Screen For Inhibitors Of Filovirus And Uses Therefor

Abstract

The invention provides methods to identify agents useful to prevent, inhibit or treat viral infections, e.g., filovirus infections, as well as compositions having one or more agents to prevent, inhibit or treat viral infection.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation of U.S. patent application Ser. No. 13/127,951, filed Jul. 15, 2011, which claims the benefit of the filing date of U.S. application Ser. No. 61/112,524, filed on Nov. 7, 2008 and U.S. application Ser. No. 61/150,486, filed on Feb. 6, 2009, the disclosures of which are incorporated by reference herein.

BACKGROUND

[0003] Ebolaviruses (family Filoviridae) cause severe hemorrhagic fevers in humans and nonhuman primates, with mortality rates as high as 90% (Sanchez et al., 2007). Ebolaviruses and the closely related Marburgviruses belong to the Filoviridae family (Feldman et al., 2004). Currently, there are no approved vaccines or antivirals for use against filoviruses. Antivirals are not only desirable for local populations in epidemic areas and for health care workers during an outbreak, but also for researchers studying these viruses. Short interfering RNA molecules (Geisbert et al., 2006), and S-adenosylhomocysteine hydrolase inhibitors (Bray et al., 2000; Huggins et al., 1999) have been shown to inhibit Ebola viral growth in vitro and/or in vivo. However, the most effective approach to filovirus control will likely come from a combination of pharmacologic agents with different mechanisms of action (Bray & Paragas, 2002).

[0004] High throughput molecular screening (HTS) is an automated, simultaneous testing of thousands of distinct chemical compounds in models of biological mechanisms or disease. Since authentic Ebolaviruses are biosafety level 4 (BSL-4) agents, HTS with the viruses is not feasible. The lack of sufficient BSL-4 space, trained personnel, and the rigors of working in BSL-4 laboratories have severely hampered basic research with Ebolaviruses as well as the development of vaccines. These limitations have prompted examination of various steps in the Ebolavirus viral life cycle in the absence of infectious virus: (i) replication and transcription were studied by use of reporter gene assays that are based on the expression of necessary viral components from plasmids (Boehmann et al., 2005; Groseth et al., 2005; Muhlberger et al., 1999; Modrof et al., 2003; Modrof et al., 2002); (ii) entry and fusion processes were assessed with pseudotyping assays that rely on the use of recombinant vesicular stomatitis or retroviruses (Yonezawa et al., 2005; Wool-Lewis et al., 1998; Takada et al., 1997; Marzi et al., 2006); and (iii) budding was examined using virus-like particles that are generated from viral proteins provided by protein expression plasmids (Jasenosky et al., 2001; Licata et al., 2004; Noda et al., 2002; McCarthy et al., 2006; Johnson et al., 2006). However, several recent findings suggest that data obtained with these artificial systems may not always be reproducible with live, authentic Ebolavirus (Neumann et al., 2005).

SUMMARY OF THE INVENTION

[0005] The invention provides a method to identify modulators, e.g., inhibitors, of filovirus infection. The method includes contacting a host cell, e.g., a mammalian cell including a human cell or non-human primate cell, with one or more agents and, in one embodiment, a replication incompetent rhabdovirus having filovirus glycoprotein and a mutant rhabdovirus genome with sequences for a reporter gene product. It is then determined whether the one or more agents inhibit the expression or levels of the reporter gene product, e.g., a reporter protein. In one embodiment, at least one agent inhibits reporter expression or levels by at least 50%, 60%, 70% or more, e.g., 80%, 85%, 90% or more, for instance, by at least 95%, that of reporter expression or levels in a corresponding host cell not contacted with the agent(s). In one embodiment, the host cell is contacted with one agent. In one embodiment, the host cell is contacted with a library of agents. For instance, the host cell may be contacted with a chemically synthesized library, cDNA library or siRNA library. The replication incompetent pseudotyped rhabdovirus may be prepared by contacting a host cell with a vector to express mutant rhabdovirus vRNA with a deletion of rhabdovirus glycoprotein sequences and an insertion of reporter gene sequences. Vectors for protein expression include vectors expressing a filovirus glycoprotein and optionally one or more vectors for protein expression of at least one of P, M, N or L rhabdovirus proteins.

[0006] In one embodiment, the invention provides a method to identify one or more agents that inhibit viral infection or replication, e.g., Ebolavirus infection or replication. The method includes contacting a host cell, e.g., a mammalian cell including a human cell or non-human primate cell, with at least one agent and a recombinant negative-sense, single stranded RNA virus, the genome of which contains a deletion of viral sequences, i.e., it is a mutant genome. In one embodiment, the host cell is infected with the virus before being contacted with the one or more agents and in one embodiment, a lysate is prepared, e.g., after contact with the one or more agents. In one embodiment, the deleted viral sequences correspond to those for a viral glycoprotein. In one embodiment, the deleted viral sequences correspond to those for a nonstructural or nonglycosylated viral protein that is essential in trans for viral replication. In one embodiment, the deletion is effective to inhibit or prevent viral replication upon infection of a cell with the recombinant negative-sense, single stranded RNA virus. For example, the deletion may be effective to prevent expression of a functional nonstructural or nonglycosylated protein, or functional glycoprotein, upon infection of a cell with the recombinant negative-sense, single stranded RNA virus. In one embodiment, the deletion may be in filovirus sequences for a viral protein corresponding to Ebola virus VP30. Such a deletion may include a deletion of 1 or more nucleotides, e.g., a deletion of at least 0.1%, 1%, 5%, 10%, 50%, 60%, 70%, 80%, 90%, or any integer in between, and up to 100% of the viral sequences corresponding to those for a nonstructural or nonglycosylated viral protein that is essential in trans for viral replication, e.g., sequences that do not overlap with those for another viral protein encoded by the viral genome. The deletion is one that is stable over multiple passages and is readily detectable, e.g., by RT-PCR. In one embodiment, the deletion may be in rhabdovirus sequences for a rhabdovirus glycoprotein.

[0007] As described herein, a biologically contained Ebolavirus (Ebola.DELTA.VP30) was employed to identify anti-Ebolavirus candidates using a high throughput screening assay. To determine the steps in the viral life cycle inhibited by an anti-viral compound, an Ebolavirus binding/entry assay and a minigenome replication assay were employed. Anti-viral specificity was defined by using viral growth inhibition tests with Ebola.DELTA.VP30, veccinia virus, adenovirus, influenza virus, and vesicular stomatitis virus. Gedunin and gedunin derivatives were identified as anti-Ebolavirus candidates in the high throughput screening assay. These compounds inhibited the growth of Ebola.DELTA.VP30 but not that of vaccinia virus, adenovirus, influenza virus, or vesicular stomatitis virus. Further, these compounds inhibited Ebolavirus binding/entry and some also inhibited viral genome replication and protein expression. Thus, gedunin and gedunin derivatives are potent inhibitors of Ebolavirus in vitro. Their inhibitory mechanisms rely mainly upon virus binding/entry.

[0008] In one embodiment, an isolated recombinant, biologically contained Ebola virus includes a genome which contains a deletion in sequences corresponding to Ebola virus VP30 sequences. The deletion is effective to inhibit or prevent viral replication, e.g., by preventing expression of a functional protein corresponding to Ebola virus VP30 protein, upon infection of a cell that lacks sequences that encode the functional protein (e.g., the cell that does not express functional VP30 in trans) with the recombinant, biologically contained Ebola virus. In one embodiment, at least 90% of sequences corresponding to VP30 sequences in the viral genome of the virus are deleted. In one embodiment, the genome of the recombinant, biologically contained filovirus further comprises heterologous sequences, for instance, positioned within the deletion. The heterologous sequences may be selected as ones that are not toxic to one or more host cells, e.g., reporter, selectable marker or viral sequences (for instance, neoR, a fluorescent protein such as green fluorescent protein (GFP), luciferase or influenza virus sequences for mammalian cells).

[0009] To prepare such virus, a reverse genetics systems for negative-sense RNA viruses was exploited to generate Ebolaviruses that lack the VP30 gene (which encodes an essential transcription factor), termed Ebola.DELTA.VP30 virus. These viruses were maintained, genetically stable, and biologically confined to a cell line expressing VP30. Hence, the Ebola.DELTA.VP30 virus fulfills several criteria of a vaccine virus: it can be grown to reasonably high titers in helper cells, is genetically stable (as determined by sequence analysis after seven serial passages in VP30-expressing Vero cells), and is safe. Moreover, as described herein, the resultant viruses resemble wild-type virus in their life cycle, their morphology, and their growth properties, but could be handled in a non-BSL-4 laboratory, opening new opportunities for study of the Ebolavirus life cycle and for the identification of effective antiviral compounds.

[0010] Other negative-sense, single stranded RNA viruses may likewise be manipulated, e.g., the genome of Nipah virus, Hendravirus, Henipavirus, and the like, may be manipulated to mutate or delete sequences corresponding to those for a nonstructural or nonglycoslyated viral protein that is required for viral replication. Thus, genomes of viruses in the following families may be manipulated to provide for an infectious, biologically contained virus that resembles wild-type virus in its life cycle, morphology, and growth properties, can be grown to reasonably high titers in helper cells, is genetically stable, and is safe: Bornaviridae, Rhabdoviridae, Filoviridae (genera Marburgvirus and Ebolavirus), Paramyxoviridae, Avulavirus, Henipavirus, Morbillivirus, Respirovirus, or Rubulavirus.

[0011] The invention further provides screening methods for antivirals that employ the recombinant infectious, biologically contained virus. In one embodiment, the methods include those that identify one or more agents that inhibit virus infection or replication. The methods include contacting the recombinant infectious, biologically contained virus of the invention, a host cell, e.g., a helper cell, such as a mammalian cell including a human cell or non-human primate cell, and one or more agents. Then it is determined whether the one or more agents inhibit viral replication or infection. In one embodiment, the one or more identified agents do not substantially decrease host cell viability, e.g., host cell viability is at least 65%, 70%, 75%, 80% or more in the presence of the one or more agents. Further provided is a method to identify one or more agents that inhibit virus infection or replication, which includes contacting a host cell infected with a recombinant infectious, biologically contained filovirus, or a lysate thereof, and one or more agents. Then it is determined whether the one or more agents inhibit viral replication or infection. In one embodiment, the one or more identified agents do not substantially decrease host cell viability, e.g., host cell viability is at least 65%, 70%, 75%, 80% or more. In one embodiment, the anti-viral agent has an IC.sub.50 of less than about 10.0 .mu.M, e.g., less than 5 .mu.M, 1 .mu.M, or 0.1 .mu.M, e.g., an IC.sub.50 from 0.001 .mu.M to 10 .mu.M. In one embodiment, the anti-viral agent has a CC.sub.50 of more than about 0.1 .mu.M, e.g., more than 1 .mu.M, 5 .mu.M, 10 .mu.M or 50 .mu.M, e.g., a CC.sub.50 from 0.1 .mu.M to 100 .mu.M. In one embodiment, the agent has an IC.sub.50 of less than about 10.0 .mu.M, e.g., less than 5 .mu.M, 1 .mu.M, or 0.1 .mu.M and a CC.sub.50 of more than about 0.1 .mu.M, e.g., more than 1 .mu.M, 5 .mu.M, 10 .mu.M or 50 .mu.M.

[0012] In one embodiment, the screening method identifies inhibitors of filovirus glycoprotein receptor binding or fusion. The method includes contacting a host cell, e.g., a mammalian cell including a human cell or non-human primate cell, with one or more agents and a recombinant replication incompetent pseudotyped rhabdovirus comprising filovirus glycoprotein and a mutant negative sense rhabdovirus genome which lacks sequences for a rhabdovirus glycoprotein but comprises a sequence for a reporter protein, e.g., a fluorescent protein or a bioluminescent protein. At least one agent is identified that inhibits reporter protein levels or expression in the host cell.

[0013] Also provided is a method which includes contacting a host cell with a plurality of agents, for example, a composition having the plurality of agents, and recombinant virus, e.g., sequentially or simultaneously.

[0014] Further provided are agents identified by the methods and the use of anti-virals in methods to prevent, inhibit or treat viral infection in a mammal, e.g., a human. Agents identified by the method or useful to prevent, inhibit or treat viral, e.g., filovirus, infection, include but are not limited to, an inhibitor of Hsp90, gedunin and gedunin derivatives, a triphenylethylene, an inhibitor of calcium-independent phospholipase A.sub.2 and/or of magnesium-dependent phosphatidate phosphohydrolase, an inhibitor of PGE.sub.2 synthase, a steroid, dopamine antagonist, or anticholinergic, including a compound of formula (I)-(XIII). Such agents are useful treatments in Ebolavirus infection management and biosafety defense, as well as platforms for developing new chemical entities for use in Ebolavirus treatment.

