cactcatagg acactcatag ctcaggaggg ctccgccttc aatcccaccc gctaaagtac 5400ttggagcggt ctctccctcc ctcatcagcc caccaaacca aacctagcct ccaagagtgg 5460gaagaaatta aagcaagata ggctattaag tgcagaggga gagaaaatgc ctccaacatg 5520tgaggaagta atgagagaaa tcatagaatt ttaaggccat gatttaaggc catcatggcc 5580taagcttgaa aggagatagg atcaaagctt ggcgtaatca tggtcatagc tgtttcctgt 5640gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 5700agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 5760tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 5820aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 5880cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 5940atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 6000taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 6060aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 6120tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 6180gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 6240cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 6300cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 6360atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 6420tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 6480ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 6540acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 6600aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 6660aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 6720tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 6780cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 6840catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 6900ccccagtgct gcaatgatac cgcgagaacc acgctcaccg gctccagatt tatcagcaat 6960aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 7020ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 7080caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 7140attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 7200agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 7260actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 7320ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 7380ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 7440gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 7500atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 7560cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 7620gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 7680gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 7740ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat 7800gacattaacc tataaaaata ggcgtatcac gaggcccttt cgggtcgcgc gtttcggtga 7860tgacggtgaa aacctctgac acatgcagct cccgttgacg gtcacagctt gtctgtaagc 7920ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg 7980ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata aaattgtaaa 8040cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca 8100ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga tagggttgag 8160tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg 8220gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt 8280tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag 8340agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc 8400gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc 8460gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt gacgtatgcg gtgtgaaata 8520ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc 8580aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg 8640ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt 8700aaaacgacgg ccagtgaatt ccatggtctc aactttc 8737334086DNAArtificial SequenceSynthetic VLP78_21 (VEEV VLP 519 NPNAx25) VEEV-NPNAx25 sequence 33atgttcccgt tccagccaat gtatccgatg cagccaatgc cctatcgcaa cccgttcgcg 60gccccgcgca ggccctggtt ccccagaacc gacccttttc tggcgatgca ggtgcaggaa 120ttaacccgct cgatggctaa cctgacgttc aagcaacgcc gggacgcgcc acctgagggg 180ccatccgcta ataaaccgaa gaaggaggcc tcgcaaaaac agaaaggggg aggccaaggg 240aagaagaaga agaaccaagg gaagaagaag gctaagacag ggccgcctaa tccgaaggca 300cagaatggaa acaagaagaa gaccaacaag aaaccaggca agagacagcg catggtcatg 360aaattggaat ctgacaagac gttcccaatc atgttggaag ggaagataaa cggctacgct 420tgtgtggtcg gagggaagtt attcaggccg atgcatgtgg aaggcaagat cgacaacgac 480gttctggccg cgcttaagac gaagaaagca tccaaatacg atcttgagta tgcagatgtg 540ccacagaaca tgcgggccga tacattcaaa tacacccatg agaaacccca aggctattac 600agctggcatc atggagcagt ccaatatgaa aatgggcgtt tcacggtgcc gaaaggagtt 660ggggccaagg gagacagcgg acgacccatt ctggataacc agggacgggt ggtcgctatt 720gtgctgggag gtgtgaatga aggatctagg acagcccttt cagtcgtcat gtggaacgag 780aagggagtta ccgtgaagta tactccagag aactgcgagc aatggtcact agtgaccacc 840atgtgtctgc tcgccaatgt gacgttccca tgtgctcaac caccaatttg ctacgacaga 900aaaccagcag agactttggc catgctcagc gttaacgttg acaacccggg ctacgatgag 960ctgctggaag cagctgttaa gtgccccgga aggaaaagga gatccaccga ggagctgttt 1020aatgagtata agctaacgcg cccttacatg gccagatgca tcagatgtgc agttgggagc 1080tgccatagtc caatagcaat cgaggcagta aagagcgacg ggcacgacgg ttatgttaga 1140cttcagactt cctcgcagta tggcctggat tcctccggca acttaaaggg caggaccatg 1200cggtatgaca tgcacgggac cattaaagag ataccactac atcaagtgtc actctataca 1260tctcgcccgt gtcacattgt ggatgggcac ggttatttcc tgcttgccag gtgcccggca 1320ggggactcca tcaccatgga atttaagaaa gattccgtca gacactcctg ctcggtgccg 1380tatgaagtga aatttaatcc tgtaggcaga gaactctata ctcatccccc agaacacgga 1440gtagagcaag cgtgccaagt ctacgcacat gatgcacaga acagaggagc ttatgtcgag 1500atgcacctcc cgggctcaga agtggacagc agtttggttt ccttgagcgg cagttccgga 1560ggaaacccga atgccaatcc caacgcgaac cccaacgcta accccaacgc caatccgaat 1620gcaaacccga acgttgaccc aaacgccaac ccgaatgcca atcccaacgc gaaccccaat 1680gctaacccaa atgccaaccc aaacgccaac cccaacgcta atccaaacgc caaccctaac 1740gccaatccca acgcgaatcc taacgctaat cccaacgcaa atcccaatgc taatccgaac 1800gcgaacccta atgcaaaccc caacgccaac ccgaacgcta acccgaacgc taatcccaac 1860gccggtggat cctcagtcac cgtgacacct cctgatggga ctagcgccct ggtggaatgc 1920gagtgtggcg gcacaaagat ctccgagacc atcaacaaga caaaacagtt cagccagtgc 1980acaaagaagg agcagtgcag agcatatcgg ctgcagaacg ataagtgggt gtataattct 2040gacaaactgc ccaaagcagc gggagccacc ttaaaaggaa aactgcatgt cccattcttg 2100ctggcagacg gcaaatgcac cgtgcctcta gcaccagaac ctatgataac cttcggtttc 2160agatcagtgt cactgaaact gcaccctaag aatcccacat atctaatcac ccgccaactt 2220gctgatgagc ctcactacac gcacgagctc atatctgaac cagctgttag gaattttacc 2280gtcaccgaaa aagggtggga gtttgtatgg ggaaaccacc cgccgaaaag gttttgggca 2340caggaaacag cacccggaaa tccacatggg ctaccgcacg aggtgataac tcattattac 2400cacagatacc ctatgtccac catcctgggt ttgtcaattt gtgccgccat tgcaaccgtt 2460tccgttgcag cgtctacctg gctgttttgc agatcaagag ttgcgtgcct aactccttac 2520cggctaacac ctaacgctag gataccattt tgtctggctg tgctttgctg cgcccgcact 2580gcccgggccg agaccacctg ggagtccttg gatcacctat ggaacaataa ccaacagatg 2640ttctggattc aattgctgat ccctctggcc gccttgatcg tagtgactcg cctgctcagg 2700tgcgtgtgct gtgtcgtgcc ttttttagtc atggccggcg ccgcaggcgc cggcgcctac 2760gagcacgcga ccacgatgcc gagccaagcg ggaatctcgt ataacactat agtcaacaga 2820gcaggctacg caccactccc tatcagcata acaccaacaa agatcaagct gatacctaca 2880gtgaacttgg agtacgtcac ctgccactac aaaacaggaa tggattcacc agccatcaaa 2940tgctgcggat ctcaggaatg cactccaact tacaggcctg atgaacagtg caaagtcttc 3000acaggggttt acccgttcat gtggggtggt gcatattgct tttgcgacac tgagaacacc 3060caagtcagca aggcctacgt aatgaaatct gacgactgcc ttgcggatca tgctgaagca 3120tataaagcgc acacagcctc agtgcaggcg ttcctcaaca tcacagtggg agaacactct 3180attgtgacta ccgtgtatgt gaatggagaa actcctgtga atttcaatgg ggtcaaaata 3240actgcaggtc cgctttccac agcttggaca ccctttgatc gcaaaatcgt gcagtatgcc 3300ggggagatct ataattatga ttttcctgag tatggggcag gacaaccagg agcatttgga 3360gatatacaat ccagaacagt ctcaagctct gatctgtatg ccaataccaa cctagtgctg 3420cagagaccca aagcaggagc gatccacgtg ccatacactc aggcaccttc gggttttgag 3480caatggaaga aagataaagc tccatcattg aaatttaccg cccctttcgg atgcgaaata 3540tatacaaacc ccattcgcgc cgaaaactgt gctgtagggt caattccatt agcctttgac 3600attcccgacg ccttgttcac cagggtgtca gaaacaccga cactttcagc ggccgaatgc 3660actcttaacg agtgcgtgta ttcttccgac tttggtggga tcgccacggt caagtactcg 3720gccagcaagt caggcaagtg cgcagtccat gtgccatcag ggactgctac cctaaaagaa 3780gcagcagtcg agctaaccga gcaagggtcg gcgactatcc atttctcgac cgcaaatatc 3840cacccggagt tcaggctcca aatatgcaca tcatatgtta cgtgcaaagg tgattgtcac 3900cccccgaaag accatattgt gacacaccct cagtatcacg cccaaacatt tacagccgcg 3960gtgtcaaaaa ccgcgtggac gtggttaaca tccctgctgg gaggatcagc cgtaattatt 4020ataattggct tggtgctggc tactattgtg gccatgtacg tgctgaccaa ccagaaacat 4080aattaa 4086341361PRTArtificial SequenceSynthetic VLP78_21 (VEEV VLP 519 NPNAx25) sequence 34Met Phe Pro Phe Gln Pro Met Tyr Pro Met Gln Pro Met Pro Tyr Arg 1 5 10 15 Asn Pro Phe Ala Ala Pro Arg Arg Pro Trp Phe Pro Arg Thr Asp Pro 20 25 30 Phe Leu Ala Met Gln Val Gln Glu Leu Thr Arg Ser Met Ala Asn Leu 35 40 45 Thr Phe Lys Gln Arg Arg Asp Ala Pro Pro Glu Gly Pro Ser Ala Asn 50 55 60 Lys Pro Lys Lys Glu Ala Ser Gln Lys Gln Lys Gly Gly Gly Gln Gly 65 70 75 80 Lys Lys Lys Lys Asn Gln Gly Lys Lys Lys Ala Lys Thr Gly Pro Pro 85 90 95 Asn Pro Lys Ala Gln Asn Gly Asn Lys Lys Lys Thr Asn Lys Lys Pro 100 105 110 Gly Lys Arg Gln Arg Met Val Met Lys Leu Glu Ser Asp Lys Thr Phe 115 120 125 Pro Ile Met Leu Glu Gly Lys Ile Asn Gly Tyr Ala Cys Val Val Gly 130 135 140 Gly Lys Leu Phe Arg Pro Met His Val Glu Gly Lys Ile Asp Asn Asp 145 150 155 160 Val Leu Ala Ala Leu Lys Thr Lys Lys Ala Ser Lys Tyr Asp Leu Glu 165 170 175 Tyr Ala Asp Val Pro Gln Asn Met Arg Ala Asp Thr Phe Lys Tyr Thr 180 185 190 His Glu Lys Pro Gln Gly Tyr Tyr Ser Trp His His Gly Ala Val Gln 195 200 205 Tyr Glu Asn Gly Arg Phe Thr Val Pro Lys Gly Val Gly Ala Lys Gly 210 215 220 Asp Ser Gly Arg Pro Ile Leu Asp Asn Gln Gly Arg Val Val Ala Ile 225 230 235 240 Val Leu Gly Gly Val Asn Glu Gly Ser Arg Thr Ala Leu Ser Val Val 245 250 255 Met Trp Asn Glu Lys Gly Val Thr Val Lys Tyr Thr Pro Glu Asn Cys 260 265 270 Glu Gln Trp Ser Leu Val Thr Thr Met Cys Leu Leu Ala Asn Val Thr 275 280 285 Phe Pro Cys Ala Gln Pro Pro Ile Cys Tyr Asp Arg Lys Pro Ala Glu 290 295 300 Thr Leu Ala Met Leu Ser Val Asn Val Asp Asn Pro Gly Tyr Asp Glu 305 310 315 320 Leu Leu Glu Ala Ala Val Lys Cys Pro Gly Arg Lys Arg Arg Ser Thr 325 330 335 Glu Glu Leu Phe Asn Glu Tyr Lys Leu Thr Arg Pro Tyr Met Ala Arg 340 345 350 Cys Ile Arg Cys Ala Val Gly Ser Cys His Ser Pro Ile Ala Ile Glu 355 360 365 Ala Val Lys Ser Asp Gly His Asp Gly Tyr Val Arg Leu Gln Thr Ser 370 375 380 Ser Gln Tyr Gly Leu Asp Ser Ser Gly Asn Leu Lys Gly Arg Thr Met 385 390 395 400 Arg Tyr Asp Met His Gly Thr Ile Lys Glu Ile Pro Leu His Gln Val 405 410 415 Ser Leu Tyr Thr Ser Arg Pro Cys His Ile Val Asp Gly His Gly Tyr 420 425 430 Phe Leu Leu Ala Arg Cys Pro Ala Gly Asp Ser Ile Thr Met Glu Phe 435 440 445 Lys Lys Asp Ser Val Arg His Ser Cys Ser Val Pro Tyr Glu Val Lys 450 455 460 Phe Asn Pro Val Gly Arg Glu Leu Tyr Thr His Pro Pro Glu His Gly 465 470 475 480 Val Glu Gln Ala Cys Gln Val Tyr Ala His Asp Ala Gln Asn Arg Gly 485 490 495 Ala Tyr Val Glu Met His Leu Pro Gly Ser Glu Val Asp Ser Ser Leu 500 505 510 Val Ser Leu Ser Gly Ser Ser Gly Gly Asn Pro Asn Ala Asn Pro Asn 515 520 525 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 530 535 540 Val Asp Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 545 550 555 560 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 565 570 575 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 580 585 590 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 595 600 605 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Gly Gly Ser 610 615 620 Ser Val Thr Val Thr Pro Pro Asp Gly Thr Ser Ala Leu Val Glu Cys 625 630 635 640 Glu Cys Gly Gly Thr Lys Ile Ser Glu Thr Ile Asn Lys Thr Lys Gln 645 650 655 Phe Ser Gln Cys Thr Lys Lys Glu Gln Cys Arg Ala Tyr Arg Leu Gln 660 665 670 Asn Asp Lys Trp Val Tyr Asn Ser Asp Lys Leu Pro Lys Ala Ala Gly 675 680 685 Ala Thr Leu Lys Gly Lys Leu His Val Pro Phe Leu Leu Ala Asp Gly 690 695 700 Lys Cys Thr Val Pro Leu Ala Pro Glu Pro Met Ile Thr Phe Gly Phe 705 710 715 720 Arg Ser Val Ser Leu Lys Leu His Pro Lys Asn Pro Thr Tyr Leu Ile 725 730 735 Thr Arg Gln Leu Ala Asp Glu Pro His Tyr Thr His Glu Leu Ile Ser 740 745 750 Glu Pro Ala Val Arg Asn Phe Thr Val Thr Glu Lys Gly Trp Glu Phe 755 760 765 Val Trp Gly Asn His Pro Pro Lys Arg Phe Trp Ala Gln Glu Thr Ala 770 775 780 Pro Gly Asn Pro His Gly Leu Pro His Glu Val Ile Thr His Tyr Tyr 785 790 795 800 His Arg Tyr Pro Met Ser Thr Ile Leu Gly Leu Ser Ile Cys Ala Ala 805 810 815 Ile Ala Thr Val Ser Val Ala Ala Ser Thr Trp Leu Phe Cys Arg Ser 820 825 830 Arg Val Ala Cys Leu Thr Pro Tyr Arg Leu Thr Pro Asn Ala Arg Ile 835 840 845 Pro Phe Cys Leu Ala Val Leu Cys Cys Ala Arg Thr Ala Arg Ala Glu 850 855 860 Thr Thr Trp Glu Ser Leu Asp His Leu Trp Asn Asn Asn Gln Gln Met 865 870 875 880 Phe Trp Ile Gln Leu Leu Ile Pro Leu Ala Ala Leu Ile Val Val Thr 885 890 895 Arg Leu Leu Arg Cys Val Cys Cys Val Val Pro Phe Leu Val Met Ala 900 905 910 Gly Ala Ala Gly Ala Gly Ala Tyr Glu His Ala Thr Thr Met Pro Ser 915 920 925 Gln Ala Gly Ile Ser Tyr Asn Thr Ile Val Asn Arg Ala Gly Tyr Ala 930 935 940 Pro Leu Pro Ile Ser Ile Thr Pro Thr Lys Ile Lys Leu Ile Pro Thr 945 950 955 960 Val Asn Leu Glu Tyr Val Thr Cys His Tyr Lys Thr Gly Met Asp Ser 965 970 975 Pro Ala Ile Lys Cys Cys Gly Ser Gln Glu Cys Thr Pro Thr Tyr Arg 980 985 990 Pro Asp Glu Gln Cys Lys Val Phe Thr Gly Val Tyr Pro Phe Met Trp 995 1000 1005 Gly Gly Ala Tyr Cys Phe Cys Asp Thr Glu Asn Thr Gln Val Ser 1010 1015 1020 Lys Ala Tyr Val Met Lys Ser Asp Asp Cys Leu Ala Asp His Ala 1025 1030 1035 Glu Ala Tyr Lys Ala His Thr Ala Ser Val Gln Ala Phe Leu Asn 1040 1045 1050 Ile Thr Val Gly Glu His Ser Ile Val Thr Thr Val Tyr