[0015] In addition, the invention provides a method to prevent, inhibit or treat viral infection in a mammal, e.g., a human, by administering a composition having an effective amount of a triphenylethylene, tamoxifen or a derivative thereof such as raloxifene and clomiphene, a calcium channel blocker, a tetranortriterpenoid, an antipsychotic, a sigma receptor agonist, an anticholinergic, a steroid, an inhibitor of calcium-independent phospholipase A.sub.2, an inhibitor of magnesium-dependent phosphatidate phosphohydrolase, an inhibitor of the inducible microsomal PGE.sub.2 synthase, a Hsp90 inhibitor, a dopamine antagonist, or a compound of formula (I)-(XIII), including compositions having those agents or compounds and pharmaceutically acceptable carriers and/or excipients. In one embodiment a composition for administration in prophylactic or therapeutic methods includes but is not limited to bepridil hydrochloride, clomiphene citrate, benzotropin mesylate, 7-deacetoxy-3-deacetyl-7-oxokhivorin, 1,2alpha-epoxy-7-deacetoxy-7-oxodihxdrogedunin, epoxygedunin, 1,3-dideacetyl-7-deacetoxy-7-oxokhivorin, gedunin, gedunol, dihydrogedunin, 3beta-acetoxydeoxodihydrogedunin, 3alpha-hydroxydeoxodihydrogedunin, deacetoxy-7-oxogedunin, 3beta-hydroxydeoxodihydrogedunin, deacetoxy-7-oxogedunin, 1,2alpha-epoxydeacetoxydihydrogedunin, 3beta-hydroxydeoxydesacetoxy-7-oxogedunin, tridesacetoxykhivorin, 1,3-dideacetylkhivorin, heudelottin C, tamoxifen citrate, fluspirilene, raloxifene hydrochloride, bromoenol lactone, cortexolone maleate, (R,R)-cis,diethyl tetrahydro-2,8-chrysenediol, MK-866, L-687, 384 hydrochloride, cycloheximide, HTS00384, NRB03063, CD03565, KM0483, SPB06885, CD04265, CD02075, PD00647, HTS07940, HTS13483, JFD02423, and/or HTS04029.

[0016] Thus, the invention provides compounds for use in medical therapy, such as agents that prevent, inhibit or treat filovirus infection in a mammal, optionally in conjunction with other compounds. Also provided is the use of the compounds for the manufacture of a medicament to prevent, inhibit or treat filovirus infection.

BRIEF DESCRIPTION OF THE FIGURES

[0017] FIGS. 1A-1B. Ebola .DELTA.VP30 virus generates an antibody response against the Ebola virus glycoprotein, GP. (A) Flow chart of vaccination of 4-week-old Balb/c mice with Ebola.DELTA.VP30 virus to determine the antibody titer to Ebola GP. Mice (n=4) were vaccinated three times with 107 FFU of Ebola .DELTA.VP30 at three-week intervals; control mice (n=4) were simultaneously mock-vaccinated. Serum samples were collected two weeks after each vaccination. (B) The amounts of IgG against purified Ebola virus GP in the samples was determined by ELISA. Results are expressed as the mean absorbance at 405 nm (+/-standard deviations) of samples diluted to 1:100.

[0018] FIG. 2. Cellular immune response in Ebola .DELTA.VP30-vaccinated mice. Mice (n=4) were vaccinated with Ebola.DELTA.VP30; control mice (n=2) were simultaneously mock-vaccinated. Splenocytes were collected 8 days after the second vaccination and stimulated with an NP peptide. Cells were stained for the cell surface antigen CD8.sup.+ and for intracellular IFN.gamma.. The number of cytokine-producing CD8.sup.+ T cells was determined by using a FACSCalibur flow i (BD Biosciences).

[0019] FIG. 3. Flow chart of the vaccination schedule to determine the protective efficacy of the Ebola.DELTA.VP30 virus. Four-week-old Balb/c mice were vaccinated with Ebola.DELTA.VP30 virus. In group 1, mice (n=14) were vaccinated with nonpurified Ebola.DELTA.VP30 virus directly from cell culture supernatant, while control mice (n=8) were mock-vaccinated. In group 2, mice (n=15) were vaccinated with purified Ebola.DELTA.VP30 virus, while control mice (n=10) were mock-vaccinated. All mice were challenged with a 1000 MLD.sub.50 of mouse-adapted Ebola virus.

[0020] FIGS. 4A-B. Body weight changes (A) and Kaplan-Meier survival curve (B) of mice vaccinated with Ebola.DELTA.VP30 compared to control mice. Mice from group 1 were vaccinated three times with non-purified Ebola.DELTA.VP30 virus while mice from group 2 were vaccinated twice with purified Ebola.DELTA.VP30 virus. Mice from the vaccinated groups and control groups were challenged with a 1000 MLD.sub.50 of mouse-adapted Ebola virus.

[0021] FIGS. 5A-5B. Ebola .DELTA.VP30 virus generates an antibody response against the Ebola virus glycoprotein, GP. (A) Flow chart of vaccination of 4-week-old Balb/c mice with Ebola.DELTA.VP30 virus to determine the antibody titer to Ebola GP. Mice (n=4) were vaccinated three times with 107 FFU of Ebola .DELTA.VP30 at three-week intervals; control mice (n=4) were simultaneously mock-vaccinated. Serum samples were collected two weeks after each vaccination. (B) The amounts of IgG against purified Ebola virus GP in the samples was determined by ELISA. Results are expressed as the mean absorbance at 405 nm (+/-standard deviations) of samples diluted to 1:100.

[0022] FIG. 6. Cellular immune response in Ebola .DELTA.VP30-vaccinated mice. Mice (n=4) were vaccinated with Ebola.DELTA.VP30; control mice (n=2) were simultaneously mock-vaccinated. Splenocytes were collected 8 days after the second vaccination and stimulated with an NP peptide. Cells were stained for the cell surface antigen CD8.sup.+ and for intracellular IFN.gamma.. The number of cytokine-producing CD8.sup.+ T cells was determined by using a FACSCalibur flow cytometer (BD Biosciences).

[0023] FIG. 7. Flow chart of the vaccination schedule to determine the protective efficacy of the Ebola.DELTA.VP30 virus. Four-week-old Balb/c mice were vaccinated with Ebola.DELTA.VP30 virus. In group 1, mice (n=14) were vaccinated with nonpurified Ebola.DELTA.VP30 virus directly from cell culture supernatant, while control mice (n=8) were mock-vaccinated. In group 2, mice (n=15) were vaccinated with purified Ebola.DELTA.VP30 virus, while control mice (n=10) were mock-vaccinated. All mice were challenged with a 1000 MLD.sub.50 of mouse-adapted Ebola virus.

[0024] FIGS. 8A-B. Body weight changes (A) and Kaplan-Meier survival curve (B) of mice vaccinated with Ebola.DELTA.VP30 compared to control mice. Mice from group 1 were vaccinated three times with non-purified Ebola.DELTA.VP30 virus while mice from group 2 were vaccinated twice with purified Ebola.DELTA.VP30 virus. Mice from the vaccinated groups and control groups were challenged with a 1000 MLD.sub.50 of mouse-adapted Ebola virus.

[0025] FIG. 9. Virus titers in the serum of mice following lethal challenge. Vaccinated (n=3) and control (n=3) mice from groups 1 (top panel) and 2 (lower panel) were euthanized on day 4 post-challenge. Virus titers from the serum were determined by the plaque assay. ND, not detectable.

[0026] FIG. 10A-10BBBBB. Representative filovirus sequences (Accession numbers NC006432, NC004161, AY769362, AY142960, AF522874, AF499101, L11365, NC001608, DQ447652, DQ447649, AB050936, NC002549, NC001608, AF086833 and AF272001, the disclosures of which are incorporated by reference herein; SEQ ID NOs:1-30).

[0027] FIG. 11. Compounds screened in an assay of the invention.

[0028] FIG. 12. Chemical structures of gedunin (1), epoxygedunin (2), 1,3-Dideacetly-7-Deacetoxy-7-Oxokivorin (3), 7-Deacetoxy-3-deacetyl-7-Oxokhivorin (4), and 1,2alpha-Epoxy-7-Deacetoxy-7-Oxo-Deoxyhydrogedunin (5).

[0029] FIG. 13. Growth kinetics of viruses. Compounds were added to cell culture media 2 hours prior to virus infections. Cells were inoculated with Ebola.DELTA.VP30 virus, vaccinia virus, or adenovirus at an MOI of 10.sup.-3, or influenzavirus or VSV at an MOI of 10.sup.-5. Cell culture media (Ebola.DELTA.VP30, influenzavirus, and VSV) or cell culture media and cells (vaccinina virus and adenovirus) were collected 24, 48, and 72 hours post-infection for virus titer determinations. Dots and error bars indicate mean titers and standard deviations from three individual experiments, respectively.

[0030] FIG. 14. Gedunin and gedunin-like compounds inhibit Ebolavirus GP-dependent virus entry. Compounds were added to cell culture media at 2 hours prior to VSV.DELTA.G*-Ebolavirus GP (top panel) or VSV.DELTA.G*-VSV G (lower panel) virus infection. The number of GFP-positive cells was determined after an overnight incubation. % infectivity=100.times.number of GFP-positive cells+compound/number of GFP-positive cells+DMSO. Columns and error bars indicate mean % infectivities and standard deviations from four individual experiments, respectively.

[0031] FIG. 15. Gedunin (1) and epoxygedunin (2) inhibit protein expression from the Ebolavirus minigenome. Compounds were added to cell culture media 6.5 hours post-transfection. Luciferase (luc) activities, expressed from the Ebolavirus minigenome, were measured on day 3 post-transfection. % luc activity=100.times.luc activity+compound/luc activity+DMSO. Columns and error bars indicate mean % luc activities and standard deviations from three individual experiments, respectively.

[0032] FIGS. 16A-16D. Hsp90 inhibitors inhibit protein expression from the Ebolavirus minigenome. (A) Hsp90 inhibitors (10 .mu.M) inhibit growth of Ebola.DELTA.VP30-GFP. Dots and error bars indicate mean titers and standard deviations from three individual experiments, respectively. (B) and (C) Hsp90 inhibitors (10 .mu.M) do not substantially reduce Ebolavirus GP-mediated (or VSV-G-mediated) virus binding/entry. (D) Hsp90 inhibitors (10 .mu.M) reduce protein expression from the Ebolavirus minigenome. Columns and error bars indicate mean % infectivities and standard deviations from four individual experiments, respectively.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

[0033] The term "isolated" when used in relation to a nucleic acid (e.g., vector or plasmid), peptide, polypeptide or virus refers to a nucleic acid sequence, peptide, polypeptide or virus that is identified and separated from at least one contaminant nucleic acid, polypeptide or other biological component with which it is ordinarily associated in its natural source, e.g., so that it is not associated with in vivo substances, or is substantially purified from in vitro substances. Isolated nucleic acid, peptide, polypeptide or virus is present in a form or setting that is different from that in which it is found in nature. For example, a given DNA sequence (e.g., a gene) is found on the host cell chromosome in proximity to neighboring genes; RNA sequences, such as a specific mRNA sequence encoding a specific protein, are found in the cell as a mixture with numerous other mRNAs that encode a multitude of proteins. An example of such DNA "isolated" from a source would be a useful DNA sequence that is excised or removed from said source by chemical means, e.g., by the use of restriction endonucleases, so that it can be further manipulated, e.g., amplified, for use in the invention, by the methodology of genetic engineering. The isolated nucleic acid molecule may be present in single-stranded or double-stranded form. When an isolated nucleic acid molecule is to be utilized to express a protein, the molecule will contain at a minimum the sense or coding strand (i.e., the molecule may single-stranded), but may contain both the sense and anti-sense strands (i.e., the molecule may be double-stranded).

[0034] A "vector" or "construct" (sometimes referred to as gene delivery or gene transfer "vehicle") refers to a macromolecule or complex of molecules comprising a polynucleotide to be delivered to a host cell, either in vitro or in vivo. The polynucleotide to be delivered may comprise a coding sequence of interest for gene therapy. Vectors include, for example, viral vectors (such as adenoviruses, adeno-associated viruses (AAV), lentiviruses, herpesvirus and retroviruses), liposomes and other lipid-containing complexes, and other macromolecular complexes capable of mediating delivery of a polynucleotide to a host cell. Vectors can also comprise other components or functionalities that further modulate gene delivery and/or gene expression, or that otherwise provide beneficial properties to the targeted cells. Such other components include, for example, components that influence binding or targeting to cells (including components that mediate cell-type or tissue-specific binding); components that influence uptake of the vector nucleic acid by the cell; components that influence localization of the polynucleotide within the cell after uptake (such as agents mediating nuclear localization); and components that influence expression of the polynucleotide. Such components also might include markers, such as detectable and/or selectable markers that can be used to detect or select for cells that have taken up and are expressing the nucleic acid delivered by the vector. Such components can be provided as a natural feature of the vector (such as the use of certain viral vectors which have components or functionalities mediating binding and uptake), or vectors can be modified to provide such functionalities. A large variety of such vectors are known in the art and are generally available. When a vector is maintained in a host cell, the vector can either be stably replicated by the cells during mitosis as an autonomous structure, incorporated within the genome of the host cell, or maintained in the host cell's nucleus or cytoplasm.

[0035] A "recombinant viral vector" refers to a viral vector comprising one or more heterologous genes or sequences. Since many viral vectors exhibit size constraints associated with packaging, the heterologous genes or sequences are typically introduced by replacing one or more portions of the viral genome. Such viruses may become replication-defective (biologically contained), requiring the deleted function(s) to be provided in trans during viral replication and encapsidation (by using, e.g., a helper virus or a packaging cell line carrying genes necessary for replication and/or encapsidation). Modified viral vectors in which a polynucleotide to be delivered is carried on the outside of the viral particle have also been described.

[0036] "Gene delivery," "gene transfer," and the like as used herein, are terms referring to the introduction of an exogenous polynucleotide (sometimes referred to as a "transgene") into a host cell, irrespective of the method used for the introduction. Such methods include a variety of well-known techniques such as vector-mediated gene transfer (by, e.g., viral infection/transfection, or various other protein-based or lipid-based gene delivery complexes) as well as techniques facilitating the delivery of "naked" polynucleotides (such as electroporation, "gene gun" delivery and various other techniques used for the introduction of polynucleotides). The introduced polynucleotide may be stably or transiently maintained in the host cell. Stable maintenance typically requires that the introduced polynucleotide either contains an origin of replication compatible with the host cell or integrates into a replicon of the host cell such as an extrachromosomal replicon (e.g., a plasmid) or a nuclear or mitochondrial chromosome. A number of vectors are known to be capable of mediating transfer of genes to mammalian cells, as is known in the art.