Val Asn 1055 1060 1065 Gly Glu Thr Pro Val Asn Phe Asn Gly Val Lys Ile Thr Ala Gly 1070 1075 1080 Pro Leu Ser Thr Ala Trp Thr Pro Phe Asp Arg Lys Ile Val Gln 1085 1090 1095 Tyr Ala Gly Glu Ile Tyr Asn Tyr Asp Phe Pro Glu Tyr Gly Ala 1100 1105 1110 Gly Gln Pro Gly Ala Phe Gly Asp Ile Gln Ser Arg Thr Val Ser 1115 1120 1125 Ser Ser Asp Leu Tyr Ala Asn Thr Asn Leu Val Leu Gln Arg Pro 1130 1135 1140 Lys Ala Gly Ala Ile His Val Pro Tyr Thr Gln Ala Pro Ser Gly 1145 1150 1155 Phe Glu Gln Trp Lys Lys Asp Lys Ala Pro Ser Leu Lys Phe Thr 1160 1165 1170 Ala Pro Phe Gly Cys Glu Ile Tyr Thr Asn Pro Ile Arg Ala Glu 1175 1180 1185 Asn Cys Ala Val Gly Ser Ile Pro Leu Ala Phe Asp Ile Pro Asp 1190 1195 1200 Ala Leu Phe Thr Arg Val Ser Glu Thr Pro Thr Leu Ser Ala Ala 1205 1210 1215 Glu Cys Thr Leu Asn Glu Cys Val Tyr Ser Ser Asp Phe Gly Gly 1220 1225 1230 Ile Ala Thr Val Lys Tyr Ser Ala Ser Lys Ser Gly Lys Cys Ala 1235 1240 1245 Val His Val Pro Ser Gly Thr Ala Thr Leu Lys Glu Ala Ala Val 1250 1255 1260 Glu Leu Thr Glu Gln Gly Ser Ala Thr Ile His Phe Ser Thr Ala 1265 1270 1275 Asn Ile His Pro Glu Phe Arg Leu Gln Ile Cys Thr Ser Tyr Val 1280 1285 1290 Thr Cys Lys Gly Asp Cys His Pro Pro Lys Asp His Ile Val Thr 1295 1300 1305 His Pro Gln Tyr His Ala Gln Thr Phe Thr Ala Ala Val Ser Lys 1310 1315 1320 Thr Ala Trp Thr Trp Leu Thr Ser Leu Leu Gly Gly Ser Ala Val 1325 1330 1335 Ile Ile Ile Ile Gly Leu Val Leu Ala Thr Ile Val Ala Met Tyr 1340 1345 1350 Val Leu Thr Asn Gln Lys His Asn 1355 1360 358509DNAArtificial SequenceSynthetic VLP74_21 (VEEV VLP 519 NPNAx6) sequence 35gaattcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt 60ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg 120gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 180ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 240atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 300gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 360gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 420tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 480atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 540gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 600tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga 660gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 720agaagacacc gggaccgatc cagcctccgt taacggtgga gggcagtgta gtctgagcag 780tactcgttgc tgccgcgcgc gccaccagac ataatagctg acagactaac agactgttcc 840tttccatggg tcttttctgc agtcaccgtc gtcgacacgt gtgatcagat atcgcggccg 900ccaccatgtt cccgttccag ccaatgtatc cgatgcagcc aatgccctat cgcaacccgt 960tcgcggcccc gcgcaggccc tggttcccca gaaccgaccc ttttctggcg atgcaggtgc 1020aggaattaac ccgctcgatg gctaacctga cgttcaagca acgccgggac gcgccacctg 1080aggggccatc cgctaataaa ccgaagaagg aggcctcgca aaaacagaaa gggggaggcc 1140aagggaagaa gaagaagaac caagggaaga agaaggctaa gacagggccg cctaatccga 1200aggcacagaa tggaaacaag aagaagacca acaagaaacc aggcaagaga cagcgcatgg 1260tcatgaaatt ggaatctgac aagacgttcc caatcatgtt ggaagggaag ataaacggct 1320acgcttgtgt ggtcggaggg aagttattca ggccgatgca tgtggaaggc aagatcgaca 1380acgacgttct ggccgcgctt aagacgaaga aagcatccaa atacgatctt gagtatgcag 1440atgtgccaca gaacatgcgg gccgatacat tcaaatacac ccatgagaaa ccccaaggct 1500attacagctg gcatcatgga gcagtccaat atgaaaatgg gcgtttcacg gtgccgaaag 1560gagttggggc caagggagac agcggacgac ccattctgga taaccaggga cgggtggtcg 1620ctattgtgct gggaggtgtg aatgaaggat ctaggacagc cctttcagtc gtcatgtgga 1680acgagaaggg agttaccgtg aagtatactc cagagaactg cgagcaatgg tcactagtga 1740ccaccatgtg tctgctcgcc aatgtgacgt tcccatgtgc tcaaccacca atttgctacg 1800acagaaaacc agcagagact ttggccatgc tcagcgttaa cgttgacaac ccgggctacg 1860atgagctgct ggaagcagct gttaagtgcc ccggaaggaa aaggagatcc accgaggagc 1920tgtttaatga gtataagcta acgcgccctt acatggccag atgcatcaga tgtgcagttg 1980ggagctgcca tagtccaata gcaatcgagg cagtaaagag cgacgggcac gacggttatg 2040ttagacttca gacttcctcg cagtatggcc tggattcctc cggcaactta aagggcagga 2100ccatgcggta tgacatgcac gggaccatta aagagatacc actacatcaa gtgtcactct 2160atacatctcg cccgtgtcac attgtggatg ggcacggtta tttcctgctt gccaggtgcc 2220cggcagggga ctccatcacc atggaattta agaaagattc cgtcagacac tcctgctcgg 2280tgccgtatga agtgaaattt aatcctgtag gcagagaact ctatactcat cccccagaac 2340acggagtaga gcaagcgtgc caagtctacg cacatgatgc acagaacaga ggagcttatg 2400tcgagatgca cctcccgggc tcagaagtgg acagcagttt ggtttccttg agcggcagtt 2460ccggaggaaa cccgaatgcc aatcccaacg cgaaccccaa tgctaaccca aatgccaacc 2520caaacgccaa ccccaacgct ggtggatcct cagtcaccgt gacacctcct gatgggacta 2580gcgccctggt ggaatgcgag tgtggcggca caaagatctc cgagaccatc aacaagacaa 2640aacagttcag ccagtgcaca aagaaggagc agtgcagagc atatcggctg cagaacgata 2700agtgggtgta taattctgac aaactgccca aagcagcggg agccacctta aaaggaaaac 2760tgcatgtccc attcttgctg gcagacggca aatgcaccgt gcctctagca ccagaaccta 2820tgataacctt cggtttcaga tcagtgtcac tgaaactgca ccctaagaat cccacatatc 2880taatcacccg ccaacttgct gatgagcctc actacacgca cgagctcata tctgaaccag 2940ctgttaggaa ttttaccgtc accgaaaaag ggtgggagtt tgtatgggga aaccacccgc 3000cgaaaaggtt ttgggcacag gaaacagcac ccggaaatcc acatgggcta ccgcacgagg 3060tgataactca ttattaccac agatacccta tgtccaccat cctgggtttg tcaatttgtg 3120ccgccattgc aaccgtttcc gttgcagcgt ctacctggct gttttgcaga tcaagagttg 3180cgtgcctaac tccttaccgg ctaacaccta acgctaggat accattttgt ctggctgtgc 3240tttgctgcgc ccgcactgcc cgggccgaga ccacctggga gtccttggat cacctatgga 3300acaataacca acagatgttc tggattcaat tgctgatccc tctggccgcc ttgatcgtag 3360tgactcgcct gctcaggtgc gtgtgctgtg tcgtgccttt tttagtcatg gccggcgccg 3420caggcgccgg cgcctacgag cacgcgacca cgatgccgag ccaagcggga atctcgtata 3480acactatagt caacagagca ggctacgcac cactccctat cagcataaca ccaacaaaga 3540tcaagctgat acctacagtg aacttggagt acgtcacctg ccactacaaa acaggaatgg 3600attcaccagc catcaaatgc tgcggatctc aggaatgcac tccaacttac aggcctgatg 3660aacagtgcaa agtcttcaca ggggtttacc cgttcatgtg gggtggtgca tattgctttt 3720gcgacactga gaacacccaa gtcagcaagg cctacgtaat gaaatctgac gactgccttg 3780cggatcatgc tgaagcatat aaagcgcaca cagcctcagt gcaggcgttc ctcaacatca 3840cagtgggaga acactctatt gtgactaccg tgtatgtgaa tggagaaact cctgtgaatt 3900tcaatggggt caaaataact gcaggtccgc tttccacagc ttggacaccc tttgatcgca 3960aaatcgtgca gtatgccggg gagatctata attatgattt tcctgagtat ggggcaggac 4020aaccaggagc atttggagat atacaatcca gaacagtctc aagctctgat ctgtatgcca 4080ataccaacct agtgctgcag agacccaaag caggagcgat ccacgtgcca tacactcagg 4140caccttcggg ttttgagcaa tggaagaaag ataaagctcc atcattgaaa tttaccgccc 4200ctttcggatg cgaaatatat acaaacccca ttcgcgccga aaactgtgct gtagggtcaa 4260ttccattagc ctttgacatt cccgacgcct tgttcaccag ggtgtcagaa acaccgacac 4320tttcagcggc cgaatgcact cttaacgagt gcgtgtattc ttccgacttt ggtgggatcg 4380ccacggtcaa gtactcggcc agcaagtcag gcaagtgcgc agtccatgtg ccatcaggga 4440ctgctaccct aaaagaagca gcagtcgagc taaccgagca agggtcggcg actatccatt 4500tctcgaccgc aaatatccac ccggagttca ggctccaaat atgcacatca tatgttacgt 4560gcaaaggtga ttgtcacccc ccgaaagacc atattgtgac acaccctcag tatcacgccc 4620aaacatttac agccgcggtg tcaaaaaccg cgtggacgtg gttaacatcc ctgctgggag 4680gatcagccgt aattattata attggcttgg tgctggctac tattgtggcc atgtacgtgc 4740tgaccaacca gaaacataat taaggatcta gatctgctgt gccttctagt tgccagccat 4800ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc 4860tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg 4920ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg 4980gggatgcggt gggctctatg ggtacccagg tgctgaagaa ttgacccggt tcctcctggg 5040ccagaaagaa gcaggcacat ccccttctct gtgacacacc ctgtccacgc ccctggttct 5100tagttccagc cccactcata ggacactcat agctcaggag ggctccgcct tcaatcccac 5160ccgctaaagt acttggagcg gtctctccct ccctcatcag cccaccaaac caaacctagc 5220ctccaagagt gggaagaaat taaagcaaga taggctatta agtgcagagg gagagaaaat 5280gcctccaaca tgtgaggaag taatgagaga aatcatagaa ttttaaggcc atgatttaag 5340gccatcatgg cctaagcttg aaaggagata ggatcaaagc ttggcgtaat catggtcata 5400gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 5460cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 5520ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 5580acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 5640gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 5700gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 5760ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 5820cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 5880ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 5940taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 6000ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 6060ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 6120aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 6180tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 6240agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 6300ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 6360tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 6420tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 6480cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 6540aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 6600atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 6660cttaccatct ggccccagtg ctgcaatgat accgcgagaa ccacgctcac cggctccaga 6720tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 6780atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 6840taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 6900tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 6960gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 7020cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 7080cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 7140gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 7200aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 7260accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 7320ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 7380gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 7440aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 7500taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac 7560cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgggtcgc 7620gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccgttga cggtcacagc 7680ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg 7740cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca 7800taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 7860attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga 7920gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 7980caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 8040caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 8100cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 8160agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 8220cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct ttgacgtatg 8280cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca 8340ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag 8400ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag 8460tcacgacgtt gtaaaacgac ggccagtgaa ttccatggtc tcaactttc 8509363858DNAArtificial SequenceSynthetic VLP74_21 (VEEV VLP 519 NPNAx6) VEEV-NPNAx6 sequence 36atgttcccgt tccagccaat gtatccgatg cagccaatgc cctatcgcaa cccgttcgcg 60gccccgcgca ggccctggtt ccccagaacc gacccttttc tggcgatgca ggtgcaggaa 120ttaacccgct cgatggctaa cctgacgttc aagcaacgcc gggacgcgcc acctgagggg 180ccatccgcta ataaaccgaa gaaggaggcc tcgcaaaaac agaaaggggg aggccaaggg 240aagaagaaga agaaccaagg gaagaagaag gctaagacag ggccgcctaa tccgaaggca 300cagaatggaa acaagaagaa gaccaacaag aaaccaggca agagacagcg catggtcatg 360aaattggaat ctgacaagac gttcccaatc atgttggaag ggaagataaa cggctacgct 420tgtgtggtcg gagggaagtt attcaggccg atgcatgtgg aaggcaagat cgacaacgac 480gttctggccg cgcttaagac gaagaaagca tccaaatacg atcttgagta tgcagatgtg 540ccacagaaca tgcgggccga tacattcaaa tacacccatg agaaacccca aggctattac 600agctggcatc atggagcagt ccaatatgaa aatgggcgtt tcacggtgcc gaaaggagtt 660ggggccaagg gagacagcgg acgacccatt ctggataacc agggacgggt ggtcgctatt 720gtgctgggag gtgtgaatga aggatctagg acagcccttt cagtcgtcat gtggaacgag 780aagggagtta ccgtgaagta tactccagag aactgcgagc aatggtcact agtgaccacc 840atgtgtctgc tcgccaatgt gacgttccca tgtgctcaac caccaatttg ctacgacaga 900aaaccagcag agactttggc catgctcagc gttaacgttg acaacccggg ctacgatgag 960ctgctggaag cagctgttaa gtgccccgga aggaaaagga gatccaccga ggagctgttt 1020aatgagtata agctaacgcg cccttacatg gccagatgca tcagatgtgc agttgggagc 1080tgccatagtc caatagcaat cgaggcagta aagagcgacg ggcacgacgg ttatgttaga 1140cttcagactt cctcgcagta tggcctggat tcctccggca acttaaaggg caggaccatg 1200cggtatgaca tgcacgggac cattaaagag ataccactac atcaagtgtc actctataca 1260tctcgcccgt gtcacattgt ggatgggcac ggttatttcc tgcttgccag gtgcccggca 1320ggggactcca tcaccatgga atttaagaaa gattccgtca gacactcctg ctcggtgccg 1380tatgaagtga aatttaatcc tgtaggcaga gaactctata ctcatccccc agaacacgga 1440gtagagcaag cgtgccaagt ctacgcacat gatgcacaga acagaggagc ttatgtcgag 1500atgcacctcc cgggctcaga agtggacagc agtttggttt ccttgagcgg cagttccgga 1560ggaaacccga atgccaatcc caacgcgaac cccaatgcta acccaaatgc caacccaaac 1620gccaacccca acgctggtgg atcctcagtc accgtgacac ctcctgatgg gactagcgcc 1680ctggtggaat gcgagtgtgg cggcacaaag atctccgaga ccatcaacaa gacaaaacag 1740ttcagccagt gcacaaagaa ggagcagtgc agagcatatc ggctgcagaa cgataagtgg 1800gtgtataatt ctgacaaact gcccaaagca gcgggagcca ccttaaaagg aaaactgcat 1860gtcccattct tgctggcaga cggcaaatgc accgtgcctc tagcaccaga acctatgata 1920accttcggtt tcagatcagt gtcactgaaa ctgcacccta agaatcccac atatctaatc 1980acccgccaac ttgctgatga gcctcactac acgcacgagc tcatatctga accagctgtt 2040aggaatttta ccgtcaccga aaaagggtgg gagtttgtat ggggaaacca cccgccgaaa 2100aggttttggg cacaggaaac agcacccgga aatccacatg ggctaccgca cgaggtgata 2160actcattatt accacagata ccctatgtcc accatcctgg gtttgtcaat ttgtgccgcc 2220attgcaaccg tttccgttgc agcgtctacc tggctgtttt gcagatcaag agttgcgtgc 2280ctaactcctt accggctaac acctaacgct aggataccat tttgtctggc tgtgctttgc 2340tgcgcccgca ctgcccgggc cgagaccacc tgggagtcct tggatcacct atggaacaat 2400aaccaacaga tgttctggat tcaattgctg atccctctgg ccgccttgat cgtagtgact 2460cgcctgctca ggtgcgtgtg ctgtgtcgtg ccttttttag tcatggccgg cgccgcaggc 2520gccggcgcct acgagcacgc gaccacgatg ccgagccaag cgggaatctc gtataacact 2580atagtcaaca gagcaggcta cgcaccactc cctatcagca taacaccaac aaagatcaag 2640ctgataccta cagtgaactt ggagtacgtc acctgccact acaaaacagg aatggattca 2700ccagccatca aatgctgcgg atctcaggaa tgcactccaa cttacaggcc tgatgaacag 2760tgcaaagtct tcacaggggt ttacccgttc atgtggggtg gtgcatattg cttttgcgac 2820actgagaaca cccaagtcag caaggcctac gtaatgaaat ctgacgactg ccttgcggat 2880catgctgaag catataaagc gcacacagcc tcagtgcagg cgttcctcaa catcacagtg 2940ggagaacact ctattgtgac taccgtgtat gtgaatggag aaactcctgt gaatttcaat 3000ggggtcaaaa taactgcagg tccgctttcc acagcttgga caccctttga tcgcaaaatc 3060gtgcagtatg ccggggagat ctataattat gattttcctg agtatggggc aggacaacca 3120ggagcatttg gagatataca atccagaaca gtctcaagct ctgatctgta tgccaatacc 3180aacctagtgc tgcagagacc caaagcagga gcgatccacg tgccatacac tcaggcacct 3240tcgggttttg agcaatggaa gaaagataaa gctccatcat tgaaatttac cgcccctttc 3300ggatgcgaaa tatatacaaa ccccattcgc gccgaaaact gtgctgtagg gtcaattcca 3360ttagcctttg acattcccga cgccttgttc accagggtgt cagaaacacc gacactttca 3420gcggccgaat gcactcttaa cgagtgcgtg tattcttccg actttggtgg gatcgccacg 3480gtcaagtact cggccagcaa gtcaggcaag tgcgcagtcc atgtgccatc agggactgct 3540accctaaaag aagcagcagt cgagctaacc gagcaagggt cggcgactat ccatttctcg 3600accgcaaata tccacccgga gttcaggctc caaatatgca catcatatgt tacgtgcaaa 3660ggtgattgtc accccccgaa agaccatatt gtgacacacc ctcagtatca cgcccaaaca 3720tttacagccg cggtgtcaaa aaccgcgtgg acgtggttaa catccctgct gggaggatca 3780gccgtaatta ttataattgg cttggtgctg gctactattg tggccatgta cgtgctgacc 3840aaccagaaac ataattaa 3858371285PRTArtificial SequenceSynthetic VLP74_21 (VEEV VLP 519 NPNAx6) sequence 37Met Phe Pro Phe Gln Pro Met Tyr Pro Met Gln Pro Met Pro Tyr Arg 1 5 10 15 Asn Pro Phe Ala Ala Pro Arg Arg Pro Trp Phe Pro Arg Thr Asp Pro 20 25 30 Phe Leu Ala Met Gln Val Gln Glu Leu Thr Arg Ser Met Ala Asn Leu 35 40 45 Thr Phe Lys Gln

Arg Arg Asp Ala Pro Pro Glu Gly Pro Ser Ala Asn 50 55 60 Lys Pro Lys Lys Glu Ala Ser Gln Lys Gln Lys Gly Gly Gly Gln Gly 65 70 75 80 Lys Lys Lys Lys Asn Gln Gly Lys Lys Lys Ala Lys Thr Gly Pro Pro 85 90 95 Asn Pro Lys Ala Gln Asn Gly Asn Lys Lys Lys Thr Asn Lys Lys Pro 100 105 110 Gly Lys Arg Gln Arg Met Val Met Lys Leu Glu Ser Asp Lys Thr Phe 115 120 125 Pro Ile Met Leu Glu Gly Lys Ile Asn Gly Tyr Ala Cys Val Val Gly 130 135 140 Gly Lys Leu Phe Arg Pro Met His Val Glu Gly Lys Ile Asp Asn Asp 145 150 155 160 Val Leu Ala Ala Leu Lys Thr Lys Lys Ala Ser Lys Tyr Asp Leu Glu 165 170 175 Tyr Ala Asp Val Pro Gln Asn Met Arg Ala Asp Thr Phe Lys Tyr Thr 180 185 190 His Glu Lys Pro Gln Gly Tyr Tyr Ser Trp His His Gly Ala Val Gln 195 200 205 Tyr Glu Asn Gly Arg Phe Thr Val Pro Lys Gly Val Gly Ala Lys Gly 210 215 220 Asp Ser Gly Arg Pro Ile Leu Asp Asn Gln Gly Arg Val Val Ala Ile 225 230 235 240 Val Leu Gly Gly Val Asn Glu Gly Ser Arg Thr Ala Leu Ser Val Val 245 250 255 Met Trp Asn Glu Lys Gly Val Thr Val Lys Tyr Thr Pro Glu Asn Cys 260 265 270 Glu Gln Trp Ser Leu Val Thr Thr Met Cys Leu Leu Ala Asn Val Thr 275 280 285 Phe Pro Cys Ala Gln Pro Pro Ile Cys Tyr Asp Arg Lys Pro Ala Glu 290 295 300 Thr Leu Ala Met Leu Ser Val Asn Val Asp Asn Pro Gly Tyr Asp Glu 305 310 315 320 Leu Leu Glu Ala Ala Val Lys Cys Pro Gly Arg Lys Arg Arg Ser Thr 325 330 335 Glu Glu Leu Phe Asn Glu Tyr Lys Leu Thr Arg Pro Tyr Met Ala Arg 340 345 350 Cys Ile Arg Cys Ala Val Gly Ser Cys His Ser Pro Ile Ala Ile Glu 355 360 365 Ala Val Lys Ser Asp Gly His Asp Gly Tyr Val Arg Leu Gln Thr Ser 370 375 380 Ser Gln Tyr Gly Leu Asp Ser Ser Gly Asn Leu Lys Gly Arg Thr Met 385 390 395 400 Arg Tyr Asp Met His Gly Thr Ile Lys Glu Ile Pro Leu His Gln Val 405 410 415 Ser Leu Tyr Thr Ser Arg Pro Cys His Ile Val Asp Gly His Gly Tyr 420 425 430 Phe Leu Leu Ala Arg Cys Pro Ala Gly Asp Ser Ile Thr Met Glu Phe 435 440 445 Lys Lys Asp Ser Val Arg His Ser Cys Ser Val Pro Tyr Glu Val Lys 450 455 460 Phe Asn Pro Val Gly Arg Glu Leu Tyr Thr His Pro Pro Glu His Gly 465 470 475 480 Val Glu Gln Ala Cys Gln Val Tyr Ala His Asp Ala Gln Asn Arg Gly 485 490 495 Ala Tyr Val Glu Met His Leu Pro Gly Ser Glu Val Asp Ser Ser Leu 500 505 510 Val Ser Leu Ser Gly Ser Ser Gly Gly Asn Pro Asn Ala Asn Pro Asn 515 520 525 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 530 535 540 Ala Gly Gly Ser Ser Val Thr Val Thr Pro Pro Asp Gly Thr Ser Ala 545 550 555 560 Leu Val Glu Cys Glu Cys Gly Gly Thr Lys Ile Ser Glu Thr Ile Asn 565 570 575 Lys Thr Lys Gln Phe Ser Gln Cys Thr Lys Lys Glu Gln Cys Arg Ala 580 585 590 Tyr Arg Leu Gln Asn Asp Lys Trp Val Tyr Asn Ser Asp Lys Leu Pro 595 600 605 Lys Ala Ala Gly Ala Thr Leu Lys Gly Lys Leu His Val Pro Phe Leu 610 615 620 Leu Ala Asp Gly Lys Cys Thr Val Pro Leu Ala Pro Glu Pro Met Ile 625 630 635 640 Thr Phe Gly Phe Arg Ser Val Ser Leu Lys Leu His Pro Lys Asn Pro 645 650 655 Thr Tyr Leu Ile Thr Arg Gln Leu Ala Asp Glu Pro His Tyr Thr His 660 665 670 Glu Leu Ile Ser Glu Pro Ala Val Arg Asn Phe Thr Val Thr Glu Lys 675 680 685 Gly Trp Glu Phe Val Trp Gly Asn His Pro Pro Lys Arg Phe Trp Ala 690 695 700 Gln Glu Thr Ala Pro Gly Asn Pro His Gly Leu Pro His Glu Val Ile 705 710 715 720 Thr His Tyr Tyr His Arg Tyr Pro Met Ser Thr Ile Leu Gly Leu Ser 725 730 735 Ile Cys Ala Ala Ile Ala Thr Val Ser Val Ala Ala Ser Thr Trp Leu 740 745 750 Phe Cys Arg Ser Arg Val Ala Cys Leu Thr Pro Tyr Arg Leu Thr Pro 755 760 765 Asn Ala Arg Ile Pro Phe Cys Leu Ala Val Leu Cys Cys Ala Arg Thr 770 775 780 Ala Arg Ala Glu Thr Thr Trp Glu Ser Leu Asp His Leu Trp Asn Asn 785 790 795 800 Asn Gln Gln Met Phe Trp Ile Gln Leu Leu Ile Pro Leu Ala Ala Leu 805 810 815 Ile Val Val Thr Arg Leu Leu Arg Cys Val Cys Cys Val Val Pro Phe 820 825 830 Leu Val Met Ala Gly Ala Ala Gly Ala Gly Ala Tyr Glu His Ala Thr 835 840 845 Thr Met Pro Ser Gln Ala Gly Ile Ser Tyr Asn Thr Ile Val Asn Arg 850 855 860 Ala Gly Tyr Ala Pro Leu Pro Ile Ser Ile Thr Pro Thr Lys Ile Lys 865 870 875 880 Leu Ile Pro Thr Val Asn Leu Glu Tyr Val Thr Cys His Tyr Lys Thr 885 890 895 Gly Met Asp Ser Pro Ala Ile Lys Cys Cys Gly Ser Gln Glu Cys Thr 900 905 910 Pro Thr Tyr Arg Pro Asp Glu Gln Cys Lys Val Phe Thr Gly Val Tyr 915 920 925 Pro Phe Met Trp Gly Gly Ala Tyr Cys Phe Cys Asp Thr Glu Asn Thr 930 935 940 Gln Val Ser Lys Ala Tyr Val Met Lys Ser Asp Asp Cys Leu Ala Asp 945 950 955 960 His Ala Glu Ala Tyr Lys Ala His Thr Ala Ser Val Gln Ala Phe Leu 965 970 975 Asn Ile Thr Val Gly Glu His Ser Ile Val Thr Thr Val Tyr Val Asn 980 985 990 Gly Glu Thr Pro Val Asn Phe Asn Gly Val Lys Ile Thr Ala Gly Pro 995 1000 1005 Leu Ser Thr Ala Trp Thr Pro Phe Asp Arg Lys Ile Val Gln Tyr 1010 1015 1020 Ala Gly Glu Ile Tyr Asn Tyr Asp Phe Pro Glu Tyr Gly Ala Gly 1025 1030 1035 Gln Pro Gly Ala Phe Gly Asp Ile Gln Ser Arg Thr Val Ser Ser 1040 1045 1050 Ser Asp Leu Tyr Ala Asn Thr Asn Leu Val Leu Gln Arg Pro Lys 1055 1060 1065 Ala Gly Ala Ile His Val Pro Tyr Thr Gln Ala Pro Ser Gly Phe 1070 1075 1080 Glu Gln Trp Lys Lys Asp Lys Ala Pro Ser Leu Lys Phe Thr Ala 1085 1090 1095 Pro Phe Gly Cys Glu Ile Tyr Thr Asn Pro Ile Arg Ala Glu Asn 1100 1105 1110 Cys Ala Val Gly Ser Ile Pro Leu Ala Phe Asp Ile Pro Asp Ala 1115 1120 1125 Leu Phe Thr Arg Val Ser Glu Thr Pro Thr Leu Ser Ala Ala Glu 1130 1135 1140 Cys Thr Leu Asn Glu Cys Val Tyr Ser Ser Asp Phe Gly Gly Ile 1145 1150 1155 Ala Thr Val Lys Tyr Ser Ala Ser Lys Ser Gly Lys Cys Ala Val 1160 1165 1170 His Val Pro Ser Gly Thr Ala Thr Leu Lys Glu Ala Ala Val Glu 1175 1180 1185 Leu Thr Glu Gln Gly Ser Ala Thr Ile His Phe Ser Thr Ala Asn 1190 1195 1200 Ile His Pro Glu Phe Arg Leu Gln Ile Cys Thr Ser Tyr Val Thr 1205 1210 1215 Cys Lys Gly Asp Cys His Pro Pro Lys Asp His Ile Val Thr His 1220 1225 1230 Pro Gln Tyr His Ala Gln Thr Phe Thr Ala Ala Val Ser Lys Thr 1235 1240 1245 Ala Trp Thr Trp Leu Thr Ser Leu Leu Gly Gly Ser Ala Val Ile 1250 1255 1260 Ile Ile Ile Gly Leu Val Leu Ala Thr Ile Val Ala Met Tyr Val 1265 1270 1275 Leu Thr Asn Gln Lys His Asn 1280 1285 387305DNAArtificial SequenceSynthetic VLP74_15 (CHI VLP 532 NPNAx6) sequence 38tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 240attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 300tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 360tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtgtt ggcattgatt 420attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga 480gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg 540cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg 600acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca 660tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 720ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc 780tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc 840acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 900tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag 960gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agataactgc 1020aggtcgacga tatcgcggcc gccaccatgg agttcatccc gacgcaaact ttctataaca 1080gaaggtacca accccgaccc tgggccccac gccctacaat tcaagtaatt agacctagac 1140cacgtccaca gaggcaggct gggcaactcg cccagctgat ctccgcagtc aacaaattga 1200ccatgcgcgc ggtacctcaa cagaagcctc gcagaaatcg gaaaaacaag aagcaaaggc 1260agaagaagca ggcgccgcaa aacgacccaa agcaaaagaa gcaaccacca caaaagaagc 1320cggctcaaaa gaagaagaaa ccaggccgta gggagagaat gtgcatgaaa attgaaaatg 1380attgcatctt cgaagtcaag catgaaggca aagtgatggg ctacgcatgc ctggtggggg 1440ataaagtaat gaaaccagca catgtgaagg gaactatcga caatgccgat ctggctaaac 1500tggcctttaa gcggtcgtct aaatacgatc ttgaatgtgc acagataccg gtgcacatga 1560agtctgatgc ctcgaagttt acccacgaga aacccgaggg gtactataac tggcatcacg 1620gagcagtgca gtattcagga ggccggttca ctatcccgac gggtgcaggc aagccgggag 1680acagcggcag accgatcttc gacaacaaag gacgggtggt ggccatcgtc ctaggagggg 1740ccaacgaagg tgcccgcacg gccctctccg tggtgacgtg gaacaaagac atcgtcacaa 1800aaattacccc tgagggagcc gaagagtgga gcctcgccct cccggtcttg tgcctgttgg 1860caaacactac attcccctgc tctcagccgc cttgcacacc ctgctgctac gaaaaggaac 1920cggaaagcac cttgcgcatg cttgaggaca acgtgatgag acccggatac taccagctac 1980taaaagcatc gctgacttgc tctccccacc gccaaagacg cagtactaag gacaatttta 2040atgtctataa agccacaaga ccatatctag ctcattgtcc tgactgcgga gaagggcatt 2100cgtgccacag ccctatcgca ttggagcgca tcagaaatga agcaacggac ggaacgctga 2160aaatccaggt ctctttgcag atcgggataa agacagatga cagccacgat tggaccaagc 2220tgcgctatat ggatagccat acgccagcgg acgcggagcg agccggattg cttgtaagga 2280cttcagcacc gtgcacgatc accgggacca tgggacactt tattctcgcc cgatgcccga 2340aaggagagac gctgacagtg ggatttacgg acagcagaaa gatcagccac acatgcacac 2400acccgttcca tcatgaacca cctgtgatag gtagggagag gttccactct cgaccacaac 2460atggtaaaga gttaccttgc agcacgtacg tgcagagcac cgctgccact gctgaggaga 2520tagaggtgca tatgccccca gatactcctg accgcacgct gatgacgcag cagtctggca 2580acgtgaagat cacagttaat gggcagacgg tgcggtacaa gtgcaactgc ggtggctccg 2640gaggaaaccc gaatgccaat cccaacgcga accccaatgc taacccaaat gccaacccaa 2700acgccaaccc caacgctggt ggatccaacg agggactgac aaccacagac aaagtgatca 2760ataactgcaa aattgatcag tgccatgctg cagtcactaa tcacaagaat tggcaataca 2820actccccttt agtcccgcgc aacgctgaac tcggggaccg taaaggaaag atccacatcc 2880cattcccatt ggcaaacgtg acttgcagag tgccaaaagc aagaaaccct acagtaactt 2940acggaaaaaa ccaagtcacc atgctgctgt atcctgacca tccgacactc ttgtcttacc 3000gtaacatggg acaggaacca aattaccacg aggagtgggt gacacacaag aaggaggtta 3060ccttgaccgt gcctactgag ggtctggagg tcacttgggg caacaacgaa ccatacaagt 3120actggccgca gatgtctacg aacggtactg ctcatggtca cccacatgag ataatcttgt 3180actattatga gctgtacccc actatgactg tagtcattgt gtcggtggcc tcgttcgtgc 3240ttctgtcgat ggtgggcaca gcagtgggaa tgtgtgtgtg cgcacggcgc agatgcatta 3300caccatatga attaacacca ggagccactg ttcccttcct gctcagcctg ctatgctgcg 3360tcagaacgac caaggcggcc acatattacg aggctgcggc atatctatgg aacgaacagc 3420agcccctgtt ctggttgcag gctcttatcc cgctggccgc cttgatcgtc ctgtgcaact 3480gtctgaaact cttgccatgc tgctgtaaga ccctggcttt tttagccgta atgagcatcg 3540gtgcccacac tgtgagcgcg tacgaacacg taacagtgat cccgaacacg gtgggagtac 3600cgtataagac tcttgtcaac agaccgggtt acagccccat ggtgttggag atggagctac 3660aatcagtcac cttggaacca acactgtcac ttgactacat cacgtgcgag tacaaaactg 3720tcatcccctc cccgtacgtg aagtgctgtg gtacagcaga gtgcaaggac aagagcctac 3780cagactacag ctgcaaggtc tttactggag tctacccatt tatgtggggc ggcgcctact 3840gcttttgcga cgccgaaaat acgcaattga gcgaggcaca tgtagagaaa tctgaatctt 3900gcaaaacaga gtttgcatcg gcctacagag cccacaccgc atcggcgtcg gcgaagctcc 3960gcgtccttta ccaaggaaac aacattaccg tagctgccta cgctaacggt gaccatgccg 4020tcacagtaaa ggacgccaag tttgtcgtgg gcccaatgtc ctccgcctgg acaccttttg 4080acaacaaaat cgtggtgtac aaaggcgacg tctacaacat ggactaccca ccttttggcg 4140caggaagacc aggacaattt ggtgacattc aaagtcgtac accggaaagt aaagacgttt 4200atgccaacac tcagttggta ctacagaggc cagcagcagg cacggtacat gtaccatact 4260ctcaggcacc atctggcttc aagtattggc tgaaggaacg aggagcatcg ctacagcaca 4320cggcaccgtt cggttgccag attgcgacaa acccggtaag agctgtaaat tgcgctgtgg 4380ggaacatacc aatttccatc gacataccgg atgcggcctt tactagggtt gtcgatgcac 4440cctctgtaac ggacatgtca tgcgaagtac cagcctgcac tcactcctcc gactttgggg 4500gcgtcgccat catcaaatac acagctagca agaaaggtaa atgtgcagta cattcgatga 4560ccaacgccgt taccattcga gaagccgacg tagaagtaga ggggaactcc cagctgcaaa 4620tatccttctc aacagccctg gcaagcgccg agtttcgcgt gcaagtgtgc tccacacaag 4680tacactgcgc agccgcatgc caccctccaa aggaccacat agtcaattac ccagcatcac 4740acaccaccct tggggtccag gatatatcca caacggcaat gtcttgggtg cagaagatta 4800cgggaggagt aggattaatt gttgctgttg ctgccttaat tttaattgtg gtgctatgcg 4860tgtcgtttag caggcactaa ggatctagat ctgctgtgcc ttctagttgc cagccatctg 4920ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt 4980cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg 5040gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg 5100atgcggtggg ctctatggct cgagcatggt catagctgtt tcctgtgtga aattgttatc 5160cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 5220aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 5280acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 5340ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 5400gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 5460caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 5520tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 5580gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 5640ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 5700cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 5760tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 5820tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 5880cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 5940agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga 6000agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 6060gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 6120aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 6180ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 6240gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt tagaaaaact 6300catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt 6360gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa 6420gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc 6480cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg 6540agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct 6600cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga 6660gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc 6720gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata 6780cctggaatgc tgttttccca gggatcgcag tggtgagtaa