[0037] By "transgene" is meant any piece of a nucleic acid molecule (for example, DNA) which is inserted by artifice into a cell either transiently or permanently, and becomes part of the organism if integrated into the genome or maintained extrachromosomally. Such a transgene may include at least a portion of an open reading frame of a gene which is partly or entirely heterologous (i.e., foreign) to the transgenic organism, or may represent at least a portion of an open reading frame of a gene homologous to an endogenous gene of the organism, which portion optionally encodes a polypeptide with substantially the same activity as the corresponding full-length polypeptide or at least one activity of the corresponding full-length polypeptide.

[0038] By "transgenic cell" is meant a cell containing a transgene. For example, a cell stably or transiently transformed with a vector containing an expression cassette is a transgenic cell that can be used to produce a population of cells having altered phenotypic characteristics. A "recombinant cell" is one which has been genetically modified, e.g., by insertion, deletion or replacement of sequences in a nonrecombinant cell by genetic engineering.

[0039] The term "wild-type" or "native" refers to a gene or gene product that has the characteristics of that gene or gene product when isolated from a naturally occurring source. A wild-type gene is that which is most frequently observed in a population and is thus arbitrarily designated the "normal" or "wild-type" form of the gene. In contrast, the term "modified" or "mutant" refers to a gene or gene product that displays modifications in sequence and or functional properties (i.e., altered characteristics) when compared to the wild-type gene or gene product. It is noted that naturally-occurring mutants can be isolated; these are identified by the fact that they have altered characteristics when compared to the wild-type gene or gene product.

[0040] The term "transduction" denotes the delivery of a polynucleotide to a recipient cell either in vivo or in vitro, via a viral vector and in one embodiment via a replication-defective viral vector.

[0041] The term "heterologous" as it relates to nucleic acid sequences such as gene sequences encoding a protein and control sequences, denotes sequences that are not normally joined together, and/or are not normally associated with a particular cell, e.g., are from different sources (for instance, sequences from a virus are heterologous to sequences in the genome of an uninfected cell). Thus, a "heterologous" region of a nucleic acid construct or a vector is a segment of nucleic acid within or attached to another nucleic acid molecule that is not found in association with the other molecule in nature. For example, a heterologous region of a nucleic acid construct could include a coding sequence flanked by sequences not found in association with the coding sequence in nature, i.e., a heterologous promoter. Another example of a heterologous coding sequence is a construct where the coding sequence itself is not found in nature (e.g., synthetic sequences having codons different from the native gene). Similarly, a cell transformed with a construct which is not normally present in the cell would be considered heterologous for purposes of this invention.

[0042] By "DNA" is meant a polymeric form of deoxyribonucleotides (adenine, guanine, thymine, or cytosine) in double-stranded or single-stranded form found, inter alia, in linear DNA molecules (e.g., restriction fragments), viruses, plasmids, and chromosomes. In discussing the structure of particular DNA molecules, sequences may be described herein according to the normal convention of giving only the sequence in the 5' to 3' direction along the nontranscribed strand of DNA (i.e., the strand having the sequence complementary to the mRNA). The term captures molecules that include the four bases adenine, guanine, thymine, or cytosine, as well as molecules that include base analogues which are known in the art.

[0043] As used herein, the terms "complementary" or "complementarity" are used in reference to polynucleotides (i.e., a sequence of nucleotides) related by the base-pairing rules. For example, the sequence "A-G-T," is complementary to the sequence "T-C-A." Complementarity may be "partial," in which only some of the nucleic acids' bases are matched according to the base pairing rules. Or, there may be "complete" or "total" complementarity between the nucleic acids. The degree of complementarity between nucleic acid strands has significant effects on the efficiency and strength of hybridization between nucleic acid strands. This is of particular importance in amplification reactions, as well as detection methods that depend upon binding between nucleic acids.

[0044] DNA molecules are said to have "5' ends" and "3' ends" because mononucleotides are reacted to make oligonucleotides or polynucleotides in a manner such that the 5' phosphate of one mononucleotide pentose ring is attached to the 3' oxygen of its neighbor in one direction via a phosphodiester linkage. Therefore, an end of an oligonucleotide or polynucleotide is referred to as the "5' end" if its 5' phosphate is not linked to the 3' oxygen of a mononucleotide pentose ring and as the "3' end" if its 3' oxygen is not linked to a 5' phosphate of a subsequent mononucleotide pentose ring. As used herein, a nucleic acid sequence, even if internal to a larger oligonucleotide or polynucleotide, also may be said to have 5' and 3' ends. In either a linear or circular DNA molecule, discrete elements are referred to as being "upstream" or 5' of the "downstream" or 3' elements. This terminology reflects the fact that transcription proceeds in a 5' to 3' fashion along the DNA strand. The promoter and enhancer elements that direct transcription of a linked gene are generally located 5' or upstream of the coding region. However, enhancer elements can exert their effect even when located 3' of the promoter element and the coding region. Transcription termination and polyadenylation signals are located 3' or downstream of the coding region.

[0045] A "gene," "polynucleotide," "coding region," "sequence," "segment," "fragment" or "transgene" which "encodes" a particular protein, is a nucleic acid molecule which is transcribed and optionally also translated into a gene product, e.g., a polypeptide, in vitro or in vivo when placed under the control of appropriate regulatory sequences. The coding region may be present in either a cDNA, genomic DNA, or RNA form. When present in a DNA form, the nucleic acid molecule may be single-stranded (i.e., the sense strand) or double-stranded. The boundaries of a coding region are determined by a start codon at 10 the 5' (amino) terminus and a translation stop codon at the 3' (carboxy) terminus. A gene can include, but is not limited to, cDNA from prokaryotic or eukaryotic mRNA, genomic DNA sequences from prokaryotic or eukaryotic DNA, and synthetic DNA sequences. A transcription termination sequence will usually be located 3' to the gene sequence.

[0046] The term "control elements" refers collectively to promoter regions, polyadenylation signals, transcription termination sequences, upstream regulatory domains, origins of replication, internal ribosome entry sites ("IRES"), enhancers, splice junctions, and the like, which collectively provide for the replication, transcription, post-transcriptional processing and translation of a coding sequence in a recipient cell. Not all of these control elements need always be present so long as the selected coding sequence is capable of being replicated, transcribed and translated in an appropriate host cell.

[0047] The term "promoter" is used herein in its ordinary sense to refer to a nucleotide region comprising a DNA regulatory sequence, wherein the regulatory sequence is derived from a gene which is capable of binding RNA polymerase and initiating transcription of a downstream (3' direction) coding sequence.

[0048] By "enhancer" is meant a nucleic acid sequence that, when positioned proximate to a promoter, confers increased transcription activity relative to the transcription activity resulting from the promoter in the absence of the enhancer domain.

[0049] By "operably linked" with reference to nucleic acid molecules is meant that two or more nucleic acid molecules (e.g., a nucleic acid molecule to be transcribed, a promoter, and an enhancer element) are connected in such a way as to permit transcription of the nucleic acid molecule. "Operably linked" with reference to peptide and/or polypeptide molecules is meant that two or more peptide and/or polypeptide molecules are connected in such a way as to yield a single polypeptide chain, i.e., a fusion polypeptide, having at least one property of each peptide and/or polypeptide component of the fusion. The fusion polypeptide is, in one embodiment, chimeric, i.e., composed of heterologous molecules.

[0050] "Homology" refers to the percent of identity between two polynucleotides or two polypeptides. The correspondence between one sequence and to another can be determined by techniques known in the art. For example, homology can be determined by a direct comparison of the sequence information between two polypeptide molecules by aligning the sequence information and using readily available computer programs. Alternatively, homology can be determined by hybridization of polynucleotides under conditions which form stable duplexes between homologous regions, followed by digestion with single strand-specific nuclease(s), and size determination of the digested fragments. Two DNA, or two polypeptide, sequences are "substantially homologous" to each other when at least about 80%, e.g., at least about 90%, or at least about 95% of the nucleotides, or amino acids, respectively match over a defined length of the molecules, as determined using the methods above.

[0051] By "mammal" is meant any member of the class Mammalia including, without limitation, humans and nonhuman primates such as chimpanzees and other apes and monkey species; farm animals such as cattle, sheep, pigs, goats and horses; domestic mammals such as dogs and cats; laboratory animals including rodents such as mice, rats, rabbits and guinea pigs, and the like.

[0052] By "derived from" is meant that a nucleic acid molecule was either made or designed from a parent nucleic acid molecule, the derivative retaining substantially the same functional features of the parent nucleic acid molecule, e.g., encoding a gene product with substantially the same activity as the gene product encoded by the parent nucleic acid molecule from which it was made or designed.

[0053] By "expression construct" or "expression cassette" is meant a nucleic acid molecule that is capable of directing transcription. An expression construct includes, at the least, a promoter. Additional elements, such as an enhancer, and/or a transcription termination signal, may also be included.

[0054] The term "exogenous," when used in relation to a protein, gene, nucleic acid, or polynucleotide in a cell or organism refers to a protein, gene, nucleic acid, or polynucleotide which has been introduced into the cell or organism by artificial or natural means. An exogenous nucleic acid may be from a different organism or cell, or it may be one or more additional copies of a nucleic acid which occurs naturally within the organism or cell. By way of a non-limiting example, an exogenous nucleic acid is in a chromosomal location different from that of natural cells, or is otherwise flanked by a different nucleic acid sequence than that found in nature.

[0055] As used herein, the term "recombinant nucleic acid" or "recombinant DNA sequence, molecule or segment" refers to a nucleic acid, e.g., to DNA, that has been derived or isolated from a source, that may be subsequently chemically altered in vitro, and includes, but is not limited to, a sequence that is naturally occurring, is not naturally occurring, or corresponds to naturally occurring sequences that are not positioned as they would be positioned in the native genome. An example of DNA "derived" from a source, would be a DNA sequence that is identified as a useful fragment, and which is then chemically synthesized in essentially pure form. An example of such DNA "isolated" from a source would be a useful DNA sequence that is excised or removed from said source by chemical means, e.g., by the use of restriction endonucleases, so that it can be further manipulated, e.g., amplified, for use in the invention, by the methodology of genetic engineering.

[0056] The term "recombinant protein" or "recombinant polypeptide" as used herein refers to a protein molecule that is expressed from a recombinant DNA molecule.

[0057] The term "peptide", "polypeptide" and protein" are used interchangeably herein unless otherwise distinguished.

[0058] The term "sequence homology" means the proportion of base matches between two nucleic acid sequences or the proportion amino acid matches between two amino acid sequences. When sequence homology is expressed as a percentage, e.g., 50%, the percentage denotes the proportion of matches over the length of a selected sequence that is compared to some other sequence. Gaps (in either of the two sequences) are permitted to maximize matching; gap lengths of 15 bases or less are usually used, 6 bases or less are preferred with 2 bases or less more preferred. When using oligonucleotides as probes or treatments, the sequence homology between the target nucleic acid and the oligonucleotide sequence is generally not less than 17 target base matches out of 20 possible oligonucleotide base pair matches (85%); such as not less than 9 matches out of 10 possible base pair matches (90%), and, for example, not less than 19 matches out of 20 possible base pair matches (95%).

[0059] The term "selectively hybridize" means to detectably and specifically bind. Polynucleotides, oligonucleotides and fragments of the invention selectively hybridize to nucleic acid strands under hybridization and wash conditions that minimize appreciable amounts of detectable binding to nonspecific nucleic acids. High stringency conditions can be used to achieve selective hybridization conditions as known in the art and discussed herein. Generally, the nucleic acid sequence homology between the polynucleotides, oligonucleotides, and fragments of the invention and a nucleic acid sequence of interest is at least 65%, and more typically with increasing homologies of at least about 70%, about 90%, about 95%, about 98%, and 100%.

[0060] Two amino acid sequences are homologous if there is a partial or complete identity between their sequences. For example, 85% homology means that 85% of the amino acids are identical when the two sequences are aligned for maximum matching. Gaps (in either of the two sequences being matched) are allowed in maximizing matching; gap lengths of 5 or less are preferred with 2 or less being more preferred. Alternatively, two protein sequences (or polypeptide sequences derived from them of at least 30 amino acids in length) are homologous, as this term is used herein, if they have an alignment score of at more than 5 (in standard deviation units) using the program ALIGN with the mutation data matrix and a gap penalty of 6 or greater. The two sequences or parts thereof are more likely homologous if their amino acids are greater than or equal to 50% identical when optimally aligned using the ALIGN program.

[0061] The term "corresponds to" is used herein to mean that a polynucleotide sequence is homologous (e.g., is identical, not strictly evolutionarily related) to all or a portion of a reference polynucleotide sequence that encodes a polypeptide or its complement, or that a polypeptide sequence is identical in sequence or function to a reference polypeptide sequence. For illustration, the nucleotide sequence "TATAC" corresponds to a reference sequence "TATAC" and is complementary to a reference sequence "GTATA".

[0062] The following terms are used to describe the sequence relationships between two or more polynucleotides: "reference sequence", "comparison window", "sequence identity", "percentage of sequence identity", and "substantial identity". A "reference sequence" is a defined sequence used as a basis for a sequence comparison; a reference sequence may be a subset of a larger sequence, for example, as a segment of a full-length cDNA or gene sequence given in a sequence listing, or may comprise a complete cDNA or gene sequence. Generally, a reference sequence is at least 20 nucleotides in length, frequently at least 25 nucleotides in length, and often at least 50 nucleotides in length. Since two polynucleotides may each (1) comprise a sequence (i.e., a portion of the complete polynucleotide sequence) that is similar between the two polynucleotides, and (2) may further comprise a sequence that is divergent between the two polynucleotides, sequence comparisons between two (or more) polynucleotides are typically performed by comparing sequences of the two polynucleotides over a "comparison window" to identify and compare local regions of sequence similarity.

[0063] A "comparison window", as used herein, refers to a conceptual segment of at least 20 contiguous nucleotides and wherein the portion of the polynucleotide sequence in the comparison window may comprise additions or deletions (i.e., gaps) of 20 percent or less as compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences. Optimal alignment of sequences for aligning a comparison window may be conducted by using local homology algorithms or by a search for similarity method, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA Genetics Software Package or by inspection, and the best alignment (i.e., resulting in the highest percentage of homology over the comparison window) generated by the various methods is selected.

[0064] The term "sequence identity" means that two polynucleotide sequences are identical (i.e., on a nucleotide-by-nucleotide basis) over the window of comparison. The term "percentage of sequence identity" means that two polynucleotide sequences are identical (i.e., on a nucleotide-by-nucleotide basis) over the window of comparison. The term "percentage of sequence identity" is calculated by comparing two optimally aligned sequences over the window of comparison, determining the number of positions at which the identical nucleic acid base (e.g., A, T, C, G, U, or I) occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison (i.e., the window size), and multiplying the result by 100 to yield the percentage of sequence identity. The terms "substantial identity" as used herein denote a characteristic of a polynucleotide sequence, wherein the polynucleotide comprises a sequence that has at least 85 percent sequence identity, e.g., at least 90 to 95 percent sequence identity, more usually at least 99 percent sequence identity as compared to a reference sequence over a comparison window of at least 20 nucleotide positions, frequently over a window of at least 20-50 nucleotides, wherein the percentage of sequence identity is calculated by comparing the reference sequence to the polynucleotide sequence which may include deletions or additions which total 20 percent or less of the reference sequence over the window of comparison.

[0065] As applied to polypeptides, the term "substantial identity" means that two peptide sequences, when optimally aligned, such as by the programs GAP or BESTFIT using default gap weights, share at least about 80% sequence identity, e.g., at least about 90% sequence identity, including at least about 95% percent sequence identity, or at least about 99% sequence identity.

[0066] A "protective immune response" and "prophylactic immune response" are used interchangeably to refer to an immune response which targets an immunogen to which the individual has not yet been exposed or targets a protein associated with a disease in an individual who does not have the disease, such as a tumor associated protein in a patient who does not have a tumor.

[0067] A "therapeutic immune response" refers to an immune response which targets an immunogen to which the individual has been exposed or a protein associated with a disease in an individual who has the disease.

[0068] The term "prophylactically effective amount" is meant to refer to the amount necessary to, in the case of infectious agents, prevent an individual from developing an infection, and in the case of diseases, prevent an individual from developing a disease.

[0069] The term "therapeutically effective amount" is meant to refer to the amount necessary to, in the case of infectious agents, reduce the level of infection in an infected individual in order to reduce symptoms or eliminate the infection, and in the case of diseases, to reduce symptoms or cure the individual.

[0070] "Inducing an immune response against an immunogen" is meant to refer to induction of an immune response in a naive individual and induction of an immune response in an individual previously exposed to an immunogen wherein the immune response against the immunogen is enhanced.

[0071] As used herein, "substantially pure" means an object species is the predominant species present (i.e., on a molar basis it is more abundant than any other individual species in the composition), and may be a substantially purified fraction is a composition wherein the object species comprises at least about 50 percent (on a molar basis) of all macromolecular species present. Generally, a substantially pure composition will comprise more than about 80 percent of all macromolecular species present in the composition, for example, more than about 85%, about 90%, about 95%, and about 99%. Min one embodiment, the object species is purified to essential homogeneity (contaminant species cannot be detected in the composition by conventional detection methods) wherein the composition consists essentially of a single macromolecular species.

[0072] "Transfected," "transformed" or "transgenic" is used herein to include any host cell or cell line, which has been altered or augmented by the presence of at least one recombinant DNA sequence. The host cells of the present invention are typically produced by transfection with a DNA sequence in a plasmid expression vector, as an isolated linear DNA sequence, or infection with a recombinant viral vector.

[0073] As used herein, "pharmaceutically acceptable salts" refer to derivatives of the disclosed compounds wherein the parent compound is modified by making acid or base salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. The pharmaceutically acceptable salts include the conventional non-toxic salts or the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, and the like.

[0074] The pharmaceutically acceptable salts of the compounds useful in the present invention can be synthesized from the parent compound, which contains a basic or acidic moiety, by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., p. 1418 (1985), the disclosure of which is hereby incorporated by reference.

[0075] The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit/risk ratio.

[0076] One diastereomer of a compound disclosed herein may display superior activity compared with the other. When required, separation of the racemic material can be achieved by HPLC using a chiral column or by a resolution using a resolving agent such as camphonic chloride as in Thomas J. Tucker, et al., J. Med. Chem. 1994 37, 2437-2444. A chiral compound of Formula I may also be directly synthesized using a chiral catalyst or a chiral ligand, e.g. Mark A. Huffman, et al., J. Org. Chem. 1995, 60, 1590-1594.

[0077] As used herein, "treating" or "treat" includes (i) preventing a pathologic condition from occurring (e.g. prophylaxis); (ii) inhibiting the pathologic condition or arresting its development; (iii) relieving the pathologic condition; and/or diminishing symptoms associated with the pathologic condition.

[0078] As used herein, the term "patient" refers to organisms to be treated by the methods of the present invention. Such organisms include, but are not limited to, mammals such as humans. In the context of the invention, the term "subject" generally refers to an individual who will receive or who has received treatment (e.g., administration of a compound of the invention).

[0079] "Stable compound" and "stable structure" are meant to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent. Only stable compounds are contemplated by the present invention.

[0080] "Substituted" is intended to indicate that one or more hydrogens on the atom indicated in the expression using "substituted" is replaced with a selection from the indicated group(s), provided that the indicated atom's normal valency is not exceeded, and that the substitution results in a stable compound. Suitable indicated groups include, e.g., alkyl, alkenyl, alkylidenyl, alkenylidenyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxycarbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, NR.sup.xR.sup.y and/or COOR.sup.x, wherein each R.sup.x and R.sup.y are independently H, alkyl, alkenyl, aryl, heteroaryl, heterocycle, cycloalkyl or hydroxy. When a substituent is keto (i.e., .dbd.O) or thioxo (i.e., .dbd.S) group, then 2 hydrogens on the atom are replaced.

[0081] "Interrupted" is intended to indicate that in between two or more adjacent carbon atoms, and the hydrogen atoms to which they are attached (e.g., methyl (CH.sub.3), methylene (CH.sub.2) or methine (CH)), indicated in the expression using "interrupted" is inserted with a selection from the indicated group(s), provided that the each of the indicated atoms' normal valency is not exceeded, and that the interruption results in a stable compound. Such suitable indicated groups include, e.g., non-peroxide oxy (--O--), thio (--S--), carbonyl (--C(.dbd.O)--), carboxy (--C(.dbd.O)O--), imine (C.dbd.NH), sulfonyl (SO) or sulfoxide (SO.sub.2).

[0082] Specific and preferred values listed below for radicals, substituents, and ranges, are for illustration only; they do not exclude other defined values or other values within defined ranges for the radicals and substituents

[0083] "Alkyl" refers to a C.sub.1-C.sub.18 hydrocarbon containing normal, secondary, tertiary or cyclic carbon atoms. Examples are methyl (Me, --CH.sub.3), ethyl (Et, --CH.sub.2CH.sub.3), 1-propyl (n-Pr, n-propyl, --CH.sub.2CH.sub.2CH.sub.3), 2-propyl (i-Pr, i-propyl, --CH(CH.sub.3).sub.2), 1-butyl (n-Bu, n-butyl, --CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-methyl-1-propyl (i-Bu, i-butyl, --CH.sub.2CH(CH.sub.3).sub.2), 2-butyl (s-Bu, s-butyl, --CH(CH.sub.3)CH.sub.2CH.sub.3), 2-methyl-2-propyl (t-Bu, t-butyl, --C(CH.sub.3).sub.3), 1-pentyl (n-pentyl, --CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-pentyl (--CH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.3), 3-pentyl (--CH(CH.sub.2CH.sub.3).sub.2), 2-methyl-2-butyl (--C(CH.sub.3).sub.2CH.sub.2CH.sub.3), 3-methyl-2-butyl (--CH(CH.sub.3)CH(CH.sub.3).sub.2), 3-methyl-1-butyl (--CH.sub.2CH.sub.2CH(CH.sub.3).sub.2), 2-methyl-1-butyl (--CH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.3), 1-hexyl (--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-hexyl (--CH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 3-hexyl (--CH(CH.sub.2CH.sub.3)(CH.sub.2CH.sub.2CH.sub.3)), 2-methyl-2-pentyl (--C(CH.sub.3).sub.2CH.sub.2CH.sub.2CH.sub.3), 3-methyl-2-pentyl (--CH(CH.sub.3)CH(CH.sub.3)CH.sub.2CH.sub.3), 4-methyl-2-pentyl (--CH(CH.sub.3)CH.sub.2CH(CH.sub.3).sub.2), 3-methyl-3-pentyl (--C(CH.sub.3)(CH.sub.2CH.sub.3).sub.2), 2-methyl-3-pentyl (--CH(CH.sub.2CH.sub.3)CH(CH.sub.3).sub.2), 2,3-dimethyl-2-butyl (--C(CH.sub.3).sub.2CH(CH.sub.3).sub.2), 3,3-dimethyl-2-butyl (--CH(CH.sub.3)C(CH.sub.3).sub.3.

[0084] The alkyl can optionally be substituted with one or more alkenyl, alkylidenyl, alkenylidenyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxycarbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, NR.sup.xR.sup.y and/or COOR.sup.x, wherein each R.sup.x and R.sup.y are independently H, alkyl, alkenyl, aryl, heteroaryl, heterocycle, cycloalkyl or hydroxyl. The alkyl can optionally be interrupted with one or more non-peroxide oxy (--O--), thio (--S--), carbonyl (--C(.dbd.O)--), carboxy (--C(.dbd.O)O--), sulfonyl (SO) or sulfoxide (SO.sub.2). Additionally, the alkyl can optionally be at least partially unsaturated, thereby providing an alkenyl.

[0085] "Alkenyl" refers to a C.sub.2-C.sub.18 hydrocarbon containing normal, secondary, tertiary or cyclic carbon atoms with at least one site of unsaturation, i.e. a carbon-carbon, sp.sup.2 double bond. Examples include, but are not limited to: ethylene or vinyl (--CH.dbd.CH.sub.2), allyl (--CH.sub.2CH.dbd.CH.sub.2), cyclopentenyl (--C.sub.5H.sub.7), and 5-hexenyl (--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.dbd.CH.sub.2).

[0086] The alkenyl can optionally be substituted with one or more alkyl, alkylidenyl, alkenylidenyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxycarbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, NR.sup.xR.sup.y and/or COOR.sup.x, wherein each R.sup.x and R.sup.y are independently H, alkyl, alkenyl, aryl, heteroaryl, heterocycle, cycloalkyl or hydroxyl. Additionally, the alkenyl can optionally be interrupted with one or more non-peroxide oxy (--O--), thio (--S--), carbonyl (--C(.dbd.O)--), carboxy (--C(.dbd.O)O--), sulfonyl (SO) or sulfoxide (SO.sub.2).

[0087] "Alkylidenyl" refers to a C.sub.1-C.sub.18 hydrocarbon containing normal, secondary, tertiary or cyclic carbon atoms. Examples are methylidenyl (.dbd.CH.sub.2), ethylidenyl (.dbd.CHCH.sub.3), 1-propylidenyl (.dbd.CHCH.sub.2CH.sub.3), 2-propylidenyl (.dbd.C(CH.sub.3).sub.2), 1-butylidenyl (.dbd.CHCH.sub.2CH.sub.2CH.sub.3), 2-methyl-1-propylidenyl (.dbd.CHCH(CH.sub.3).sub.2), 2-butylidenyl (.dbd.C(CH.sub.3)CH.sub.2CH.sub.3), 1-pentyl (.dbd.CHCH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-pentylidenyl (.dbd.C(CH.sub.3)CH.sub.2CH.sub.2CH.sub.3), 3-pentylidenyl (.dbd.C(CH.sub.2CH.sub.3).sub.2), 3-methyl-2-butylidenyl (.dbd.C(CH.sub.3)CH(CH.sub.3).sub.2), 3-methyl-1-butylidenyl (.dbd.CHCH.sub.2CH(CH.sub.3).sub.2), 2-methyl-1-butylidenyl (.dbd.CHCH(CH.sub.3)CH.sub.2CH.sub.3), 1-hexylidenyl (.dbd.CHCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-hexylidenyl (.dbd.C(CH.sub.3)CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 3-hexylidenyl (.dbd.C(CH.sub.2CH.sub.3)(CH.sub.2CH.sub.2CH.sub.3)), 3-methyl-2-pentylidenyl (.dbd.C(CH.sub.3)CH(CH.sub.3)CH.sub.2CH.sub.3), 4-methyl-2-pentylidenyl (.dbd.C(CH.sub.3)CH.sub.2CH(CH.sub.3).sub.2), 2-methyl-3-pentylidenyl (.dbd.C(CH.sub.2CH.sub.3)CH(CH.sub.3).sub.2), and 3,3-dimethyl-2-butylidenyl (.dbd.C(CH.sub.3)C(CH.sub.3).sub.3.