ccatgcatca tcaggagtac 6840ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca 6900tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg 6960catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag 7020cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag 7080acgtttcccg ttgaatatgg ctcatactct tcctttttca atattattga agcatttatc 7140agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 7200gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca 7260tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc 7305391277PRTArtificial SequenceSynthetic VLP74_15 (CHI VLP 532 NPNAx6) sequence 39Met Glu Phe Ile Pro Thr Gln Thr Phe Tyr Asn Arg Arg Tyr Gln Pro 1 5 10 15 Arg Pro Trp Ala Pro Arg Pro Thr Ile Gln Val Ile Arg Pro Arg Pro 20 25 30 Arg Pro Gln Arg Gln Ala Gly Gln Leu Ala Gln Leu Ile Ser Ala Val 35 40 45 Asn Lys Leu Thr Met Arg Ala Val Pro Gln Gln Lys Pro Arg Arg Asn 50 55 60 Arg Lys Asn Lys Lys Gln Arg Gln Lys Lys Gln Ala Pro Gln Asn Asp 65 70 75 80 Pro Lys Gln Lys Lys Gln Pro Pro Gln Lys Lys Pro Ala Gln Lys Lys 85 90 95 Lys Lys Pro Gly Arg Arg Glu Arg Met Cys Met Lys Ile Glu Asn Asp 100 105 110 Cys Ile Phe Glu Val Lys His Glu Gly Lys Val Met Gly Tyr Ala Cys 115 120 125 Leu Val Gly Asp Lys Val Met Lys Pro Ala His Val Lys Gly Thr Ile 130 135 140 Asp Asn Ala Asp Leu Ala Lys Leu Ala Phe Lys Arg Ser Ser Lys Tyr 145 150 155 160 Asp Leu Glu Cys Ala Gln Ile Pro Val His Met Lys Ser Asp Ala Ser 165 170 175 Lys Phe Thr His Glu Lys Pro Glu Gly Tyr Tyr Asn Trp His His Gly 180 185 190 Ala Val Gln Tyr Ser Gly Gly Arg Phe Thr Ile Pro Thr Gly Ala Gly 195 200 205 Lys Pro Gly Asp Ser Gly Arg Pro Ile Phe Asp Asn Lys Gly Arg Val 210 215 220 Val Ala Ile Val Leu Gly Gly Ala Asn Glu Gly Ala Arg Thr Ala Leu 225 230 235 240 Ser Val Val Thr Trp Asn Lys Asp Ile Val Thr Lys Ile Thr Pro Glu 245 250 255 Gly Ala Glu Glu Trp Ser Leu Ala Leu Pro Val Leu Cys Leu Leu Ala 260 265 270 Asn Thr Thr Phe Pro Cys Ser Gln Pro Pro Cys Thr Pro Cys Cys Tyr 275 280 285 Glu Lys Glu Pro Glu Ser Thr Leu Arg Met Leu Glu Asp Asn Val Met 290 295 300 Arg Pro Gly Tyr Tyr Gln Leu Leu Lys Ala Ser Leu Thr Cys Ser Pro 305 310 315 320 His Arg Gln Arg Arg Ser Thr Lys Asp Asn Phe Asn Val Tyr Lys Ala 325 330 335 Thr Arg Pro Tyr Leu Ala His Cys Pro Asp Cys Gly Glu Gly His Ser 340 345 350 Cys His Ser Pro Ile Ala Leu Glu Arg Ile Arg Asn Glu Ala Thr Asp 355 360 365 Gly Thr Leu Lys Ile Gln Val Ser Leu Gln Ile Gly Ile Lys Thr Asp 370 375 380 Asp Ser His Asp Trp Thr Lys Leu Arg Tyr Met Asp Ser His Thr Pro 385 390 395 400 Ala Asp Ala Glu Arg Ala Gly Leu Leu Val Arg Thr Ser Ala Pro Cys 405 410 415 Thr Ile Thr Gly Thr Met Gly His Phe Ile Leu Ala Arg Cys Pro Lys 420 425 430 Gly Glu Thr Leu Thr Val Gly Phe Thr Asp Ser Arg Lys Ile Ser His 435 440 445 Thr Cys Thr His Pro Phe His His Glu Pro Pro Val Ile Gly Arg Glu 450 455 460 Arg Phe His Ser Arg Pro Gln His Gly Lys Glu Leu Pro Cys Ser Thr 465 470 475 480 Tyr Val Gln Ser Thr Ala Ala Thr Ala Glu Glu Ile Glu Val His Met 485 490 495 Pro Pro Asp Thr Pro Asp Arg Thr Leu Met Thr Gln Gln Ser Gly Asn 500 505 510 Val Lys Ile Thr Val Asn Gly Gln Thr Val Arg Tyr Lys Cys Asn Cys 515 520 525 Gly Gly Ser Gly Gly Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 530 535 540 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Gly Gly Ser 545 550 555 560 Asn Glu Gly Leu Thr Thr Thr Asp Lys Val Ile Asn Asn Cys Lys Ile 565 570 575 Asp Gln Cys His Ala Ala Val Thr Asn His Lys Asn Trp Gln Tyr Asn 580 585 590 Ser Pro Leu Val Pro Arg Asn Ala Glu Leu Gly Asp Arg Lys Gly Lys 595 600 605 Ile His Ile Pro Phe Pro Leu Ala Asn Val Thr Cys Arg Val Pro Lys 610 615 620 Ala Arg Asn Pro Thr Val Thr Tyr Gly Lys Asn Gln Val Thr Met Leu 625 630 635 640 Leu Tyr Pro Asp His Pro Thr Leu Leu Ser Tyr Arg Asn Met Gly Gln 645 650 655 Glu Pro Asn Tyr His Glu Glu Trp Val Thr His Lys Lys Glu Val Thr 660 665 670 Leu Thr Val Pro Thr Glu Gly Leu Glu Val Thr Trp Gly Asn Asn Glu 675 680 685 Pro Tyr Lys Tyr Trp Pro Gln Met Ser Thr Asn Gly Thr Ala His Gly 690 695 700 His Pro His Glu Ile Ile Leu Tyr Tyr Tyr Glu Leu Tyr Pro Thr Met 705 710 715 720 Thr Val Val Ile Val Ser Val Ala Ser Phe Val Leu Leu Ser Met Val 725 730 735 Gly Thr Ala Val Gly Met Cys Val Cys Ala Arg Arg Arg Cys Ile Thr 740 745 750 Pro Tyr Glu Leu Thr Pro Gly Ala Thr Val Pro Phe Leu Leu Ser Leu 755 760 765 Leu Cys Cys Val Arg Thr Thr Lys Ala Ala Thr Tyr Tyr Glu Ala Ala 770 775 780 Ala Tyr Leu Trp Asn Glu Gln Gln Pro Leu Phe Trp Leu Gln Ala Leu 785 790 795 800 Ile Pro Leu Ala Ala Leu Ile Val Leu Cys Asn Cys Leu Lys Leu Leu 805 810 815 Pro Cys Cys Cys Lys Thr Leu Ala Phe Leu Ala Val Met Ser Ile Gly 820 825 830 Ala His Thr Val Ser Ala Tyr Glu His Val Thr Val Ile Pro Asn Thr 835 840 845 Val Gly Val Pro Tyr Lys Thr Leu Val Asn Arg Pro Gly Tyr Ser Pro 850 855 860 Met Val Leu Glu Met Glu Leu Gln Ser Val Thr Leu Glu Pro Thr Leu 865 870 875 880 Ser Leu Asp Tyr Ile Thr Cys Glu Tyr Lys Thr Val Ile Pro Ser Pro 885 890 895 Tyr Val Lys Cys Cys Gly Thr Ala Glu Cys Lys Asp Lys Ser Leu Pro 900 905 910 Asp Tyr Ser Cys Lys Val Phe Thr Gly Val Tyr Pro Phe Met Trp Gly 915 920 925 Gly Ala Tyr Cys Phe Cys Asp Ala Glu Asn Thr Gln Leu Ser Glu Ala 930 935 940 His Val Glu Lys Ser Glu Ser Cys Lys Thr Glu Phe Ala Ser Ala Tyr 945 950 955 960 Arg Ala His Thr Ala Ser Ala Ser Ala Lys Leu Arg Val Leu Tyr Gln 965 970 975 Gly Asn Asn Ile Thr Val Ala Ala Tyr Ala Asn Gly Asp His Ala Val 980 985 990 Thr Val Lys Asp Ala Lys Phe Val Val Gly Pro Met Ser Ser Ala Trp 995 1000 1005 Thr Pro Phe Asp Asn Lys Ile Val Val Tyr Lys Gly Asp Val Tyr 1010 1015 1020 Asn Met Asp Tyr Pro Pro Phe Gly Ala Gly Arg Pro Gly Gln Phe 1025 1030 1035 Gly Asp Ile Gln Ser Arg Thr Pro Glu Ser Lys Asp Val Tyr Ala 1040 1045 1050 Asn Thr Gln Leu Val Leu Gln Arg Pro Ala Ala Gly Thr Val His 1055 1060 1065 Val Pro Tyr Ser Gln Ala Pro Ser Gly Phe Lys Tyr Trp Leu Lys 1070 1075 1080 Glu Arg Gly Ala Ser Leu Gln His Thr Ala Pro Phe Gly Cys Gln 1085 1090 1095 Ile Ala Thr Asn Pro Val Arg Ala Val Asn Cys Ala Val Gly Asn 1100 1105 1110 Ile Pro Ile Ser Ile Asp Ile Pro Asp Ala Ala Phe Thr Arg Val 1115 1120 1125 Val Asp Ala Pro Ser Val Thr Asp Met Ser Cys Glu Val Pro Ala 1130 1135 1140 Cys Thr His Ser Ser Asp Phe Gly Gly Val Ala Ile Ile Lys Tyr 1145 1150 1155 Thr Ala Ser Lys Lys Gly Lys Cys Ala Val His Ser Met Thr Asn 1160 1165 1170 Ala Val Thr Ile Arg Glu Ala Asp Val Glu Val Glu Gly Asn Ser 1175 1180 1185 Gln Leu Gln Ile Ser Phe Ser Thr Ala Leu Ala Ser Ala Glu Phe 1190 1195 1200 Arg Val Gln Val Cys Ser Thr Gln Val His Cys Ala Ala Ala Cys 1205 1210 1215 His Pro Pro Lys Asp His Ile Val Asn Tyr Pro Ala Ser His Thr 1220 1225 1230 Thr Leu Gly Val Gln Asp Ile Ser Thr Thr Ala Met Ser Trp Val 1235 1240 1245 Gln Lys Ile Thr Gly Gly Val Gly Leu Ile Val Ala Val Ala Ala 1250 1255 1260 Leu Ile Leu Ile Val Val Leu Cys Val Ser Phe Ser Arg His 1265 1270 1275 407533DNAArtificial SequenceSynthetic VLP78_15 (CHI VLP 532 NPNAx25) sequence 40tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 240attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 300tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 360tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtgtt ggcattgatt 420attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga 480gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg 540cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg 600acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca 660tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 720ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc 780tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc 840acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 900tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag 960gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agataactgc 1020aggtcgacga tatcgcggcc gccaccatgg agttcatccc gacgcaaact ttctataaca 1080gaaggtacca accccgaccc tgggccccac gccctacaat tcaagtaatt agacctagac 1140cacgtccaca gaggcaggct gggcaactcg cccagctgat ctccgcagtc aacaaattga 1200ccatgcgcgc ggtacctcaa cagaagcctc gcagaaatcg gaaaaacaag aagcaaaggc 1260agaagaagca ggcgccgcaa aacgacccaa agcaaaagaa gcaaccacca caaaagaagc 1320cggctcaaaa gaagaagaaa ccaggccgta gggagagaat gtgcatgaaa attgaaaatg 1380attgcatctt cgaagtcaag catgaaggca aagtgatggg ctacgcatgc ctggtggggg 1440ataaagtaat gaaaccagca catgtgaagg gaactatcga caatgccgat ctggctaaac 1500tggcctttaa gcggtcgtct aaatacgatc ttgaatgtgc acagataccg gtgcacatga 1560agtctgatgc ctcgaagttt acccacgaga aacccgaggg gtactataac tggcatcacg 1620gagcagtgca gtattcagga ggccggttca ctatcccgac gggtgcaggc aagccgggag 1680acagcggcag accgatcttc gacaacaaag gacgggtggt ggccatcgtc ctaggagggg 1740ccaacgaagg tgcccgcacg gccctctccg tggtgacgtg gaacaaagac atcgtcacaa 1800aaattacccc tgagggagcc gaagagtgga gcctcgccct cccggtcttg tgcctgttgg 1860caaacactac attcccctgc tctcagccgc cttgcacacc ctgctgctac gaaaaggaac 1920cggaaagcac cttgcgcatg cttgaggaca acgtgatgag acccggatac taccagctac 1980taaaagcatc gctgacttgc tctccccacc gccaaagacg cagtactaag gacaatttta 2040atgtctataa agccacaaga ccatatctag ctcattgtcc tgactgcgga gaagggcatt 2100cgtgccacag ccctatcgca ttggagcgca tcagaaatga agcaacggac ggaacgctga 2160aaatccaggt ctctttgcag atcgggataa agacagatga cagccacgat tggaccaagc 2220tgcgctatat ggatagccat acgccagcgg acgcggagcg agccggattg cttgtaagga 2280cttcagcacc gtgcacgatc accgggacca tgggacactt tattctcgcc cgatgcccga 2340aaggagagac gctgacagtg ggatttacgg acagcagaaa gatcagccac acatgcacac 2400acccgttcca tcatgaacca cctgtgatag gtagggagag gttccactct cgaccacaac 2460atggtaaaga gttaccttgc agcacgtacg tgcagagcac cgctgccact gctgaggaga 2520tagaggtgca tatgccccca gatactcctg accgcacgct gatgacgcag cagtctggca 2580acgtgaagat cacagttaat gggcagacgg tgcggtacaa gtgcaactgc ggtggctccg 2640gaggaaaccc gaatgccaat cccaacgcga accccaacgc taaccccaac gccaatccga 2700atgcaaaccc gaacgttgac ccaaacgcca acccgaatgc caatcccaac gcgaacccca 2760atgctaaccc aaatgccaac ccaaacgcca accccaacgc taatccaaac gccaacccta 2820acgccaatcc caacgcgaat cctaacgcta atcccaacgc aaatcccaat gctaatccga 2880acgcgaaccc taatgcaaac cccaacgcca acccgaacgc taacccgaac gctaatccca 2940acgccggtgg atccaacgag ggactgacaa ccacagacaa agtgatcaat aactgcaaaa 3000ttgatcagtg ccatgctgca gtcactaatc acaagaattg gcaatacaac tcccctttag 3060tcccgcgcaa cgctgaactc ggggaccgta aaggaaagat ccacatccca ttcccattgg 3120caaacgtgac ttgcagagtg ccaaaagcaa gaaaccctac agtaacttac ggaaaaaacc 3180aagtcaccat gctgctgtat cctgaccatc cgacactctt gtcttaccgt aacatgggac 3240aggaaccaaa ttaccacgag gagtgggtga cacacaagaa ggaggttacc ttgaccgtgc 3300ctactgaggg tctggaggtc acttggggca acaacgaacc atacaagtac tggccgcaga 3360tgtctacgaa cggtactgct catggtcacc cacatgagat aatcttgtac tattatgagc 3420tgtaccccac tatgactgta gtcattgtgt cggtggcctc gttcgtgctt ctgtcgatgg 3480tgggcacagc agtgggaatg tgtgtgtgcg cacggcgcag atgcattaca ccatatgaat 3540taacaccagg agccactgtt cccttcctgc tcagcctgct atgctgcgtc agaacgacca 3600aggcggccac atattacgag gctgcggcat atctatggaa cgaacagcag cccctgttct 3660ggttgcaggc tcttatcccg ctggccgcct tgatcgtcct gtgcaactgt ctgaaactct 3720tgccatgctg ctgtaagacc ctggcttttt tagccgtaat gagcatcggt gcccacactg 3780tgagcgcgta cgaacacgta acagtgatcc cgaacacggt gggagtaccg tataagactc 3840ttgtcaacag accgggttac agccccatgg tgttggagat ggagctacaa tcagtcacct 3900tggaaccaac actgtcactt gactacatca cgtgcgagta caaaactgtc atcccctccc 3960cgtacgtgaa gtgctgtggt acagcagagt gcaaggacaa gagcctacca gactacagct 4020gcaaggtctt tactggagtc tacccattta tgtggggcgg cgcctactgc ttttgcgacg 4080ccgaaaatac gcaattgagc gaggcacatg tagagaaatc tgaatcttgc aaaacagagt 4140ttgcatcggc ctacagagcc cacaccgcat cggcgtcggc gaagctccgc gtcctttacc 4200aaggaaacaa cattaccgta gctgcctacg ctaacggtga ccatgccgtc acagtaaagg 4260acgccaagtt tgtcgtgggc ccaatgtcct ccgcctggac accttttgac aacaaaatcg 4320tggtgtacaa aggcgacgtc tacaacatgg actacccacc ttttggcgca ggaagaccag 4380gacaatttgg tgacattcaa agtcgtacac cggaaagtaa agacgtttat gccaacactc 4440agttggtact acagaggcca gcagcaggca cggtacatgt accatactct caggcaccat 4500ctggcttcaa gtattggctg aaggaacgag gagcatcgct acagcacacg gcaccgttcg 4560gttgccagat tgcgacaaac ccggtaagag ctgtaaattg cgctgtgggg aacataccaa 4620tttccatcga cataccggat gcggccttta ctagggttgt cgatgcaccc tctgtaacgg 4680acatgtcatg cgaagtacca gcctgcactc actcctccga ctttgggggc gtcgccatca 4740tcaaatacac agctagcaag aaaggtaaat gtgcagtaca ttcgatgacc aacgccgtta 4800ccattcgaga agccgacgta gaagtagagg ggaactccca gctgcaaata tccttctcaa 4860cagccctggc aagcgccgag tttcgcgtgc aagtgtgctc cacacaagta cactgcgcag 4920ccgcatgcca ccctccaaag gaccacatag tcaattaccc agcatcacac accacccttg 4980gggtccagga tatatccaca acggcaatgt cttgggtgca gaagattacg ggaggagtag 5040gattaattgt tgctgttgct gccttaattt taattgtggt gctatgcgtg tcgtttagca 5100ggcactaagg atctagatct gctgtgcctt ctagttgcca gccatctgtt gtttgcccct 5160cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg 5220aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc 5280aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct 5340ctatggctcg agcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 5400cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 5460aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 5520agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 5580ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 5640ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 5700tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 5760tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 5820gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 5880ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 5940tggcgctttc tcatagctca cgctgtaggt atctcagttc