[0088] The alkylidenyl can optionally be substituted with one or more alkyl, alkenyl, alkenylidenyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxycarbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, NR.sup.xR.sup.y and/or COOR.sup.x, wherein each R.sup.x and R.sup.y are independently H, alkyl, alkenyl, aryl, heteroaryl, heterocycle, cycloalkyl or hydroxyl. Additionally, the alkylidenyl can optionally be interrupted with one or more non-peroxide oxy (--O--), thio (--S--), carbonyl (--C(.dbd.O)--), carboxy (--C(.dbd.O)O--), sulfonyl (SO) or sulfoxide (SO.sub.2).

[0089] "Alkenylidenyl" refers to a C.sub.2-C.sub.18 hydrocarbon containing normal, secondary, tertiary or cyclic carbon atoms with at least one site of unsaturation, i.e. a carbon-carbon, sp.sup.2 double bond. Examples include, but are not limited to: allylidenyl (.dbd.CHCH.dbd.CH.sub.2), and 5-hexenylidenyl (.dbd.CHCH.sub.2CH.sub.2CH.sub.2CH.dbd.CH.sub.2).

[0090] The alkenylidenyl can optionally be substituted with one or more alkyl, alkenyl, alkylidenyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxycarbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, NR.sup.xR.sup.y and/or COOR.sup.x, wherein each R.sup.x and R.sup.y are independently H, alkyl, alkenyl, aryl, heteroaryl, heterocycle, cycloalkyl or hydroxyl. Additionally, the alkenylidenyl can optionally be interrupted with one or more non-peroxide oxy (--O--), thio (--S--), carbonyl (--C(.dbd.O)--), carboxy (--C(.dbd.O)O--), sulfonyl (SO) or sulfoxide (SO.sub.2).

[0091] "Alkylene" refers to a saturated, branched or straight chain or cyclic hydrocarbon radical of 1-18 carbon atoms, and having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or different carbon atoms of a parent alkane. Typical alkylene radicals include, but are not limited to: methylene (--CH.sub.2--) 1,2-ethyl (--CH.sub.2CH.sub.2--), 1,3-propyl (--CH.sub.2CH.sub.2CH.sub.2--), 1,4-butyl (--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--), and the like.

[0092] The alkylene can optionally be substituted with one or more alkyl, alkenyl, alkylidenyl, alkenylidenyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxycarbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, NR.sup.xR.sup.y and/or COOR.sup.x, wherein each R.sup.x and R.sup.y are independently H, alkyl, alkenyl, aryl, heteroaryl, heterocycle, cycloalkyl or hydroxyl. Additionally, the alkylene can optionally be interrupted with one or more non-peroxide oxy (--O--), thio (--S--), carbonyl (--C(.dbd.O)--), carboxy (--C(.dbd.O)O--), sulfonyl (SO) or sulfoxide (SO.sub.2). Moreover, the alkylene can optionally be at least partially unsaturated, thereby providing an alkenylene.

[0093] "Alkenylene" refers to an unsaturated, branched or straight chain or cyclic hydrocarbon radical of 2-18 carbon atoms, and having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkene. Typical alkenylene radicals include, but are not limited to: 1,2-ethylene (--CH.dbd.CH--).

[0094] The alkenylene can optionally be substituted with one or more alkyl, alkenyl, alkylidenyl, alkenylidenyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxycarbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, NR.sup.xR.sup.y and/or COOR.sup.x, wherein each R.sup.x and R.sup.y are independently H, alkyl, alkenyl, aryl, heteroaryl, heterocycle, cycloalkyl or hydroxyl. Additionally, The alkenylene can optionally be interrupted with one or more non-peroxide oxy (--O--), thio (--S--), carbonyl (--C(.dbd.O)--), carboxy (--C(.dbd.O)O--), sulfonyl (SO) or sulfoxide (SO.sub.2).

[0095] The term "alkoxy" refers to the groups alkyl-O--, where alkyl is defined herein. Preferred alkoxy groups include, e.g., methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.

[0096] The alkoxy can optionally be substituted with one or more alkyl, alkylidenyl, alkenylidenyl, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxycarbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, NR.sup.xR.sup.y and COOR.sup.x, wherein each R.sup.x and R.sup.y are independently H, alkyl, aryl, heteroaryl, heterocycle, cycloalkyl or hydroxyl.

[0097] The term "aryl" refers to an unsaturated aromatic carbocyclic group of from 6 to 20 carbon atoms having a single ring (e.g., phenyl) or multiple condensed (fused) rings, wherein at least one ring is aromatic (e.g., naphthyl, dihydrophenanthrenyl, fluorenyl, or anthryl). Preferred aryls include phenyl, naphthyl and the like.

[0098] The aryl can optionally be substituted with one or more alkyl, alkenyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxycarbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, NR.sup.xR.sup.y and COOR.sup.x, wherein each R.sup.x and R.sup.y are independently H, alkyl, aryl, heteroaryl, heterocycle, cycloalkyl or hydroxyl.

[0099] The term "cycloalkyl" refers to cyclic alkyl groups of from 3 to 20 carbon atoms having a single cyclic ring or multiple condensed rings. Such cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like, or multiple ring structures such as adamantanyl, and the like.

[0100] The cycloalkyl can optionally be substituted with one or more alkyl, alkenyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heteroaryl, heterocycle, alkanoyl, alkoxycarbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, NR.sup.xR.sup.y and COOR.sup.x, wherein each R.sup.x and R.sup.y are independently H, alkyl, aryl, heteroaryl, heterocycle, cycloalkyl or hydroxyl.

[0101] The cycloalkyl can optionally be at least partially unsaturated, thereby providing a cycloalkenyl.

[0102] The term "halo" refers to fluoro, chloro, bromo, and iodo. Similarly, the term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0103] "Haloalkyl" refers to alkyl as defined herein substituted by 1-4 halo groups as defined herein, which may be the same or different. Representative haloalkyl groups include, by way of example, trifluoromethyl, 3-fluorododecyl, 12,12,12-trifluorododecyl, 2-bromooctyl, 3-bromo-6-chloroheptyl, and the like.

[0104] The term "heteroaryl" is defined herein as a monocyclic, bicyclic, or tricyclic ring system containing one, two, or three aromatic rings and containing at least one nitrogen, oxygen, or sulfur atom in an aromatic ring, and which can be unsubstituted or substituted, for example, with one or more, and in particular one to three, substituents, like halo, alkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, haloalkyl, nitro, amino, alkylamino, acylamino, alkylthio, alkylsulfinyl, and alkylsulfonyl. Examples of heteroaryl groups include, but are not limited to, 2H-pyrrolyl, 3H-indolyl, 4H-quinolizinyl, 4nH-carbazolyl, acridinyl, benzo[b]thienyl, benzothiazolyl, .beta.-carbolinyl, carbazolyl, chromenyl, cinnaolinyl, dibenzo[b,d]furanyl, furazanyl, furyl, imidazolyl, imidizolyl, indazolyl, indolisinyl, indolyl, isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl, naphthyridinyl, naptho[2,3-b], oxazolyl, perimidinyl, phenanthridinyl, phenanthrolinyl, phenarsazinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolyl, quinoxalinyl, thiadiazolyl, thianthrenyl, thiazolyl, thienyl, triazolyl, and xanthenyl. In one embodiment the term "heteroaryl" denotes a monocyclic aromatic ring containing five or six ring atoms containing carbon and 1, 2, 3, or 4 heteroatoms independently selected from the group non-peroxide oxygen, sulfur, and N(Z) wherein Z is absent or is H, O, alkyl, phenyl or benzyl. In another embodiment heteroaryl denotes an ortho-fused bicyclic heterocycle of about eight to ten ring atoms derived therefrom, particularly a benz-derivative or one derived by fusing a propylene, or tetramethylene diradical thereto.

[0105] The heteroaryl can optionally be substituted with one or more alkyl, alkenyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heterocycle, cycloalkyl, alkanoyl, alkoxycarbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, NR.sup.xR.sup.y and COOR.sup.x, wherein each R.sup.x and R.sup.y are independently H, alkyl, aryl, heteroaryl, heterocycle, cycloalkyl or hydroxyl.

[0106] The term "heterocycle" refers to a saturated or partially unsaturated ring system, containing at least one heteroatom selected from the group oxygen, nitrogen, and sulfur, and optionally substituted with alkyl or C(.dbd.O)OR.sup.b, wherein R.sup.b is hydrogen or alkyl. Typically heterocycle is a monocyclic, bicyclic, or tricyclic group containing one or more heteroatoms selected from the group oxygen, nitrogen, and sulfur. A heterocycle group also can contain an oxo group (.dbd.O) attached to the ring. Non-limiting examples of heterocycle groups include 1,3-dihydrobenzofuran, 1,3-dioxolane, 1,4-dioxane, 1,4-dithiane, 2H-pyran, 2-pyrazoline, 4H-pyran, chromanyl, imidazolidinyl, imidazolinyl, indolinyl, isochromanyl, isoindolinyl, morpholine, piperazinyl, piperidine, piperidyl, pyrazolidine, pyrazolidinyl, pyrazolinyl, pyrrolidine, pyrroline, quinuclidine, and thiomorpholine.

[0107] The heterocycle can optionally be substituted with one or more alkyl, alkenyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heteroaryl, cycloalkyl, alkanoyl, alkoxycarbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, NR.sup.xR.sup.y and COOR.sup.x, wherein each R.sup.x and R.sup.y are independently H, alkyl, aryl, heteroaryl, heterocycle, cycloalkyl or hydroxyl.

[0108] Examples of nitrogen heterocycles and heteroaryls include, but are not limited to, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, indoline, morpholino, piperidinyl, tetrahydrofuranyl, and the like as well as N-alkoxy-nitrogen containing heterocycles. In one specific embodiment of the invention, the nitrogen heterocycle can be 3-methyl-5,6-dihydro-4H-pyrazino[3,2,1-jk]carbazol-3-ium iodide.

[0109] Another class of heterocyclics is known as "crown compounds" which refers to a specific class of heterocyclic compounds having one or more repeating units of the formula [--(CH.sub.2-).sub.aA-] where a is equal to or greater than 2, and A at each separate occurrence can be O, N, S or P. Examples of crown compounds include, by way of example only, [--(CH.sub.2).sub.3--NH-].sub.3, [--((CH.sub.2).sub.2--O).sub.4--((CH.sub.2).sub.2--NH).sub.2] and the like. Typically such crown compounds can have from 4 to 10 heteroatoms and 8 to 40 carbon atoms.

[0110] The term "alkanoyl" refers to C(.dbd.O)R, wherein R is an alkyl group as previously defined.

[0111] The term "acyloxy" refers to --O--C(.dbd.O)R, wherein R is an alkyl group as previously defined. Examples of acyloxy groups include, but are not limited to, acetoxy, propanoyloxy, butanoyloxy, and pentanoyloxy. Any alkyl group as defined above can be used to form an acyloxy group.

[0112] The term "alkoxycarbonyl" refers to C(.dbd.O)OR, wherein R is an alkyl group as previously defined.

[0113] The term "amino" refers to --NH.sub.2, and the term "alkylamino" refers to --NR.sub.2, wherein at least one R is alkyl and the second R is alkyl or hydrogen. The term "acylamino" refers to RC(.dbd.O)N, wherein R is alkyl or aryl.

[0114] The term "imino" refers to --C.dbd.NH.

[0115] The term "nitro" refers to --NO.sub.2.

[0116] The term "trifluoromethyl" refers to --CF.sub.3.

[0117] The term "trifluoromethoxy" refers to --OCF.sub.3.

[0118] The term "cyano" refers to --CN.

[0119] The term "hydroxy" or "hydroxyl" refers to --OH.

[0120] The term "oxy" refers to --O--.

[0121] The term "thio" refers to --S--.

[0122] The term "thioxo" refers to (.dbd.S).

[0123] The term "keto" refers to (.dbd.O).

[0124] As to any of the above groups, which contain one or more substituents, it is understood, of course, that such groups do not contain any substitution or substitution patterns which are sterically impractical and/or synthetically non-feasible. In addition, the compounds of this invention include all stereochemical isomers arising from the substitution of these compounds.

[0125] Selected substituents within the compounds described herein are present to a recursive degree. In this context, "recursive substituent" means that a substituent may recite another instance of itself. Because of the recursive nature of such substituents, theoretically, a large number may be present in any given claim. One of ordinary skill in the art of medicinal chemistry understands that the total number of such substituents is reasonably limited by the desired properties of the compound intended. Such properties include, by of example and not limitation, physical properties such as molecular weight, solubility or log P, application properties such as activity against the intended target, and practical properties such as ease of synthesis.

[0126] Recursive substituents are an intended aspect of the invention. One of ordinary skill in the art of medicinal and organic chemistry understands the versatility of such substituents. To the degree that recursive substituents are present in an claim of the invention, the total number will be determined as set forth above.

[0127] The compounds described herein can be administered as the parent compound, a pro-drug of the parent compound, or an active metabolite of the parent compound.

[0128] "Pro-drugs" are intended to include any covalently bonded substances which release the active parent drug or other formulas or compounds of the present invention in vivo when such pro-drug is administered to a mammalian subject. Pro-drugs of a compound of the present invention are prepared by modifying functional groups present in the compound in such a way that the modifications are cleaved, either in routine manipulation in vivo, to the parent compound. Pro-drugs include compounds of the present invention wherein a carbonyl, carboxylic acid, hydroxy or amino group is bonded to any group that, when the pro-drug is administered to a mammalian subject, cleaves to form a free carbonyl, carboxylic acid, hydroxy or amino group. Examples of pro-drugs include, but are not limited to, acetate, formate and benzoate derivatives of alcohol and amine functional groups in the compounds of the present invention, and the like.

[0129] "Metabolite" refers to any substance resulting from biochemical processes by which living cells interact with the active parent drug or other formulas or compounds of the present invention in vivo, when such active parent drug or other formulas or compounds of the present are administered to a mammalian subject. Metabolites include products or intermediates from any metabolic pathway.

[0130] "Metabolic pathway" refers to a sequence of enzyme-mediated reactions that transform one compound to another and provide intermediates and energy for cellular functions. The metabolic pathway can be linear or cyclic.

[0131] Obviously, numerous modifications and variations of the present invention are possible in light of the above teachings. It is therefore to be understood that within the scope of the appended claims, the invention may be practiced otherwise than as specifically described herein.

Methods of the Invention

[0132] The present invention provides a method for screening for compounds that prevent or inhibit viral infection, e.g., prevent or inhibit viral binding to a host cell surface molecule, e.g., receptor, or prevent or inhibit viral membrane fusion with host cell membrane(s). In one embodiment, the screening method includes contacting cells permissive for viral infection with one or more test agents and a recombinant virus, e.g., a pseudotyped virus or a replication defective, e.g., biologically contained, virus to identify agents that prevent or inhibit viral infection. In one embodiment, cells are first contacted with one or more test agents and then with a recombinant virus to identify agents that prevent or inhibit viral infection. In one embodiment, cells are contacted with a recombinant virus and then with one or more test agents. The methods thus identify compounds that may be used alone or in conjunction with other anti-virals, or other prophylactic or therapeutic compounds.

[0133] Agents identified as having anti-viral properties, e.g., agents identified in the screening methods of the invention as having anti-viral properties, are useful in methods to prevent, inhibit or treat viral infection in a mammal. For example, a dopamine antagonist identified as useful to inhibit viral infection or replication in vitro may be employed to prevent, inhibit or treat viral infection in vivo.

Exemplary Viruses Useful in Methods of the Invention

[0134] The invention provides isolated vectors, e.g., plasmids, which encode proteins of negative-sense, single stranded RNA viruses and/or express vRNA from recombinant nucleic acid corresponding to sequences for mutant negative-sense, single stranded RNA viruses. In one embodiment, when introduced into a cell, a combination of these vectors is capable of yielding recombinant infectious, biologically contained virus. Thus, the invention includes host cells that produce recombinant infectious, biologically contained virus. In one embodiment, the invention provides isolated vectors, e.g., plasmids, which encode filovirus proteins and/or express mutant filovirus vRNA which, when introduced into a cell, are capable of yielding recombinant infectious, biologically contained filovirus. In one embodiment, the invention provides isolated vectors, e.g., plasmids, which express mutant negative sense vRNA having reporter sequences and lacking viral glycoprotein sequence and vectors that encode filovirus glycoprotein and optionally non-filovirus proteins which, when introduced into a cell, are capable of yielding a pseudotyped recombinant virus. The invention includes host cells that transiently or stably produce the recombinant virus, including helper cells, and isolated recombinant virus prepared by the methods disclosed herein.

[0135] Thus, vectors of the invention include those for mRNA production and vRNA production. In one embodiment, the vectors include filovirus DNA, for example, vectors for mRNA production with sequences corresponding to one or more open reading frames encoding filovirus proteins, or vectors for vRNA production that include a deletion of the full-length genomic sequence, which deletion includes internal filovirus sequences corresponding to at least a portion of one open reading frame. The RNA produced from the vRNA vector is capable of being packaged into virions in the presence of filovirus proteins but as part of the resulting virion, is not capable of being replicated and so does not result in virus production when that virion is introduced to a cell that otherwise supports filovirus replication and which cell does not express at least one filovirus protein in trans, e.g., a cell that is not a filovirus helper cell.

[0136] For example, Ebolaviruses possess a negative-sense, nonsegmented RNA genome, approximately 19 kilobases in length that encodes seven structural proteins and at least one nonstructural protein (Sanchez et al., 2007). NP, viral protein (VP)35, VP30, and L, the RNA-dependent RNA polymerase, are components of the nucleocapsid involved in viral replication and transcription (Muhlberger et al., 1999). VP40 is the matrix protein and is involved in viral budding (Harty et al., 2000; Panchal et al., 2003). VP24 is involved in the formation of nucleocapsids composed of NP, VP35 and viral RNA (Huang et al., 2002). The only viral surface glycoprotein, GP, plays a role in viral attachment and entry (Chan et al., 2001; Manicassamy et al., 2005; Shimojima et al., 2006; Chandran et al., 2005). Candidate sequences for deletion/mutation and optional replacement with heterologous sequences include but are not limited to Ebola virus VP30 sequences or corresponding sequences in other negative-sense, single stranded RNA viruses, e.g., sequences for nonstructural, nonpolymerase and/or nonglycosylated viral proteins. Although deletions in other Ebola virus sequences, i.e., in GP and VP40, were prepared, only deletions in VP30 sequences resulted in virus that could be recovered. However, deletions in sequences that do not correspond to VP30 sequences in other negative-sense, single stranded RNA viruses may yield infectious, biologically contained virus that is useful in vaccines or in drug screening.

[0137] The vectors may include gene(s) or portions thereof other than those of a negative-sense, single stranded RNA virus such as a filovirus (heterologous sequences), which genes or portions thereof are intended to be expressed in a host cell, either as a protein or incorporated into vRNA. Thus, a vector of the invention may include in addition to viral sequences, for instance, filovirus sequences, a gene or open reading frame of interest, e.g., a heterologous gene for an immunogenic peptide or protein useful as a vaccine or a therapeutic protein.

[0138] To express vRNA, e.g., mutant vRNA, the promoter which is operably linked to viral and reporter gene sequences, which may be in antisense (antigenomic orientation for negative-sense viruses), may be, for example, a RNA polymerase I promoter, a RNA polymerase II promoter, a RNA polymerase III promoter, a T7 promoter, or a T3 promoter. The transcription termination sequence may be a RNA polymerase I transcription termination sequence, a RNA polymerase II transcription termination sequence, a RNA polymerase III transcription termination sequence, or a ribozyme.

[0139] Any promoter may be employed to express a viral protein. A promoter for the vectors includes but is not limited to a RNA polymerase I promoter, a RNA polymerase II promoter, a RNA polymerase III promoter, a T7 promoter, and a T3 promoter. Each vector comprising an open reading frame may include a transcription termination sequence such as a RNA polymerase I transcription termination sequence, a RNA polymerase II transcription termination sequence, a RNA polymerase III transcription termination sequence, or a ribozyme. Preferred promoters for the vectors for vRNA include, but are not limited to, a RNA polymerase I promoter, a RNA polymerase II promoter, a RNA polymerase III promoter, a T7 promoter, and a T3 promoter. In one embodiment, the vector or plasmid which expresses vRNA comprises a promoter, e.g., a RNA polymerase I, suitable for expression in a particular host cell, e.g., avian or mammalian host cells such as canine, feline, equine, bovine, ovine, or primate cells including human cells. In one embodiment, the RNA polymerase I promoter is a human RNA polymerase I promoter. The vectors or plasmids comprising DNA useful to prepare influenza vRNA may comprise RNA polymerase I transcription termination sequences. Preferred transcription termination sequences for the vectors for vRNA include, but are not limited to, a RNA polymerase I transcription termination sequence, a RNA polymerase II transcription termination sequence, or a RNA polymerase III transcription termination sequence, or a ribozyme.

[0140] If more than one vector is employed, the vectors may be physically linked or each vector may be present on an individual plasmid or other, e.g., linear, nucleic acid delivery vehicle. The vectors or plasmids may be introduced to any host cell, e.g., a eukaryotic cell such as a mammalian cell, that supports viral replication. Host cells useful to prepare virus of the invention include but are not limited to insect, avian or mammalian host cells such as canine, feline, equine, bovine, ovine, or primate cells including simian or human cells. In one embodiment, the host cell is one that is approved for vaccine production.

[0141] The viruses produced by methods described herein are useful in viral mutagenesis studies, drug screening and in the production of vaccines (e.g., for AIDS, influenza, hepatitis B, hepatitis C, rhinovirus, filoviruses, malaria, herpes, and foot and mouth disease) and gene therapy vectors (e.g., for cancer, AIDS, adenosine deaminase, muscular dystrophy, ornithine transcarbamylase deficiency and central nervous system tumors). In particular, infectious, biologically contained filovirus of the invention which induces strong humoral and cellular immunity may be employed as a vaccine vector, as they are unlikely to give rise to infectious recombinant virus.

[0142] Thus, a virus for use in medical therapy (e.g., for a vaccine or gene therapy) is provided. For example, the invention provides a method to immunize an animal against a pathogen, e.g., a bacteria, virus such as Ebola virus, or parasite, or a malignant tumor. The method comprises administering to the animal an effective amount of at least one isolated virus of the invention which encodes and expresses, or comprises nucleic acid for an immunogenic peptide or protein of a pathogen or tumor, optionally in combination with an adjuvant, effective to immunize the animal.

[0143] To prepare expression cassettes for transformation herein, the recombinant DNA sequence or segment may be circular or linear, double-stranded or single-stranded. A DNA sequence which encodes an RNA sequence that is substantially complementary to a mRNA sequence encoding a gene product of interest is typically a "sense" DNA sequence cloned into a cassette in the opposite orientation (i.e., 3

5 as plasmid DNA, that can also contain coding regions flanked by control sequences which promote the expression of the DNA in a cell. As used herein, "chimeric" means that a vector comprises DNA from at least two different species, or comprises DNA from the same species, which is linked or associated in a manner which does not occur in the "native" or wild-type of the species.

[0144] Aside from DNA sequences that serve as transcription units, or portions thereof, a portion of the DNA may be untranscribed, serving a regulatory or a structural function. For example, the DNA may itself comprise a promoter that is active in eukaryotic cells, e.g., mammalian cells, or in certain cell types, or may utilize a promoter already present in the genome that is the transformation target of the lymphotropic virus. Such promoters include the CMV promoter, as well as the SV40 late promoter and retroviral LTRs (long terminal repeat elements), e.g., the MMTV, RSV, MLV or HIV LTR, although many other promoter elements well known to the art may be employed in the practice of the invention.

[0145] Other elements functional in the host cells, such as introns, enhancers, polyadenylation sequences and the like, may also be a part of the recombinant DNA. Such elements may or may not be necessary for the function of the DNA, but may provide improved expression of the DNA by affecting transcription, stability of the mRNA, or the like. Such elements may be included in the DNA as desired to obtain the optimal performance of the transforming DNA in the cell.

[0146] The recombinant DNA to be introduced into the cells may contain either a selectable marker gene or a reporter gene or both to facilitate identification and selection of transformed cells from the population of cells sought to be transformed. Alternatively, the selectable marker may be carried on a separate piece of DNA and used in a co-transformation procedure. Both selectable markers and reporter genes may be flanked with appropriate regulatory sequences to enable expression in the host cells. Useful selectable markers are well known in the art and include, for example, antibiotic and herbicide-resistance genes, such as neo, hpt, dhfr, bar, aroA, puro, hyg, dapA and the like. See also, the genes listed on Table 1 of Lundquist et al. (U.S. Pat. No. 5,848,956).

[0147] Reporter genes are used for identifying potentially transformed cells and for evaluating the functionality of regulatory sequences. Reporter genes which encode for easily assayable proteins are well known in the art. In general, a reporter gene is a gene which is not present in or expressed by the recipient organism or tissue and which encodes a protein whose expression is manifested by some easily detectable property, e.g., enzymatic activity. Exemplary reporter genes include the chloramphenicol acetyl transferase gene (cat) from Tn9 of E. coli, the beta-glucuronidase gene (gus) of the uidA locus of E. coli, the green, red, or blue fluorescent protein gene, and the luciferase gene. Expression of the reporter gene is assayed at a suitable time after the DNA has been introduced into the recipient cells.

[0148] The general methods for constructing recombinant DNA which can transform target cells are well known to those skilled in the art, and the same compositions and methods of construction may be utilized to produce the DNA useful herein. For example, Sambrook et al., Molecular Cloning: A Laboratory Manual (2002) provides suitable methods of construction.

[0149] The recombinant DNA can be readily introduced into the host cells, e.g., mammalian, yeast or insect cells, by transfection with an expression vector comprising the recombinant DNA by any procedure useful for the introduction into a particular cell, e.g., physical or biological methods, to yield a transformed (transgenic) cell having the recombinant DNA so that the DNA sequence of interest is expressed by the host cell. In one embodiment, at least one of the recombinant DNA which is introduced to a cell is maintained extrachromosomally. In one embodiment, at least one recombinant DNA is stably integrated into the host cell genome.

[0150] Physical methods to introduce a recombinant DNA into a host cell include calcium-mediated methods, lipofection, particle bombardment, microinjection, electroporation, and the like. Biological methods to introduce the DNA of interest into a host cell include the use of DNA and RNA viral vectors. Viral vectors, e.g., retroviral or lentiviral vectors, have become a widely used method for inserting genes into eukaryotic, such as mammalian, e.g., human, cells. Other viral vectors useful to introduce genes into cells can be derived from poxviruses, e.g., vaccinia viruses, herpes viruses, adenoviruses, adeno-associated viruses, baculoviruses, and the like.

[0151] To confirm the presence of the recombinant DNA sequence in the host cell, a variety of assays may be performed. Such assays include, for example, molecular biological assays well known to those of skill in the art, such as Southern and Northern blotting, RT-PCR and PCR; biochemical assays, such as detecting the presence or absence of a particular gene product, e.g., by immunological means (ELISAs and Western blots) or by other molecular assays.

[0152] To detect and quantitate RNA produced from introduced recombinant DNA segments, RT-PCR may be employed. In this application of PCR, it is first necessary to reverse transcribe RNA into DNA, using enzymes such as reverse transcriptase, and then through the use of conventional PCR techniques amplify the DNA. In most instances PCR techniques, while useful, will not demonstrate integrity of the RNA product. Further information about the nature of the RNA product may be obtained by Northern blotting. This technique demonstrates the presence of an RNA species and gives information about the integrity of that RNA. The presence or absence of an RNA species can also be determined using dot or slot blot Northern hybridizations. These techniques are modifications of Northern blotting and only demonstrate the presence or absence of an RNA species.