ggtgtaggtc gttcgctcca 6000agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 6060atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 6120acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 6180actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 6240tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 6300tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 6360tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 6420tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 6480caatctaaag tatatatgag taaacttggt ctgacagtta gaaaaactca tcgagcatca 6540aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt 6600tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc 6660ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa 6720taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa 6780gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat 6840cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc 6900gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg 6960ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg 7020ttttcccagg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct 7080tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa 7140catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc 7200catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc 7260catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt 7320gaatatggct catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 7380tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 7440catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct 7500ataaaaatag gcgtatcacg aggccctttc gtc 7533411353PRTArtificial SequenceSynthetic VLP78_15 (CHI VLP 532 NPNAx25) sequence 41Met Glu Phe Ile Pro Thr Gln Thr Phe Tyr Asn Arg Arg Tyr Gln Pro 1 5 10 15 Arg Pro Trp Ala Pro Arg Pro Thr Ile Gln Val Ile Arg Pro Arg Pro 20 25 30 Arg Pro Gln Arg Gln Ala Gly Gln Leu Ala Gln Leu Ile Ser Ala Val 35 40 45 Asn Lys Leu Thr Met Arg Ala Val Pro Gln Gln Lys Pro Arg Arg Asn 50 55 60 Arg Lys Asn Lys Lys Gln Arg Gln Lys Lys Gln Ala Pro Gln Asn Asp 65 70 75 80 Pro Lys Gln Lys Lys Gln Pro Pro Gln Lys Lys Pro Ala Gln Lys Lys 85 90 95 Lys Lys Pro Gly Arg Arg Glu Arg Met Cys Met Lys Ile Glu Asn Asp 100 105 110 Cys Ile Phe Glu Val Lys His Glu Gly Lys Val Met Gly Tyr Ala Cys 115 120 125 Leu Val Gly Asp Lys Val Met Lys Pro Ala His Val Lys Gly Thr Ile 130 135 140 Asp Asn Ala Asp Leu Ala Lys Leu Ala Phe Lys Arg Ser Ser Lys Tyr 145 150 155 160 Asp Leu Glu Cys Ala Gln Ile Pro Val His Met Lys Ser Asp Ala Ser 165 170 175 Lys Phe Thr His Glu Lys Pro Glu Gly Tyr Tyr Asn Trp His His Gly 180 185 190 Ala Val Gln Tyr Ser Gly Gly Arg Phe Thr Ile Pro Thr Gly Ala Gly 195 200 205 Lys Pro Gly Asp Ser Gly Arg Pro Ile Phe Asp Asn Lys Gly Arg Val 210 215 220 Val Ala Ile Val Leu Gly Gly Ala Asn Glu Gly Ala Arg Thr Ala Leu 225 230 235 240 Ser Val Val Thr Trp Asn Lys Asp Ile Val Thr Lys Ile Thr Pro Glu 245 250 255 Gly Ala Glu Glu Trp Ser Leu Ala Leu Pro Val Leu Cys Leu Leu Ala 260 265 270 Asn Thr Thr Phe Pro Cys Ser Gln Pro Pro Cys Thr Pro Cys Cys Tyr 275 280 285 Glu Lys Glu Pro Glu Ser Thr Leu Arg Met Leu Glu Asp Asn Val Met 290 295 300 Arg Pro Gly Tyr Tyr Gln Leu Leu Lys Ala Ser Leu Thr Cys Ser Pro 305 310 315 320 His Arg Gln Arg Arg Ser Thr Lys Asp Asn Phe Asn Val Tyr Lys Ala 325 330 335 Thr Arg Pro Tyr Leu Ala His Cys Pro Asp Cys Gly Glu Gly His Ser 340 345 350 Cys His Ser Pro Ile Ala Leu Glu Arg Ile Arg Asn Glu Ala Thr Asp 355 360 365 Gly Thr Leu Lys Ile Gln Val Ser Leu Gln Ile Gly Ile Lys Thr Asp 370 375 380 Asp Ser His Asp Trp Thr Lys Leu Arg Tyr Met Asp Ser His Thr Pro 385 390 395 400 Ala Asp Ala Glu Arg Ala Gly Leu Leu Val Arg Thr Ser Ala Pro Cys 405 410 415 Thr Ile Thr Gly Thr Met Gly His Phe Ile Leu Ala Arg Cys Pro Lys 420 425 430 Gly Glu Thr Leu Thr Val Gly Phe Thr Asp Ser Arg Lys Ile Ser His 435 440 445 Thr Cys Thr His Pro Phe His His Glu Pro Pro Val Ile Gly Arg Glu 450 455 460 Arg Phe His Ser Arg Pro Gln His Gly Lys Glu Leu Pro Cys Ser Thr 465 470 475 480 Tyr Val Gln Ser Thr Ala Ala Thr Ala Glu Glu Ile Glu Val His Met 485 490 495 Pro Pro Asp Thr Pro Asp Arg Thr Leu Met Thr Gln Gln Ser Gly Asn 500 505 510 Val Lys Ile Thr Val Asn Gly Gln Thr Val Arg Tyr Lys Cys Asn Cys 515 520 525 Gly Gly Ser Gly Gly Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 530 535 540 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Val Asp Pro Asn 545 550 555 560 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 565 570 575 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 580 585 590 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 595 600 605 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 610 615 620 Ala Asn Pro Asn Ala Asn Pro Asn Ala Gly Gly Ser Asn Glu Gly Leu 625 630 635 640 Thr Thr Thr Asp Lys Val Ile Asn Asn Cys Lys Ile Asp Gln Cys His 645 650 655 Ala Ala Val Thr Asn His Lys Asn Trp Gln Tyr Asn Ser Pro Leu Val 660 665 670 Pro Arg Asn Ala Glu Leu Gly Asp Arg Lys Gly Lys Ile His Ile Pro 675 680 685 Phe Pro Leu Ala Asn Val Thr Cys Arg Val Pro Lys Ala Arg Asn Pro 690 695 700 Thr Val Thr Tyr Gly Lys Asn Gln Val Thr Met Leu Leu Tyr Pro Asp 705 710 715 720 His Pro Thr Leu Leu Ser Tyr Arg Asn Met Gly Gln Glu Pro Asn Tyr 725 730 735 His Glu Glu Trp Val Thr His Lys Lys Glu Val Thr Leu Thr Val Pro 740 745 750 Thr Glu Gly Leu Glu Val Thr Trp Gly Asn Asn Glu Pro Tyr Lys Tyr 755 760 765 Trp Pro Gln Met Ser Thr Asn Gly Thr Ala His Gly His Pro His Glu 770 775 780 Ile Ile Leu Tyr Tyr Tyr Glu Leu Tyr Pro Thr Met Thr Val Val Ile 785 790 795 800 Val Ser Val Ala Ser Phe Val Leu Leu Ser Met Val Gly Thr Ala Val 805 810 815 Gly Met Cys Val Cys Ala Arg Arg Arg Cys Ile Thr Pro Tyr Glu Leu 820 825 830 Thr Pro Gly Ala Thr Val Pro Phe Leu Leu Ser Leu Leu Cys Cys Val 835 840 845 Arg Thr Thr Lys Ala Ala Thr Tyr Tyr Glu Ala Ala Ala Tyr Leu Trp 850 855 860 Asn Glu Gln Gln Pro Leu Phe Trp Leu Gln Ala Leu Ile Pro Leu Ala 865 870 875 880 Ala Leu Ile Val Leu Cys Asn Cys Leu Lys Leu Leu Pro Cys Cys Cys 885 890 895 Lys Thr Leu Ala Phe Leu Ala Val Met Ser Ile Gly Ala His Thr Val 900 905 910 Ser Ala Tyr Glu His Val Thr Val Ile Pro Asn Thr Val Gly Val Pro 915 920 925 Tyr Lys Thr Leu Val Asn Arg Pro Gly Tyr Ser Pro Met Val Leu Glu 930 935 940 Met Glu Leu Gln Ser Val Thr Leu Glu Pro Thr Leu Ser Leu Asp Tyr 945 950 955 960 Ile Thr Cys Glu Tyr Lys Thr Val Ile Pro Ser Pro Tyr Val Lys Cys 965 970 975 Cys Gly Thr Ala Glu Cys Lys Asp Lys Ser Leu Pro Asp Tyr Ser Cys 980 985 990 Lys Val Phe Thr Gly Val Tyr Pro Phe Met Trp Gly Gly Ala Tyr Cys 995 1000 1005 Phe Cys Asp Ala Glu Asn Thr Gln Leu Ser Glu Ala His Val Glu 1010 1015 1020 Lys Ser Glu Ser Cys Lys Thr Glu Phe Ala Ser Ala Tyr Arg Ala 1025 1030 1035 His Thr Ala Ser Ala Ser Ala Lys Leu Arg Val Leu Tyr Gln Gly 1040 1045 1050 Asn Asn Ile Thr Val Ala Ala Tyr Ala Asn Gly Asp His Ala Val 1055 1060 1065 Thr Val Lys Asp Ala Lys Phe Val Val Gly Pro Met Ser Ser Ala 1070 1075 1080 Trp Thr Pro Phe Asp Asn Lys Ile Val Val Tyr Lys Gly Asp Val 1085 1090 1095 Tyr Asn Met Asp Tyr Pro Pro Phe Gly Ala Gly Arg Pro Gly Gln 1100 1105 1110 Phe Gly Asp Ile Gln Ser Arg Thr Pro Glu Ser Lys Asp Val Tyr 1115 1120 1125 Ala Asn Thr Gln Leu Val Leu Gln Arg Pro Ala Ala Gly Thr Val 1130 1135 1140 His Val Pro Tyr Ser Gln Ala Pro Ser Gly Phe Lys Tyr Trp Leu 1145 1150 1155 Lys Glu Arg Gly Ala Ser Leu Gln His Thr Ala Pro Phe Gly Cys 1160 1165 1170 Gln Ile Ala Thr Asn Pro Val Arg Ala Val Asn Cys Ala Val Gly 1175 1180 1185 Asn Ile Pro Ile Ser Ile Asp Ile Pro Asp Ala Ala Phe Thr Arg 1190 1195 1200 Val Val Asp Ala Pro Ser Val Thr Asp Met Ser Cys Glu Val Pro 1205 1210 1215 Ala Cys Thr His Ser Ser Asp Phe Gly Gly Val Ala Ile Ile Lys 1220 1225 1230 Tyr Thr Ala Ser Lys Lys Gly Lys Cys Ala Val His Ser Met Thr 1235 1240 1245 Asn Ala Val Thr Ile Arg Glu Ala Asp Val Glu Val Glu Gly Asn 1250 1255 1260 Ser Gln Leu Gln Ile Ser Phe Ser Thr Ala Leu Ala Ser Ala Glu 1265 1270 1275 Phe Arg Val Gln Val Cys Ser Thr Gln Val His Cys Ala Ala Ala 1280 1285 1290 Cys His Pro Pro Lys Asp His Ile Val Asn Tyr Pro Ala Ser His 1295 1300 1305 Thr Thr Leu Gly Val Gln Asp Ile Ser Thr Thr Ala Met Ser Trp 1310 1315 1320 Val Gln Lys Ile Thr Gly Gly Val Gly Leu Ile Val Ala Val Ala 1325 1330 1335 Ala Leu Ile Leu Ile Val Val Leu Cys Val Ser Phe Ser Arg His 1340 1345 1350 427329DNAArtificial SequenceSynthetic VLP74_25 (VEEV VLP 519 NPNAx6) sequence 42tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 240attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 300tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 360tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtgtt ggcattgatt 420attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga 480gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg 540cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg 600acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca 660tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 720ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc 780tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc 840acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 900tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag 960gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agataactgc 1020aggtcgacga tatcgcggcc gccaccatgt tcccgttcca gccaatgtat ccgatgcagc 1080caatgcccta tcgcaacccg ttcgcggccc cgcgcaggcc ctggttcccc agaaccgacc 1140cttttctggc gatgcaggtg caggaattaa cccgctcgat ggctaacctg acgttcaagc 1200aacgccggga cgcgccacct gaggggccat ccgctaataa accgaagaag gaggcctcgc 1260aaaaacagaa agggggaggc caagggaaga agaagaagaa ccaagggaag aagaaggcta 1320agacagggcc gcctaatccg aaggcacaga atggaaacaa gaagaagacc aacaagaaac 1380caggcaagag acagcgcatg gtcatgaaat tggaatctga caagacgttc ccaatcatgt 1440tggaagggaa gataaacggc tacgcttgtg tggtcggagg gaagttattc aggccgatgc 1500atgtggaagg caagatcgac aacgacgttc tggccgcgct taagacgaag aaagcatcca 1560aatacgatct tgagtatgca gatgtgccac agaacatgcg ggccgataca ttcaaataca 1620cccatgagaa accccaaggc tattacagct ggcatcatgg agcagtccaa tatgaaaatg 1680ggcgtttcac ggtgccgaaa ggagttgggg ccaagggaga cagcggacga cccattctgg 1740ataaccaggg acgggtggtc gctattgtgc tgggaggtgt gaatgaagga tctaggacag 1800ccctttcagt cgtcatgtgg aacgagaagg gagttaccgt gaagtatact ccagagaact 1860gcgagcaatg gtcactagtg accaccatgt gtctgctcgc caatgtgacg ttcccatgtg 1920ctcaaccacc aatttgctac gacagaaaac cagcagagac tttggccatg ctcagcgtta 1980acgttgacaa cccgggctac gatgagctgc tggaagcagc tgttaagtgc cccggaagga 2040aaaggagatc caccgaggag ctgtttaatg agtataagct aacgcgccct tacatggcca 2100gatgcatcag atgtgcagtt gggagctgcc atagtccaat agcaatcgag gcagtaaaga 2160gcgacgggca cgacggttat gttagacttc agacttcctc gcagtatggc ctggattcct 2220ccggcaactt aaagggcagg accatgcggt atgacatgca cgggaccatt aaagagatac 2280cactacatca agtgtcactc tatacatctc gcccgtgtca cattgtggat gggcacggtt 2340atttcctgct tgccaggtgc ccggcagggg actccatcac catggaattt aagaaagatt 2400ccgtcagaca ctcctgctcg gtgccgtatg aagtgaaatt taatcctgta ggcagagaac 2460tctatactca tcccccagaa cacggagtag agcaagcgtg ccaagtctac gcacatgatg 2520cacagaacag aggagcttat gtcgagatgc acctcccggg ctcagaagtg gacagcagtt 2580tggtttcctt gagcggcagt tccggaggaa acccgaatgc caatcccaac gcgaacccca 2640atgctaaccc aaatgccaac ccaaacgcca accccaacgc tggtggatcc tcagtcaccg 2700tgacacctcc tgatgggact agcgccctgg tggaatgcga gtgtggcggc acaaagatct 2760ccgagaccat caacaagaca aaacagttca gccagtgcac aaagaaggag cagtgcagag 2820catatcggct gcagaacgat aagtgggtgt ataattctga caaactgccc aaagcagcgg 2880gagccacctt aaaaggaaaa ctgcatgtcc cattcttgct ggcagacggc aaatgcaccg 2940tgcctctagc accagaacct atgataacct tcggtttcag atcagtgtca ctgaaactgc 3000accctaagaa tcccacatat ctaatcaccc gccaacttgc tgatgagcct cactacacgc 3060acgagctcat atctgaacca gctgttagga attttaccgt caccgaaaaa gggtgggagt 3120ttgtatgggg aaaccacccg ccgaaaaggt tttgggcaca ggaaacagca cccggaaatc 3180cacatgggct accgcacgag gtgataactc attattacca cagataccct atgtccacca 3240tcctgggttt gtcaatttgt gccgccattg caaccgtttc cgttgcagcg tctacctggc 3300tgttttgcag atcaagagtt gcgtgcctaa ctccttaccg gctaacacct aacgctagga 3360taccattttg tctggctgtg ctttgctgcg cccgcactgc ccgggccgag accacctggg 3420agtccttgga tcacctatgg aacaataacc aacagatgtt ctggattcaa ttgctgatcc 3480ctctggccgc cttgatcgta gtgactcgcc tgctcaggtg cgtgtgctgt gtcgtgcctt 3540ttttagtcat ggccggcgcc gcaggcgccg gcgcctacga gcacgcgacc acgatgccga 3600gccaagcggg aatctcgtat aacactatag tcaacagagc aggctacgca ccactcccta 3660tcagcataac accaacaaag atcaagctga tacctacagt gaacttggag tacgtcacct 3720gccactacaa aacaggaatg gattcaccag ccatcaaatg ctgcggatct caggaatgca 3780ctccaactta caggcctgat gaacagtgca aagtcttcac aggggtttac ccgttcatgt 3840ggggtggtgc atattgcttt tgcgacactg agaacaccca agtcagcaag gcctacgtaa 3900tgaaatctga cgactgcctt gcggatcatg ctgaagcata taaagcgcac acagcctcag 3960tgcaggcgtt cctcaacatc acagtgggag aacactctat tgtgactacc gtgtatgtga 4020atggagaaac tcctgtgaat ttcaatgggg tcaaaataac tgcaggtccg ctttccacag 4080cttggacacc ctttgatcgc aaaatcgtgc agtatgccgg ggagatatat aattatgatt 4140ttcctgagta tggggcagga caaccaggag catttggaga tatacaatcc agaacagtct 4200caagctctga tctgtatgcc aataccaacc tagtgctgca gagacccaaa gcaggagcga 4260tccacgtgcc atacactcag gcaccttcgg gttttgagca atggaagaaa gataaagctc 4320catcattgaa atttaccgcc cctttcggat gcgaaatata tacaaacccc