[0153] While Southern blotting and PCR may be used to detect the recombinant DNA segment in question, they do not provide information as to whether the recombinant DNA segment is being expressed. Expression may be evaluated by specifically identifying the peptide products of the introduced DNA sequences or evaluating the phenotypic changes brought about by the expression of the introduced DNA segment in the host cell.

[0154] The recombinant viruses described herein have modifications in genomic sequences relative to a corresponding wild-type viral genome, i.e., the genome of the recombinant virus has a modification which includes a deletion, and optionally an insertion, in a region corresponding to sequences for a viral protein that is associated with transcription, is nonstructural or nonglycosylated, or is a glycoprotein. The mutation in the viral genome is effective to inhibit or prevent production of at least one functional viral protein from that genome when those sequences are present in a nontransgenic cell which supports viral replication. In one embodiment, the deletion includes from 1 up to thousands of nucleotides, e.g., 1%, 10%, 50%, 90% or more of sequences corresponding to the coding region for the viral protein. In one embodiment, the deleted sequences correspond to sequences with a substantial identity, e.g., at least 80% or more, e.g., 85%, 90% or 95% and up to 100% or any integer in between, nucleic acid sequence identity, to VP30 sequences.

[0155] In one embodiment, the viral genome in an infectious, replication-incompetent negative-sense, single-stranded RNA virus of the invention includes a deletion in sequences corresponding to those in a wild-type viral genome for a protein that is associated with transcription or is nonstructural or nonglycoslyated, or is a glycoprotein, and includes heterologous sequences that are nontoxic to host cells including cells in an organism to be immunized. In one embodiment, the heterologous sequence is a marker sequence, a selectable sequence or other sequence which is detectable or capable of detection, e.g., GFP or luciferase, or a selectable gene such as an antibiotic resistance gene, e.g., a hygromycin B resistance gene or neomycin phosphotransferase gene, which marker gene or selectable gene is not present in the host cell prior to introduction of the vector.

Pharmaceutical Compositions

[0156] Pharmaceutical anti-viral compositions of the present invention, suitable for administration, e.g., nasal, parenteral or oral administration, such as by intravenous, intramuscular, topical or subcutaneous routes, optionally further comprising sterile aqueous or non-aqueous solutions, suspensions, and emulsions. The compositions can further comprise auxiliary agents or excipients, as known in the art. The composition of the invention is generally presented in the form of individual doses (unit doses).

[0157] Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and/or emulsions, which may contain auxiliary agents or excipients known in the art. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Carriers or occlusive dressings can be used to increase skin permeability and enhance antigen absorption. Liquid dosage forms for oral administration may generally comprise a liposome solution containing the liquid dosage form. Suitable forms for suspending liposomes include emulsions, suspensions, solutions, syrups, and elixirs containing inert diluents commonly used in the art, such as purified water. Besides the inert diluents, such compositions can also include adjuvants, wetting agents, emulsifying and suspending agents, or sweetening, flavoring, or perfuming agents.

[0158] When a composition of the present invention is used for administration to an individual, it can further comprise salts, buffers, adjuvants, or other substances which are desirable for improving the efficacy of the composition. For vaccines, adjuvants, substances which can augment a specific immune response, can be used. Normally, the adjuvant and the composition are mixed prior to presentation to the immune system, or presented separately, but into the same site of the organism being immunized.

[0159] In one embodiment, the pharmaceutical composition is part of a controlled release system, e.g., one having a pump, or formed of polymeric materials (see Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Fla. (1974); Controlled Drug Bioavailability, Drug Product Design and Performance, Smolen and Ball (eds.), Wiley, New York (1984); Ranger & Peppas, J. Macromol. Sci. Rev. Macromol. Chem., 23:61 (1983); see also Levy et al., Science, 228:190 (1985); During et al., Ann. Neurol., 25:351 (1989); Howard et al., J. Neurosurg., 71:105 (1989)). Other controlled release systems are discussed in the review by Langer (Science, 249:1527 (1990)).

[0160] The pharmaceutical compositions of the present invention comprise a therapeutically effective amount of one or more anti-viral compounds, for instance, those identified by the screening methods of the invention, and a pharmaceutically acceptable carrier. In a specific embodiment, the term "pharmaceutically acceptable" means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeiae for use in animals, and more particularly in humans. The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the pharmaceutical composition is administered. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. These compositions can take the form of solutions, suspensions, emulsion, tablets, pills, capsules, powders, sustained-release formulations and the like. These compositions can be formulated as a suppository. Oral formulation can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, etc. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E. W. Martin. Such compositions will contain a therapeutically effective amount of the virus, such as one in purified form, together with a suitable amount of carrier so as to provide the form for proper administration to the patient. The formulation should suit the mode of administration.

[0161] The compositions may be systemically administered, e.g., orally, in combination with a pharmaceutically acceptable vehicle such as an inert diluent. For oral administration, the compound(s) may be combined with one or more excipients and used in the form of ingestible capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions should contain at least 0.1% of active compound. The percentage of the compositions and preparations may, of course, be varied and may conveniently be between about 2 to about 60% of the weight of a given unit dosage form. The amount of active compound in such useful compositions is such that an effective dosage level will be obtained.

[0162] The compositions may also contain the following: binders such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, fructose, lactose or aspartame or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring may be added. Various other materials may be present. For instance, a syrup or elixir may contain the virus, sucrose or fructose as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavoring such as cherry or orange flavor. Of course, any material used in preparing any unit dosage form, including sustained-release preparations or devices, should be pharmaceutically acceptable and substantially non-toxic in the amounts employed. The composition also be administered intravenously or intraperitoneally by infusion or injection. Solutions of the compound(s) can be prepared in water or a suitable buffer, optionally mixed with a nontoxic surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of undesirable microorganisms.

[0163] The pharmaceutical dosage forms suitable for injection or infusion can include sterile aqueous solutions or dispersions or sterile powders comprising the active ingredient which are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. In all cases, the ultimate dosage form should be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions or by the use of surfactants. The prevention of the action of undesirable microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it may be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride.

[0164] Sterile injectable solutions are prepared by incorporating the compound(s) in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filter sterilization.

[0165] Useful liquid carriers include water, alcohols or glycols or water-alcohol/glycol blends, in which the present compound(s) can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants. Adjuvants such as fragrances and additional antimicrobial agents can be added to optimize the properties for a given use. The resultant liquid compositions can be applied from absorbent pads, used to impregnate bandages and other dressings, or sprayed onto the affected area using pump-type or aerosol sprayers.

[0166] Useful dosages of the compositions of the invention can be determined by comparing their in vitro activity and in vivo activity in animal models.

Pharmaceutical Purposes

[0167] The administration of the composition may be for either a "prophylactic" or "therapeutic" purpose. When provided prophylactically, the compositions of the invention are provided before any symptom or clinical sign of a pathogen infection becomes manifest. The prophylactic administration of the composition serves to prevent or attenuate any subsequent infection.

[0168] When provided therapeutically, the compositions of the invention are provided upon the detection of a symptom or clinical sign of actual infection. The therapeutic administration of the compound(s) serves to attenuate any actual infection.

[0169] Thus, a composition of the present invention may be provided either before the onset of infection (so as to prevent or attenuate an anticipated infection) or after the initiation of an actual infection.

[0170] A composition is said to be "pharmacologically acceptable" if its administration can be tolerated by a recipient mammal. Such an agent is said to be administered in a "therapeutically effective amount" if the amount administered is physiologically significant. A composition of the present invention is physiologically significant if its presence results in a detectable change in the physiology of a recipient patient, e.g., enhances at least one primary or secondary humoral or cellular immune response against at least one strain of a virus.

[0171] The "protection" provided need not be absolute, i.e., the infection need not be totally prevented or eradicated, if there is a statistically significant improvement compared with a control population or set of mammals. Protection may be limited to mitigating the severity or rapidity of onset of symptoms or clinical signs of the virus infection.

Exemplary Compounds and Formulations

[0172] Compounds useful in methods of the invention include, but are not limited to, triphenylethylenes, tamoxifen and derivatives thereof such as raloxifene and clomiphene, calcium channel blockers, tetranortriterpenoids, antipsychotics, sigma receptor agonists, anticholinergics, steroids, inhibitor of calcium-independent phospholipase A.sub.2, inhibitors of magnesium-dependent phosphatidate phosphohydrolase, inhibitors of the inducible microsomal PGE.sub.2 synthase, inhibitors of Hsp90, and dopamine antagonists.

[0173] In one embodiment, the compound may be a compound of formula (I):

##STR00001##

wherein

[0174] X is O or NH;

[0175] each R.sup.1 is independently aryl, heteroaryl, (C.sub.1-C.sub.10)alkyl, or (C.sub.1-C.sub.10)alkaryl;

[0176] R.sup.2 is (C.sub.1-C.sub.10)alkyl; and

[0177] any aryl, heteroaryl, alkyl of R.sup.1 and R.sup.2 can optionally be substituted with one or more (e.g., one, two, three, four, five, etc.) hydroxy, halo, carboxy, nitro, amino, phenyl, or trifluoromethyl groups;

[0178] or a salt thereof.

[0179] The various salts of formula I, and of formulas II-VII below, can be formed from, for example, pharmaceutically acceptable acids such as methane sulfonic acid, benzene sulfonic acid, or toluene sulfonic acid. In one specific embodiment, the compound of formula (I) is benztropine mesylate:

##STR00002##

[0180] In another embodiment, the compound may be a compound of formula (II):

##STR00003##

wherein

[0181] each R.sup.1 is independently hydrogen, hydroxy, halo, carboxy, nitro, amino, trifluoromethyl, aryl, heteroaryl, (C.sub.1-C.sub.10)alkyl, or (C.sub.1-C.sub.10)alkaryl; and

[0182] any aryl, heteroaryl, or alkyl of R.sup.1 can optionally be substituted with one or more (e.g., one, two, three, four, five, etc.) hydroxy, halo, carboxy, nitro, amino, phenyl, or trifluoromethyl groups;

[0183] or a salt thereof.

[0184] In one specific embodiment, the compound of formula (II) may be Fluspirilene (8-[4,4-bis(4-fluorophenyl)butyl]-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-- one):

##STR00004##

[0185] In another embodiment, the compound may be a compound of formula (III):

##STR00005##

wherein

[0186] X is O or NH;

[0187] each Y is independently hydrogen or halo;

[0188] each R.sup.1 is independently hydrogen, hydroxy, halo, carboxy, nitro, amino, trifluoromethyl, aryl, heteroaryl, (C.sub.1-C.sub.10)alkyl, or (C.sub.1-C.sub.10)alkaryl; and

[0189] any aryl, heteroaryl, or alkyl of R.sup.1 can optionally be substituted with one or more (e.g., one, two, three, four, five, etc.) hydroxy, halo, carboxy, nitro, amino, phenyl, or trifluoromethyl groups;

[0190] or a salt thereof.

[0191] In one specific embodiment, the compound of formula (III) may be B1552 bromoenol lactone:

##STR00006##

[0192] In another embodiment, the compound may be a compound of formula (IV):

##STR00007##

wherein

[0193] each R.sup.1 is independently --X--R.sup.2;

[0194] each X is independently O, NH, or a direct bond;

[0195] each R.sup.2 is independently hydrogen, hydroxy, halo, carboxy, nitro, amino, trifluoromethyl, aryl, heteroaryl, (C.sub.1-C.sub.10)alkyl, or (C.sub.1-C.sub.10)alkaryl; and

[0196] any aryl, heteroaryl, or alkyl of R.sup.2 can optionally be substituted with one or more (e.g., one, two, three, four, five, etc.) hydroxy, halo, nitro, amino, phenyl, or trifluoromethyl groups;

[0197] or a salt thereof.

[0198] In one specific embodiment, the compound of formula (IV) may be cortexolone:

##STR00008##

[0199] In another embodiment, the compound may be a compound of formula (V):

##STR00009##

wherein

[0200] each R.sup.1 is independently --X--R.sup.2;

[0201] each X is independently O, NH, or a direct bond;

[0202] each R.sup.2 is independently hydrogen, hydroxy, halo, carboxy, nitro, amino, trifluoromethyl, aryl, heteroaryl, (C.sub.1-C.sub.10)alkyl, or (C.sub.1-C.sub.10)alkaryl; and

[0203] any aryl, heteroaryl, or alkyl of R.sup.2 can optionally be substituted with one or more (e.g., one, two, three, four, five, etc.) hydroxy, halo, carboxy, nitro, amino, phenyl, or trifluoromethyl groups;

[0204] or a salt thereof.

[0205] In one specific embodiment, the compound of formula (V) may be (R,R)-cis-diethyltetrahydro-2,8-chrysenediol:

##STR00010##

[0206] In another embodiment, the compound may be a compound of formula (VI):

##STR00011##

wherein

[0207] each R.sup.1 is independently --X--R.sup.2;

[0208] each X is independently O, NH, or a direct bond;

[0209] each R.sup.2 is independently hydrogen, hydroxy, halo, carboxy, nitro, amino, trifluoromethyl, carboxy, aryl, heteroaryl, (C.sub.1-C.sub.10)alkyl, or (C.sub.1-C.sub.10)alkaryl; and

[0210] any aryl, heteroaryl, or alkyl of R.sup.2 can optionally be substituted with one or more (e.g., one, two, three, four, five, etc.) hydroxy, halo, carboxy, nitro, amino, phenyl, or trifluoromethyl groups;

[0211] or a salt thereof.