attcgcgccg 4380aaaactgtgc tgtagggtca attccattag cctttgacat tcccgacgcc ttgttcacca 4440gggtgtcaga aacaccgaca ctttcagcgg ccgaatgcac tcttaacgag tgcgtgtatt 4500cttccgactt tggtgggatc gccacggtca agtactcggc cagcaagtca ggcaagtgcg 4560cagtccatgt gccatcaggg actgctaccc taaaagaagc agcagtcgag ctaaccgagc 4620aagggtcggc gactatccat ttctcgaccg caaatatcca cccggagttc aggctccaaa 4680tatgcacatc atatgttacg tgcaaaggtg attgtcaccc cccgaaagac catattgtga 4740cacaccctca gtatcacgcc caaacattta cagccgcggt gtcaaaaacc gcgtggacgt 4800ggttaacatc cctgctggga ggatcagccg taattattat aattggcttg gtgctggcta 4860ctattgtggc catgtacgtg ctgaccaacc agaaacataa ttaaggatct agatctgctg 4920tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg 4980aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga 5040gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg 5100aagacaatag caggcatgct ggggatgcgg tgggctctat ggctcgagca tggtcatagc 5160tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 5220taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 5280cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 5340gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 5400tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 5460tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 5520ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 5580agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 5640accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 5700ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 5760gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 5820ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 5880gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 5940taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 6000tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 6060gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 6120cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 6180agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 6240cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 6300cttggtctga cagttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat 6360caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac 6420cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa 6480catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac 6540catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt 6600gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat 6660tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac 6720aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac 6780ctgaatcagg atattcttct aatacctgga atgctgtttt cccagggatc gcagtggtga 6840gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt 6900ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc 6960catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac 7020ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg 7080aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata ctcttccttt 7140ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 7200gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 7260acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc 7320cctttcgtc 7329431285PRTArtificial SequenceSynthetic VLP74_25 (VEEV VLP 519 NPNAx6) sequence 43Met Phe Pro Phe Gln Pro Met Tyr Pro Met Gln Pro Met Pro Tyr Arg 1 5 10 15 Asn Pro Phe Ala Ala Pro Arg Arg Pro Trp Phe Pro Arg Thr Asp Pro 20 25 30 Phe Leu Ala Met Gln Val Gln Glu Leu Thr Arg Ser Met Ala Asn Leu 35 40 45 Thr Phe Lys Gln Arg Arg Asp Ala Pro Pro Glu Gly Pro Ser Ala Asn 50 55 60 Lys Pro Lys Lys Glu Ala Ser Gln Lys Gln Lys Gly Gly Gly Gln Gly 65 70 75 80 Lys Lys Lys Lys Asn Gln Gly Lys Lys Lys Ala Lys Thr Gly Pro Pro 85 90 95 Asn Pro Lys Ala Gln Asn Gly Asn Lys Lys Lys Thr Asn Lys Lys Pro 100 105 110 Gly Lys Arg Gln Arg Met Val Met Lys Leu Glu Ser Asp Lys Thr Phe 115 120 125 Pro Ile Met Leu Glu Gly Lys Ile Asn Gly Tyr Ala Cys Val Val Gly 130 135 140 Gly Lys Leu Phe Arg Pro Met His Val Glu Gly Lys Ile Asp Asn Asp 145 150 155 160 Val Leu Ala Ala Leu Lys Thr Lys Lys Ala Ser Lys Tyr Asp Leu Glu 165 170 175 Tyr Ala Asp Val Pro Gln Asn Met Arg Ala Asp Thr Phe Lys Tyr Thr 180 185 190 His Glu Lys Pro Gln Gly Tyr Tyr Ser Trp His His Gly Ala Val Gln 195 200 205 Tyr Glu Asn Gly Arg Phe Thr Val Pro Lys Gly Val Gly Ala Lys Gly 210 215 220 Asp Ser Gly Arg Pro Ile Leu Asp Asn Gln Gly Arg Val Val Ala Ile 225 230 235 240 Val Leu Gly Gly Val Asn Glu Gly Ser Arg Thr Ala Leu Ser Val Val 245 250 255 Met Trp Asn Glu Lys Gly Val Thr Val Lys Tyr Thr Pro Glu Asn Cys 260 265 270 Glu Gln Trp Ser Leu Val Thr Thr Met Cys Leu Leu Ala Asn Val Thr 275 280 285 Phe Pro Cys Ala Gln Pro Pro Ile Cys Tyr Asp Arg Lys Pro Ala Glu 290 295 300 Thr Leu Ala Met Leu Ser Val Asn Val Asp Asn Pro Gly Tyr Asp Glu 305 310 315 320 Leu Leu Glu Ala Ala Val Lys Cys Pro Gly Arg Lys Arg Arg Ser Thr 325 330 335 Glu Glu Leu Phe Asn Glu Tyr Lys Leu Thr Arg Pro Tyr Met Ala Arg 340 345 350 Cys Ile Arg Cys Ala Val Gly Ser Cys His Ser Pro Ile Ala Ile Glu 355 360 365 Ala Val Lys Ser Asp Gly His Asp Gly Tyr Val Arg Leu Gln Thr Ser 370 375 380 Ser Gln Tyr Gly Leu Asp Ser Ser Gly Asn Leu Lys Gly Arg Thr Met 385 390 395 400 Arg Tyr Asp Met His Gly Thr Ile Lys Glu Ile Pro Leu His Gln Val 405 410 415 Ser Leu Tyr Thr Ser Arg Pro Cys His Ile Val Asp Gly His Gly Tyr 420 425 430 Phe Leu Leu Ala Arg Cys Pro Ala Gly Asp Ser Ile Thr Met Glu Phe 435 440 445 Lys Lys Asp Ser Val Arg His Ser Cys Ser Val Pro Tyr Glu Val Lys 450 455 460 Phe Asn Pro Val Gly Arg Glu Leu Tyr Thr His Pro Pro Glu His Gly 465 470 475 480 Val Glu Gln Ala Cys Gln Val Tyr Ala His Asp Ala Gln Asn Arg Gly 485 490 495 Ala Tyr Val Glu Met His Leu Pro Gly Ser Glu Val Asp Ser Ser Leu 500 505 510 Val Ser Leu Ser Gly Ser Ser Gly Gly Asn Pro Asn Ala Asn Pro Asn 515 520 525 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 530 535 540 Ala Gly Gly Ser Ser Val Thr Val Thr Pro Pro Asp Gly Thr Ser Ala 545 550 555 560 Leu Val Glu Cys Glu Cys Gly Gly Thr Lys Ile Ser Glu Thr Ile Asn 565 570 575 Lys Thr Lys Gln Phe Ser Gln Cys Thr Lys Lys Glu Gln Cys Arg Ala 580 585 590 Tyr Arg Leu Gln Asn Asp Lys Trp Val Tyr Asn Ser Asp Lys Leu Pro 595 600 605 Lys Ala Ala Gly Ala Thr Leu Lys Gly Lys Leu His Val Pro Phe Leu 610 615 620 Leu Ala Asp Gly Lys Cys Thr Val Pro Leu Ala Pro Glu Pro Met Ile 625 630 635 640 Thr Phe Gly Phe Arg Ser Val Ser Leu Lys Leu His Pro Lys Asn Pro 645 650 655 Thr Tyr Leu Ile Thr Arg Gln Leu Ala Asp Glu Pro His Tyr Thr His 660 665 670 Glu Leu Ile Ser Glu Pro Ala Val Arg Asn Phe Thr Val Thr Glu Lys 675 680 685 Gly Trp Glu Phe Val Trp Gly Asn His Pro Pro Lys Arg Phe Trp Ala 690 695 700 Gln Glu Thr Ala Pro Gly Asn Pro His Gly Leu Pro His Glu Val Ile 705 710 715 720 Thr His Tyr Tyr His Arg Tyr Pro Met Ser Thr Ile Leu Gly Leu Ser 725 730 735 Ile Cys Ala Ala Ile Ala Thr Val Ser Val Ala Ala Ser Thr Trp Leu 740 745 750 Phe Cys Arg Ser Arg Val Ala Cys Leu Thr Pro Tyr Arg Leu Thr Pro 755 760 765 Asn Ala Arg Ile Pro Phe Cys Leu Ala Val Leu Cys Cys Ala Arg Thr 770 775 780 Ala Arg Ala Glu Thr Thr Trp Glu Ser Leu Asp His Leu Trp Asn Asn 785 790 795 800 Asn Gln Gln Met Phe Trp Ile Gln Leu Leu Ile Pro Leu Ala Ala Leu 805 810 815 Ile Val Val Thr Arg Leu Leu Arg Cys Val Cys Cys Val Val Pro Phe 820 825 830 Leu Val Met Ala Gly Ala Ala Gly Ala Gly Ala Tyr Glu His Ala Thr 835 840 845 Thr Met Pro Ser Gln Ala Gly Ile Ser Tyr Asn Thr Ile Val Asn Arg 850 855 860 Ala Gly Tyr Ala Pro Leu Pro Ile Ser Ile Thr Pro Thr Lys Ile Lys 865 870 875 880 Leu Ile Pro Thr Val Asn Leu Glu Tyr Val Thr Cys His Tyr Lys Thr 885 890 895 Gly Met Asp Ser Pro Ala Ile Lys Cys Cys Gly Ser Gln Glu Cys Thr 900 905 910 Pro Thr Tyr Arg Pro Asp Glu Gln Cys Lys Val Phe Thr Gly Val Tyr 915 920 925 Pro Phe Met Trp Gly Gly Ala Tyr Cys Phe Cys Asp Thr Glu Asn Thr 930 935 940 Gln Val Ser Lys Ala Tyr Val Met Lys Ser Asp Asp Cys Leu Ala Asp 945 950 955 960 His Ala Glu Ala Tyr Lys Ala His Thr Ala Ser Val Gln Ala Phe Leu 965 970 975 Asn Ile Thr Val Gly Glu His Ser Ile Val Thr Thr Val Tyr Val Asn 980 985 990 Gly Glu Thr Pro Val Asn Phe Asn Gly Val Lys Ile Thr Ala Gly Pro 995 1000 1005 Leu Ser Thr Ala Trp Thr Pro Phe Asp Arg Lys Ile Val Gln Tyr 1010 1015 1020 Ala Gly Glu Ile Tyr Asn Tyr Asp Phe Pro Glu Tyr Gly Ala Gly 1025 1030 1035 Gln Pro Gly Ala Phe Gly Asp Ile Gln Ser Arg Thr Val Ser Ser 1040 1045 1050 Ser Asp Leu Tyr Ala Asn Thr Asn Leu Val Leu Gln Arg Pro Lys 1055 1060 1065 Ala Gly Ala Ile His Val Pro Tyr Thr Gln Ala Pro Ser Gly Phe 1070 1075 1080 Glu Gln Trp Lys Lys Asp Lys Ala Pro Ser Leu Lys Phe Thr Ala 1085 1090 1095 Pro Phe Gly Cys Glu Ile Tyr Thr Asn Pro Ile Arg Ala Glu Asn 1100 1105 1110 Cys Ala Val Gly Ser Ile Pro Leu Ala Phe Asp Ile Pro Asp Ala 1115 1120 1125 Leu Phe Thr Arg Val Ser Glu Thr Pro Thr Leu Ser Ala Ala Glu 1130 1135 1140 Cys Thr Leu Asn Glu Cys Val Tyr Ser Ser Asp Phe Gly Gly Ile 1145 1150 1155 Ala Thr Val Lys Tyr Ser Ala Ser Lys Ser Gly Lys Cys Ala Val 1160 1165 1170 His Val Pro Ser Gly Thr Ala Thr Leu Lys Glu Ala Ala Val Glu 1175 1180 1185 Leu Thr Glu Gln Gly Ser Ala Thr Ile His Phe Ser Thr Ala Asn 1190 1195 1200 Ile His Pro Glu Phe Arg Leu Gln Ile Cys Thr Ser Tyr Val Thr 1205 1210 1215 Cys Lys Gly Asp Cys His Pro Pro Lys Asp His Ile Val Thr His 1220 1225 1230 Pro Gln Tyr His Ala Gln Thr Phe Thr Ala Ala Val Ser Lys Thr 1235 1240 1245 Ala Trp Thr Trp Leu Thr Ser Leu Leu Gly Gly Ser Ala Val Ile 1250 1255 1260 Ile Ile Ile Gly Leu Val Leu Ala Thr Ile Val Ala Met Tyr Val 1265 1270 1275 Leu Thr Asn Gln Lys His Asn 1280 1285 4420PRTPlasmodium falciparum 44Glu Tyr Leu Asn Lys Ile Gln Asn Ser Leu Ser Thr Glu Trp Ser Pro 1 5 10 15 Cys Ser Val Thr 20 4520PRTPlasmodium yoelii 45Tyr Asn Arg Asn Ile Val Asn Arg Leu Leu Gly Asp Ala Leu Asn Gly 1 5 10 15 Pro Glu Glu Lys 20 4630PRTArtificial SequenceSynthetic VLP74 insert 46Ser Gly Gly Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn 1 5 10 15 Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Gly Gly Ser 20 25 30 4790DNAArtificial SequenceSynthetic VLP74 insert 47tccggaggaa acccgaatgc caatcccaac gcgaacccca atgctaaccc aaatgccaac 60ccaaacgcca accccaacgc tggtggatcc 9048106PRTArtificial SequenceSynthetic VLP78 insert 48Ser Gly Gly Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn 1 5 10 15 Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Val Asp Pro Asn Ala Asn 20 25 30 Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn 35 40 45 Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn 50 55 60 Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn 65 70 75 80 Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn 85 90 95 Pro Asn Ala Asn Pro Asn Ala Gly Gly Ser 100 105 49318DNAArtificial SequenceSynthetic VLP74 insert 49tccggaggaa acccgaatgc caatcccaac gcgaacccca acgctaaccc caacgccaat 60ccgaatgcaa acccgaacgt tgacccaaac gccaacccga atgccaatcc caacgcgaac 120cccaatgcta acccaaatgc caacccaaac gccaacccca acgctaatcc aaacgccaac 180cctaacgcca atcccaacgc gaatcctaac gctaatccca acgcaaatcc caatgctaat 240ccgaacgcga accctaatgc aaaccccaac gccaacccga acgctaaccc gaacgctaat 300cccaacgccg gtggatcc 31850452PRTArtificial SequenceSynthetic CHIKV E2-74 fusion protein 50Ser Thr Lys Asp Asn Phe Asn Val Tyr Lys Ala Thr Arg Pro Tyr Leu 1 5 10 15 Ala His Cys Pro Asp Cys Gly Glu Gly His Ser Cys His Ser Pro Ile 20 25 30 Ala Leu Glu Arg Ile Arg Asn Glu Ala Thr Asp Gly Thr Leu Lys Ile 35 40 45 Gln Val Ser Leu Gln Ile Gly Ile Lys Thr Asp Asp Ser His Asp Trp 50 55 60 Thr Lys Leu Arg Tyr Met Asp Ser His Thr Pro Ala Asp Ala Glu Arg 65 70 75 80 Ala Gly Leu Leu Val Arg Thr Ser Ala Pro Cys Thr Ile Thr Gly Thr 85 90 95 Met Gly His Phe Ile Leu Ala Arg Cys Pro Lys Gly Glu Thr Leu Thr 100 105 110 Val Gly Phe Thr Asp Ser Arg Lys Ile Ser His Thr Cys Thr His Pro 115 120 125 Phe His His Glu Pro Pro Val Ile Gly Arg Glu Arg Phe His Ser Arg 130 135 140 Pro Gln His Gly Lys Glu Leu Pro Cys Ser Thr Tyr Val Gln Ser Thr 145 150 155 160 Ala Ala Thr Ala Glu Glu Ile Glu Val His Met Pro Pro Asp Thr Pro 165 170 175 Asp Arg Thr Leu Met Thr Gln Gln Ser Gly Asn Val Lys Ile Thr Val 180 185 190 Asn Gly Gln Thr Val Arg Tyr Lys Cys Asn Cys Gly Gly Ser Gly Gly 195 200 205 Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala 210 215 220 Asn Pro Asn Ala Asn Pro Asn Ala Gly Gly Ser Asn Glu Gly Leu Thr 225 230 235

240 Thr Thr Asp Lys Val Ile Asn Asn Cys Lys Ile Asp Gln Cys His Ala 245 250 255 Ala Val Thr Asn His Lys Asn Trp Gln Tyr Asn Ser Pro Leu Val Pro 260 265 270 Arg Asn Ala Glu Leu Gly Asp Arg Lys Gly Lys Ile His Ile Pro Phe 275 280 285 Pro Leu Ala Asn Val Thr Cys Arg Val Pro Lys Ala Arg Asn Pro Thr 290 295 300 Val Thr Tyr Gly Lys Asn Gln Val Thr Met Leu Leu Tyr Pro Asp His 305 310 315 320 Pro Thr Leu Leu Ser Tyr Arg Asn Met Gly Gln Glu Pro Asn Tyr His 325 330 335 Glu Glu Trp Val Thr His Lys Lys Glu Val Thr Leu Thr Val Pro Thr 340 345 350 Glu Gly Leu Glu Val Thr Trp Gly Asn Asn Glu Pro Tyr Lys Tyr Trp 355 360 365 Pro Gln Met Ser Thr Asn Gly Thr Ala His Gly His Pro His Glu Ile 370 375 380 Ile Leu Tyr Tyr Tyr Glu Leu Tyr Pro Thr Met Thr Val Val Ile Val 385 390 395 400 Ser Val Ala Ser Phe Val Leu Leu Ser Met Val Gly Thr Ala Val Gly 405 410 415 Met Cys Val Cys Ala Arg Arg Arg Cys Ile Thr Pro Tyr Glu Leu Thr 420 425 430 Pro Gly Ala Thr Val Pro Phe Leu Leu Ser Leu Leu Cys Cys Val Arg 435 440 445 Thr Thr Lys Ala 450 51439PRTArtificial SequenceSynthetic CHIKV E1 51Tyr Glu His Val Thr Val Ile Pro Asn Thr Val Gly Val Pro Tyr Lys 1 5 10 15 Thr Leu Val Asn Arg Pro Gly Tyr Ser Pro Met Val Leu Glu Met Glu 20 25 30 Leu Gln Ser Val Thr Leu Glu Pro Thr Leu Ser Leu Asp Tyr Ile Thr 35 40 45 Cys Glu Tyr Lys Thr Val Ile Pro Ser Pro Tyr Val Lys Cys Cys Gly 50 55 60 Thr Ala Glu Cys Lys Asp Lys Ser Leu Pro Asp Tyr Ser Cys Lys Val 65 70 75 80 Phe Thr Gly Val Tyr Pro Phe Met Trp Gly Gly Ala Tyr Cys Phe Cys 85 90 95 Asp Ala Glu Asn Thr Gln Leu Ser Glu Ala His Val Glu Lys Ser Glu 100 105 110 Ser Cys Lys Thr Glu Phe Ala Ser Ala Tyr Arg Ala His Thr Ala Ser 115 120 125 Ala Ser Ala Lys Leu Arg Val Leu Tyr Gln Gly Asn Asn Ile Thr Val 130 135 140 Ala Ala Tyr Ala Asn Gly Asp His Ala Val Thr Val Lys Asp Ala Lys 145 150 155 160 Phe Val Val Gly Pro Met Ser Ser Ala Trp Thr Pro Phe Asp Asn Lys 165 170 175 Ile Val Val Tyr Lys Gly Asp Val Tyr Asn Met Asp Tyr Pro Pro Phe 180 185 190 Gly Ala Gly Arg Pro Gly Gln Phe Gly Asp Ile Gln Ser Arg Thr Pro 195 200 205 Glu Ser Lys Asp Val Tyr Ala Asn Thr Gln Leu Val Leu Gln Arg Pro 210 215 220 Ala Ala Gly Thr Val His Val Pro Tyr Ser Gln Ala Pro Ser Gly Phe 225 230 235 240 Lys Tyr Trp Leu Lys Glu Arg Gly Ala Ser Leu Gln His Thr Ala Pro 245 250 255 Phe Gly Cys Gln Ile Ala Thr Asn Pro Val Arg Ala Val Asn Cys Ala 260 265 270 Val Gly Asn Ile Pro Ile Ser Ile Asp Ile Pro Asp Ala Ala Phe Thr 275 280 285 Arg Val Val Asp Ala Pro Ser Val Thr Asp Met Ser Cys Glu Val Pro 290 295 300 Ala Cys Thr His Ser Ser Asp Phe Gly Gly Val Ala Ile Ile Lys Tyr 305 310 315 320 Thr Ala Ser Lys Lys Gly Lys Cys Ala Val His Ser Met Thr Asn Ala 325 330 335 Val Thr Ile Arg Glu Ala Asp Val Glu Val Glu Gly Asn Ser Gln Leu 340 345 350 Gln Ile Ser Phe Ser Thr Ala Leu Ala Ser Ala Glu Phe Arg Val Gln 355 360 365 Val Cys Ser Thr Gln Val His Cys Ala Ala Ala Cys His Pro Pro Lys 370 375 380 Asp His Ile Val Asn Tyr Pro Ala Ser His Thr Thr Leu Gly Val Gln 385 390 395 400 Asp Ile Ser Thr Thr Ala Met Ser Trp Val Gln Lys Ile Thr Gly Gly 405 410 415 Val Gly Leu Ile Val Ala Val Ala Ala Leu Ile Leu Ile Val Val Leu 420 425 430 Cys Val Ser Phe Ser Arg His 435 52261PRTArtificial SequenceSynthetic CHIKV capsid 52Met Glu Phe Ile Pro Thr Gln Thr Phe Tyr Asn Arg Arg Tyr Gln Pro 1 5 10 15 Arg Pro Trp Ala Pro Arg Pro Thr Ile Gln Val Ile Arg Pro Arg Pro 20 25 30 Arg Pro Gln Arg Gln Ala Gly Gln Leu Ala Gln Leu Ile Ser Ala Val 35 40 45 Asn Lys Leu Thr Met Arg Ala Val Pro Gln Gln Lys Pro Arg Arg Asn 50 55 60 Arg Lys Asn Lys Lys Gln Arg Gln Lys Lys Gln Ala Pro Gln Asn Asp 65 70 75 80 Pro Lys Gln Lys Lys Gln Pro Pro Gln Lys Lys Pro Ala Gln Lys Lys 85 90 95 Lys Lys Pro Gly Arg Arg Glu Arg Met Cys Met Lys Ile Glu Asn Asp 100 105 110 Cys Ile Phe Glu Val Lys His Glu Gly Lys Val Met Gly Tyr Ala Cys 115 120 125 Leu Val Gly Asp Lys Val Met Lys Pro Ala His Val Lys Gly Thr Ile 130 135 140 Asp Asn Ala Asp Leu Ala Lys Leu Ala Phe Lys Arg Ser Ser Lys Tyr 145 150 155 160 Asp Leu Glu Cys Ala Gln Ile Pro Val His Met Lys Ser Asp Ala Ser 165 170 175 Lys Phe Thr His Glu Lys Pro Glu Gly Tyr Tyr Asn Trp His His Gly 180 185 190 Ala Val Gln Tyr Ser Gly Gly Arg Phe Thr Ile Pro Thr Gly Ala Gly 195 200 205 Lys Pro Gly Asp Ser Gly Arg Pro Ile Phe Asp Asn Lys Gly Arg Val 210 215 220 Val Ala Ile Val Leu Gly Gly Ala Asn Glu Gly Ala Arg Thr Ala Leu 225 230 235 240 Ser Val Val Thr Trp Asn Lys Asp Ile Val Thr Lys Ile Thr Pro Glu 245 250 255 Gly Ala Glu Glu Trp 260 53453PRTArtificial SequenceSynthetic VEEV E2-74 fusion protein 53Ser Thr Glu Glu Leu Phe Asn Glu Tyr Lys Leu Thr Arg Pro Tyr Met 1 5 10 15 Ala Arg Cys Ile Arg Cys Ala Val Gly Ser Cys His Ser Pro Ile Ala 20 25 30 Ile Glu Ala Val Lys Ser Asp Gly His Asp Gly Tyr Val Arg Leu Gln 35 40 45 Thr Ser Ser Gln Tyr Gly Leu Asp Ser Ser Gly Asn Leu Lys Gly Arg 50 55 60 Thr Met Arg Tyr Asp Met His Gly Thr Ile Lys Glu Ile Pro Leu His 65 70 75 80 Gln Val Ser Leu Tyr Thr Ser Arg Pro Cys His Ile Val Asp Gly His 85 90 95 Gly Tyr Phe Leu Leu Ala Arg Cys Pro Ala Gly Asp Ser Ile Thr Met 100 105 110 Glu Phe Lys Lys Asp Ser Val Arg His Ser Cys Ser Val Pro Tyr Glu 115 120 125 Val Lys Phe Asn Pro Val Gly Arg Glu Leu Tyr Thr His Pro Pro Glu 130 135 140 His Gly Val Glu Gln Ala Cys Gln Val Tyr Ala His Asp Ala Gln Asn 145 150 155 160 Arg Gly Ala Tyr Val Glu Met His Leu Pro Gly Ser Glu Val Asp Ser 165 170 175 Ser Leu Val Ser Leu Ser Gly Ser Ser Gly Gly Asn Pro Asn Ala Asn 180 185 190 Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn 195 200 205 Pro Asn Ala Gly Gly Ser Ser Val Thr Val Thr Pro Pro Asp Gly Thr 210 215 220 Ser Ala Leu Val Glu Cys Glu Cys Gly Gly Thr Lys Ile Ser Glu Thr 225 230 235 240 Ile Asn Lys Thr Lys Gln Phe Ser Gln Cys Thr Lys Lys Glu Gln Cys 245 250 255 Arg Ala Tyr Arg Leu Gln Asn Asp Lys Trp Val Tyr Asn Ser Asp Lys 260 265 270 Leu Pro Lys Ala Ala Gly Ala Thr Leu Lys Gly Lys Leu His Val Pro 275 280 285 Phe Leu Leu Ala Asp Gly Lys Cys Thr Val Pro Leu Ala Pro Glu Pro 290 295 300 Met Ile Thr Phe Gly Phe Arg Ser Val Ser Leu Lys Leu His Pro Lys 305 310 315 320 Asn Pro Thr Tyr Leu Ile Thr Arg Gln Leu Ala Asp Glu Pro His Tyr 325 330 335 Thr His Glu Leu Ile Ser Glu Pro Ala Val Arg Asn Phe Thr Val Thr 340 345 350 Glu Lys Gly Trp Glu Phe Val Trp Gly Asn His Pro Pro Lys Arg Phe 355 360 365 Trp Ala Gln Glu Thr Ala Pro Gly Asn Pro His Gly Leu Pro His Glu 370 375 380 Val Ile Thr His Tyr Tyr His Arg Tyr Pro Met Ser Thr Ile Leu Gly 385 390 395 400 Leu Ser Ile Cys Ala Ala Ile Ala Thr Val Ser Val Ala Ala Ser Thr 405 410 415 Trp Leu Phe Cys Arg Ser Arg Val Ala Cys Leu Thr Pro Tyr Arg Leu 420 425 430 Thr Pro Asn Ala Arg Ile Pro Phe Cys Leu Ala Val Leu Cys Cys Ala 435 440 445 Arg Thr Ala Arg Ala 450 54442PRTArtificial SequenceSynthetic VEEV E1 54Tyr Glu His Ala Thr Thr Met Pro Ser Gln Ala Gly Ile Ser Tyr Asn 1 5 10 15 Thr Ile Val Asn Arg Ala Gly Tyr Ala Pro Leu Pro Ile Ser Ile Thr 20 25 30 Pro Thr Lys Ile Lys Leu Ile Pro Thr Val Asn Leu Glu Tyr Val Thr 35 40 45 Cys His Tyr Lys Thr Gly Met Asp Ser Pro Ala Ile Lys Cys Cys Gly 50 55 60 Ser Gln Glu Cys Thr Pro Thr Tyr Arg Pro Asp Glu Gln Cys Lys Val 65 70 75 80 Phe Thr Gly Val Tyr Pro Phe Met Trp Gly Gly Ala Tyr Cys Phe Cys 85 90 95 Asp Thr Glu Asn Thr Gln Val Ser Lys Ala Tyr Val Met Lys Ser Asp 100 105 110 Asp Cys Leu Ala Asp His Ala Glu Ala Tyr Lys Ala His Thr Ala Ser 115 120 125 Val Gln Ala Phe Leu Asn Ile Thr Val Gly Glu His Ser Ile Val Thr 130 135 140 Thr Val Tyr Val Asn Gly Glu Thr Pro Val Asn Phe Asn Gly Val Lys 145 150 155 160 Ile Thr Ala Gly Pro Leu Ser Thr Ala Trp Thr Pro Phe Asp Arg Lys 165 170 175 Ile Val Gln Tyr Ala Gly Glu Ile Tyr Asn Tyr Asp Phe Pro Glu Tyr 180 185 190 Gly Ala Gly Gln Pro Gly Ala Phe Gly Asp Ile Gln Ser Arg Thr Val 195 200 205 Ser Ser Ser Asp Leu Tyr Ala Asn Thr Asn Leu Val Leu Gln Arg Pro 210 215 220 Lys Ala Gly Ala Ile His Val Pro Tyr Thr Gln Ala Pro Ser Gly Phe 225 230 235 240 Glu Gln Trp Lys Lys Asp Lys Ala Pro Ser Leu Lys Phe Thr Ala Pro 245 250 255 Phe Gly Cys Glu Ile Tyr Thr Asn Pro Ile Arg Ala Glu Asn Cys Ala 260 265 270 Val Gly Ser Ile Pro Leu Ala Phe Asp Ile Pro Asp Ala Leu Phe Thr 275 280 285 Arg Val Ser Glu Thr Pro Thr Leu Ser Ala Ala Glu Cys Thr Leu Asn 290 295 300 Glu Cys Val Tyr Ser Ser Asp Phe Gly Gly Ile Ala Thr Val Lys Tyr 305 310 315 320 Ser Ala Ser Lys Ser Gly Lys Cys Ala Val His Val Pro Ser Gly Thr 325 330 335 Ala Thr Leu Lys Glu Ala Ala Val Glu Leu Thr Glu Gln Gly Ser Ala 340 345 350 Thr Ile His Phe Ser Thr Ala Asn Ile His Pro Glu Phe Arg Leu Gln 355 360 365 Ile Cys Thr Ser Tyr Val Thr Cys Lys Gly Asp Cys His Pro Pro Lys 370 375 380 Asp His Ile Val Thr His Pro Gln Tyr His Ala Gln Thr Phe Thr Ala 385 390 395 400 Ala Val Ser Lys Thr Ala Trp Thr Trp Leu Thr Ser Leu Leu Gly Gly 405 410 415 Ser Ala Val Ile Ile Ile Ile Gly Leu Val Leu Ala Thr Ile Val Ala 420 425 430 Met Tyr Val Leu Thr Asn Gln Lys His Asn 435 440 55275PRTArtificial SequenceSynthetic VEEV Capsid 55Met Phe Pro Phe Gln Pro Met Tyr Pro Met Gln Pro Met Pro Tyr Arg 1 5 10 15 Asn Pro Phe Ala Ala Pro Arg Arg Pro Trp Phe Pro Arg Thr Asp Pro 20 25 30 Phe Leu Ala Met Gln Val Gln Glu Leu Thr Arg Ser Met Ala Asn Leu 35 40 45 Thr Phe Lys Gln Arg Arg Asp Ala Pro Pro Glu Gly Pro Ser Ala Asn 50 55 60 Lys Pro Lys Lys Glu Ala Ser Gln Lys Gln Lys Gly Gly Gly Gln Gly 65 70 75 80 Lys Lys Lys Lys Asn Gln Gly Lys Lys Lys Ala Lys Thr Gly Pro Pro 85 90 95 Asn Pro Lys Ala Gln Asn Gly Asn Lys Lys Lys Thr Asn Lys Lys Pro 100 105 110 Gly Lys Arg Gln Arg Met Val Met Lys Leu Glu Ser Asp Lys Thr Phe 115 120 125 Pro Ile Met Leu Glu Gly Lys Ile Asn Gly Tyr Ala Cys Val Val Gly 130 135 140 Gly Lys Leu Phe Arg Pro Met His Val Glu Gly Lys Ile Asp Asn Asp 145 150 155 160 Val Leu Ala Ala Leu Lys Thr Lys Lys Ala Ser Lys Tyr Asp Leu Glu 165 170 175 Tyr Ala Asp Val Pro Gln Asn Met Arg Ala Asp Thr Phe Lys Tyr Thr 180 185 190 His Glu Lys Pro Gln Gly Tyr Tyr Ser Trp His His Gly Ala Val Gln 195 200 205 Tyr Glu Asn Gly Arg Phe Thr Val Pro Lys Gly Val Gly Ala Lys Gly 210 215 220 Asp Ser Gly Arg Pro Ile Leu Asp Asn Gln Gly Arg Val Val Ala Ile 225 230 235 240 Val Leu Gly Gly Val Asn Glu Gly Ser Arg Thr Ala Leu Ser Val Val 245 250 255 Met Trp Asn Glu Lys Gly Val Thr Val Lys Tyr Thr Pro Glu Asn Cys 260 265 270 Glu Gln Trp 275 56414PRTPlasmodium falciparum 56Met Met Arg Lys Leu Ala Ile Leu Ser Val Ser Ser Phe Leu Phe Val 1 5 10 15 Glu Ala Leu Phe Gln Glu Tyr Gln Cys Tyr Gly Ser Ser Ser Asn Thr 20 25 30 Arg Val Leu Asn Glu Leu Asn Tyr Asp Asn Ala Gly Thr Asn Leu Tyr 35 40 45 Asn Glu Leu Glu Met Asn Tyr Tyr Gly Lys Gln Glu Asn Trp Tyr Ser 50 55 60 Leu Lys Lys Asn Ser Arg Ser Leu Gly Glu Asn Asp Asp Gly Asn Asn 65 70 75 80 Asn Asn Gly Asp Asn Gly Arg Glu Gly Lys Asp Glu Asp Lys Arg Asp 85 90 95 Gly Asn Asn Glu Asp Asn Glu Lys Leu Arg Lys Pro Lys His Lys Lys 100 105 110 Leu Lys Gln Pro Gly Asp Gly Asn Pro Asp Pro Asn Ala Asn Pro Asn 115 120 125 Val Asp Pro Asn Ala Asn Pro Asn Val Asp Pro Asn Ala Asn Pro Asn 130 135 140 Val Asp Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 145 150 155 160 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 165 170 175 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn

180 185 190 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 195 200 205 Val Asp Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 210 215 220 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 225 230 235 240 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 245 250 255 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 260 265 270 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 275 280 285 Lys Asn Asn Gln Gly Asn Gly Gln Gly His Asn Met Pro Asn Asp Pro 290 295 300 Asn Arg Asn Val Asp Glu Asn Ala Asn Ala Asn Asn Ala Val Lys Asn 305 310 315 320 Asn Asn Asn Glu Glu Pro Ser Asp Lys His Ile Glu Gln Tyr Leu Lys 325 330 335 Lys Ile Lys Asn Ser Ile Ser Thr Glu Trp Ser Pro Cys Ser Val Thr 340 345 350 Cys Gly Asn Gly Ile Gln Val Arg Ile Lys Pro Gly Ser Ala Asn Lys 355 360 365 Pro Lys Asp Glu Leu Asp Tyr Glu Asn Asp Ile Glu Lys Lys Ile Cys 370 375 380 Lys Met Glu Lys Cys Ser Ser Val Phe Asn Val Val Asn Ser Ser Ile 385 390 395 400 Gly Leu Ile Met Val Leu Ser Phe Leu Phe Leu Asn Thr Arg 405 410

Claims

1-44. (canceled)

45. A virus-like particle comprising a virus structural polypeptide and at least one malaria antigen, wherein said virus structural polypeptide is a polypeptide derived from alphavirus or flavivirus.

46. The virus-like particle according to claim 45, wherein said virus structural polypeptide is a polypeptide derived from alphavirus.

47. The virus-like particle according to claim 45, wherein said virus structural polypeptide comprises the capsid and/or the envelope proteins E1 and E2.

48. The virus-like particle according to claim 47, wherein said at least one malaria antigen is inserted into E2 of the envelope protein.

49. The virus-like particle according to claim 45, wherein said virus structural polypeptide is a polypeptide derived from Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV).

50. The virus-like particle according to claim 45, wherein said at least one malaria antigen is a fragment derived from the circumsporozoite protein.

51. The virus-like particle according to claim 45, wherein said at least one malaria antigen is an antigen comprising (NPNA).sub.n wherein n is from 4 to 30 and/or an antigen comprising (EYLNKIQSLSTEWSPCSVT).sub.y wherein y is from 1 to 6.

52. The virus-like particle according to claim 49, wherein said at least one malaria antigen is inserted between 509-510, 510-511, 511-512, 519-520, 529-530, 530-531 or 531-532 of SEQ ID NO: 1 or 2, or between residues 515-516, 516-517, 517-518, 518-519, 519-520, 536-537, 537-538 or 538-539 of SEQ ID NO: 3.

53. The virus-like particle according to claim 49, wherein the particle is Chikungunya virus-like particle consisting of one or more envelope protein E2 into which the antigen is inserted, one or more envelope protein E1 and one or more capsid, and wherein the envelope protein E2 into which the antigen is inserted consists of an amino acid sequence represented by SEQ ID NO: 50; the envelope protein E1 consists of an amino acid sequence represented by SEQ ID NO: 51; and the capsid consists of an amino acid sequence represented by SEQ ID NO: 52.

54. The virus-like particle according to claim 49, wherein the particle is Venezuelan equine encephalitis virus-like particle consisting of one or more envelope protein E2 into which the antigen is inserted, one or more envelope protein E1 and one or more capsid, and wherein the envelope protein E2 into which the antigen is inserted consists of an amino acid sequence represented by SEQ ID NO: 53; the envelope protein E1 consists of an amino acid sequence represented by SEQ ID NO: 54; and the capsid consists of an amino acid sequence represented by SEQ ID NO: 55.

55. An isolated nucleic acid molecule comprising a nucleotide sequence for expressing the particle according to claim 45.

56. A vector comprising the nucleic acid molecule according to claim 55, wherein the vector optionally comprises an expression control sequence operably linked to the nucleic acid molecule.

57. A pharmaceutical composition comprising: (a) the virus-like particle according to claim 45 and/or a nucleic acid molecule comprising a nucleotide sequence for expressing the particle according to claim 45; and (b) a pharmaceutically acceptable carrier.

58. A vaccine composition comprising the virus-like particle according to claim 45.

59. A method of treating malaria in a mammal, comprising administering an effective amount of the virus-like particle according to claim 45 and/or a nucleic acid molecule comprising a nucleotide sequence for expressing the particle according to claim 45 to the mammal in need thereof.

60. A method of treating malaria in a mammal, comprising administering a priming composition comprising at least one first malaria antigen or at least one polynucleotide encoding at least one first malaria antigen; and a boosting composition comprising at least one polypeptide comprising at least one second malaria antigen or at least one polynucleotide encoding at least one second malaria antigen, wherein at least one of said compositions comprises the virus-like particle according to claim 45 and/or a nucleic acid molecule comprising a nucleotide sequence for expressing the particle according to claim 45 to the mammal in need thereof.

61. The method according to claim 60, wherein the polynucleotide encodes substantially all of the circumsporozoite protein.

62. The vaccine composition according to claim 58 for use in the prevention or treatment of malaria, comprising a plurality of malaria-derived antigens.

63. A kit comprising (a) a vaccine composition comprising the virus-like particle according to claim 45; and (b) another vaccine composition comprising the virus-like particle according to claim 45, wherein the particle contained in (a) is a virus-like particle which is different from the particle contained in (b).

64. The kit according to claim 63, further comprising (c) a third vaccine composition comprising a virus-like comprising a virus structural polypeptide and at least one malaria antigen, wherein said virus structural polypeptide is a polypeptide derived from alphavirus or flavivirus, wherein said first vaccine composition is used for priming immunization and said second and third vaccine compositions are used for boosting immunization, and wherein the virus-like particle contained in said third vaccine composition is different from the virus-like particle contained in said first and second vaccine compositions, or is the same as a virus-like particle contained in either said first or second vaccine composition.

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