[0212] In two specific embodiments, the compound of formula (VI) may be:

##STR00012##

In another embodiment, the compound may be a compound of formula (VII):

##STR00013##

wherein

[0213] each R.sup.1 is independently --X--R.sup.2;

[0214] each X is independently O, NH, or a direct bond;

[0215] each R.sup.2 is independently hydrogen, hydroxy, halo, carboxy, nitro, amino, trifluoromethyl, carboxy, aryl, heteroaryl, (C.sub.1-C.sub.10)alkyl, or (C.sub.1-C.sub.10)alkaryl; and

[0216] any aryl, heteroaryl, or alkyl of R.sup.2 can optionally be substituted with one or more (e.g., one, two, three, four, five, etc.) hydroxy, halo, carboxy, nitro, amino, phenyl, or trifluoromethyl groups;

[0217] or a salt thereof.

[0218] In one specific embodiment, the compound of formula (VII) may be L-687,384 (1-benzyl-spiro(1,2,3,4-tetrahydronaphthalene-1,4-piperidine):

##STR00014##

[0219] In another embodiment, the compound may be a compound of formula (VIII)

##STR00015##

wherein Z is C.dbd.O or a covalent bond; Y is H or O(C.sub.1-C.sub.4)alkyl, R.sup.1 and R.sup.2 are individually (C.sub.1-C.sub.4)alkyl or together with N are a saturated heterocyclic group, R.sup.3 is ethyl or chloroethyl, R.sup.4 is H, R.sup.5 is I, O(C.sub.1-C.sub.4)alkyl or H and R.sup.6 is I, O(C.sub.1-C.sub.4)alkyl or H with the proviso that when R.sup.4, R.sup.5, and R.sup.6 are H, R.sup.3 is not ethyl; or a pharmaceutically acceptable salt, including mixtures thereof.

[0220] In another embodiment, the compound can be a compound of formula (IX):

##STR00016##

wherein

[0221] Ar is a substituted or unsubstituted aryl or heteroaryl moiety;

[0222] X is --O--, --NH--, --NRx-, --CH.sub.2--, --CHRx-, or --C(Rx).sub.2-, wherein Rx is a hydrogen, a halogen, a protecting group, an aliphatic moiety, a heteroaliphatic moiety, an acyl moiety; an aryl moiety; a heteroaryl moiety; hydroxy, alkoxy; aryloxy; thioxy; alkylthio; arylthio; heteroaryloxy; or heteroarylthio moiety;

[0223] a dashed line represents either the presence or absence of a bond;

[0224] R.sub.1 is hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; --OH; --OR.sub.A; --C(.dbd.O)R.sub.A; --CHO; --CO.sub.2H; --CO.sub.2R.sub.A; --CN; --SCN; --SR.sub.A; --SOR.sub.A; --SO.sub.2R.sub.A; --NO.sub.2; --N.sub.3; --NH.sub.2; --NHR.sub.A; --N(R.sub.A).sub.2; --NHC(.dbd.O)R.sub.A; --NR.sub.AC(.dbd.O)R.sub.A; --NR.sub.AC(.dbd.O)N(R.sub.A).sub.2; --OC(.dbd.O)OR.sub.A; --OC(.dbd.O)R.sub.A; --OC(.dbd.O)N(R.sub.A).sub.2; --NR.sub.AC(.dbd.O)OR.sub.A; or --C(R.sub.A).sub.3; wherein each occurrence of R.sub.A is independently a hydrogen, a halogen, a protecting group, an aliphatic moiety, a heteroaliphatic moiety, an acyl moiety; an aryl moiety; a heteroaryl moiety; hydroxy, alkoxy; aryloxy; thioxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;

[0225] R.sub.2 is hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; --OH; --OR.sub.B; --C(.dbd.O)R.sub.B; --CHO; --CO.sub.2H; --CO.sub.2R.sub.B; --CN; --SCN; --SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B; --NO.sub.2; --N.sub.3; --NH.sub.2; --NHR.sub.B; --N(R.sub.B).sub.2; --NHC(.dbd.O)R.sub.B; --NR.sub.BC(.dbd.O)R.sub.B; --NR.sub.BC(.dbd.O)N(R.sub.B).sub.2; --OC(.dbd.O)N(R.sub.B).sub.2; --NR.sub.BC(.dbd.O)OR.sub.B; or --C(R.sub.B).sub.3; wherein each occurrence of R.sub.B is independently a hydrogen, a halogen, a protecting group, an aliphatic moiety, a heteroaliphatic moiety, an acyl moiety; an aryl moiety; a heteroaryl moiety; hydroxy, alkoxy; aryloxy; thioxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety; or R.sub.1 and R.sub.2 taken together form an epoxide ring, aziridine ring, cyclopropyl ring, or a bond of a carbon-carbon double bond;

[0226] R.sub.3 is hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; --OH; --OR.sub.C; --C(.dbd.O)R.sub.C; --CHO; --CO.sub.2H; --CO.sub.2R.sub.C; --CN; --SCN; --SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C; --NO.sub.2; --N.sub.3; --NH.sub.2; --NHR.sub.C; --N(R.sub.C).sub.2; --NHC(.dbd.O)R.sub.C; --NR.sub.CC(.dbd.O)R.sub.C; --NR.sub.CC(.dbd.O)N(R.sub.C).sub.2; --OC(.dbd.O)OR.sub.C; --OC(.dbd.O)R.sub.C; --OC(.dbd.O)N(R.sub.C).sub.2; --NR.sub.CC(.dbd.O)OR.sub.C; or --C(R.sub.C).sub.3; wherein each occurrence of R.sub.C is independently a hydrogen, a halogen, a protecting group, an aliphatic moiety, a heteroaliphatic moiety, an acyl moiety; an aryl moiety; a heteroaryl moiety; hydroxy, alkoxy; aryloxy; thioxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;

[0227] R.sub.4 is hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; --OH; --OR.sub.D; --C(.dbd.O)R.sub.D; --CHO; --CO.sub.2H; --CO.sub.2R.sub.D; --CN; --SCN; --SR.sub.D; --SOR.sub.D; --SO.sub.2R.sub.D; --NO.sub.2; --N.sub.3; --NH.sub.2; --NHR.sub.D; --N(R.sub.D).sub.2; --NHC(.dbd.O)R.sub.D; --NR.sub.DC(.dbd.O)R.sub.D; NR.sub.DC(.dbd.O)N(R.sub.D).sub.2; --OC(.dbd.O)OR.sub.D; --OC(.dbd.O)R.sub.D; --OC(.dbd.O)N(R.sub.D).sub.2; --OC(.dbd.O)OR.sub.D; --OC(.dbd.O)R.sub.D; --OC(.dbd.O)N(R.sub.D).sub.2; --NR.sub.DC(.dbd.O)OR.sub.D; or --C(R.sub.D).sub.3; wherein each occurrence of R.sub.D is independently a hydrogen, a halogen, a protecting group, an aliphatic moiety, a hetero aliphatic moiety, an acyl moiety; an aryl moiety; a heteroaryl moiety; hydroxy, alkoxy; aryloxy; thioxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;

[0228] R.sub.5 is hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; --OH; --OR.sub.E; --C(.dbd.O)R.sub.E; --CHO; --CO.sub.2H; --CO.sub.2R.sub.E; --CN; --SCN; --SR.sub.E; --SOR.sub.E; --SO.sub.2R.sub.E; --NO.sub.2; --N.sub.3; --NH.sub.2; --NHR.sub.E; --N(R.sub.E).sub.2; --NHC(.dbd.O)R.sub.E; --NR.sub.EC(.dbd.O)R.sub.E; --NR.sub.EC(.dbd.O)N(R.sub.E).sub.2; --OC(.dbd.O)OR.sub.E; --OC(.dbd.O)R.sub.E; --OC(.dbd.O)N(R.sub.E).sub.2; --NR.sub.EC(.dbd.O)OR.sub.E; or --C(R.sub.E).sub.3; wherein each occurrence of R.sub.E is independently a hydrogen, a halogen, a protecting group, an aliphatic moiety, a heteroaliphatic moiety, an acyl moiety; an aryl moiety; a heteroaryl moiety; hydroxy, alkoxy; aryloxy; thioxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;

[0229] R.sub.6 is hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; --OH; --OR.sub.F; --C(.dbd.O)R.sub.F; --CHO; --CO.sub.2H; --CO.sub.2R.sub.F; --CN; --SCN; --SR.sub.F; --SOR.sub.F; --SO.sub.2R.sub.F; --NO.sub.2; --N.sub.3; --NH.sub.2; --NHR.sub.F; --N(R.sub.F).sub.2; --NHC(.dbd.O)R.sub.F; --NR.sub.FC(.dbd.O)R.sub.F; --NR.sub.FC(.dbd.O)N(R.sub.F).sub.2; --OC(.dbd.O)OR.sub.F; --OC(.dbd.O)R.sub.F; --OC(.dbd.O)R.sub.F; --OC(.dbd.O)N((R.sub.F).sub.2; --NR.sub.FC(.dbd.O)OR.sub.F; or --C(R.sub.F).sub.3; wherein each occurrence of R.sub.F is independently a hydrogen, a halogen, a protecting group, an aliphatic moiety, a heteroaliphatic moiety, an acyl moiety; an aryl moiety; a heteroaryl moiety; hydroxy, alkoxy; aryloxy; thioxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;

[0230] R.sub.7 is hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; --OH; --OR.sub.G; --C(.dbd.O)R.sub.G; --CHO; --CO.sub.2H; --CO.sub.2R.sub.G; --CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G; --NO.sub.2; --N.sub.3; --NH.sub.2; --NHR.sub.G; --N(R.sub.G).sub.2; --NHC(.dbd.O)R.sub.G; --NR.sub.GC(.dbd.O)R.sub.G; --NR.sub.GC(.dbd.O)N(R.sub.G).sub.2; --OC(.dbd.O)OR.sub.G; --OC(.dbd.O)R.sub.G; --OC(.dbd.O)N(R.sub.G).sub.2; --NR.sub.GC(.dbd.O)OR.sub.G; or --C(R.sub.G).sub.3; wherein each occurrence of R.sub.G is independently a hydrogen, a halogen, a protecting group, an aliphatic moiety, a heteroaliphatic moiety, an acyl moiety; an aryl moiety; a heteroaryl moiety; hydroxy, alkoxy; aryloxy; thioxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;

[0231] R.sub.8 is hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or [mu]nsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; --OH; --OR.sub.H; --C(.dbd.O)R.sub.H; --CHO; --CO.sub.2H; --CO.sub.2R.sub.H; --CN; --SCN; --SR.sub.H; --SOR.sub.H; --SO.sub.2R.sub.H; --NO.sub.2; --N.sub.3; --NH.sub.2; --NHR.sub.H; --N(R.sub.H).sub.2; --NHC(.dbd.O)R.sub.H; --NR.sub.HC(.dbd.O)R.sub.H; --NR.sub.H(C.dbd.O)N(2R.sub.H).sub.2; --OC(.dbd.O)OR.sub.H; --O(C.dbd.O)R.sub.H; --NR.sub.HC(.dbd.O)OR.sub.H; or --C(R.sub.H).sub.3; wherein each occurrence of R.sub.H is independently a hydrogen, a halogen, a protecting group, an aliphatic moiety, a heteroaliphatic moiety, an acyl moiety; an aryl moiety; a heteroaryl moiety; hydroxy, alkoxy; aryloxy; thioxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;

[0232] R.sub.9 is hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; --OH; --OR.sub.I; .dbd.O; --C(.dbd.O)R.sub.I; --CHO; --CO.sub.2H; --CO.sub.2R.sub.I; --CN; --SCN; --SR.sub.I; --SOR.sub.I; --SO.sub.2R.sub.I; --NO.sub.2; --N.sub.3; --NH.sub.2; --NHR.sub.I; --N(R.sub.I).sub.2; --NHC(.dbd.O)R.sub.I; --NR.sub.IC(.dbd.O)R.sub.I; --NR.sub.IC(.dbd.O)N(R.sub.I).sub.2; --OC(.dbd.O)OR.sub.I; --OC(.dbd.O)R.sub.I; --OC(.dbd.O)N(R.sub.I).sub.2; --NR.sub.IC(.dbd.O)OR.sub.I; or --C(R.sub.I).sub.3; wherein each occurrence of R.sub.I is independently a hydrogen, a halogen, a protecting group, an aliphatic moiety, a heteroaliphatic moiety, an acyl moiety; an aryl moiety; a heteroaryl moiety; hydroxy, alkoxy; aryloxy; thioxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;

[0233] R.sub.10 is hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; --OH; --OR.sub.J; .dbd.O; --C(.dbd.O)R.sub.J; --CHO; --CO.sub.2H; --CO.sub.2R.sub.J; --CN; --SCN; --SR.sub.J; --SOR.sub.J; --SO.sub.2R.sub.J; --NO.sub.2; --N.sub.3; --NH.sub.2; --NHR.sub.J; --N(R.sub.J).sub.2; --NHC(.dbd.O)R.sub.J; --NR.sub.JC(.dbd.O)R.sub.J; --NR.sub.JC(.dbd.O)N(R.sub.J).sub.2; --OC(.dbd.O)OR.sub.J; --OC(.dbd.O)R.sub.J; --OC(.dbd.O)N(R.sub.J).sub.2; --NR.sub.JC(.dbd.O)OR.sub.J; or --C(R.sub.J).sub.3; wherein each occurrence of R.sub.J is independently a hydrogen, a halogen, a protecting group, an aliphatic moiety, a heteroaliphatic moiety, an acyl moiety; an aryl moiety; a heteroaryl moiety; hydroxy, alkoxy; aryloxy; thioxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or hetero arylthio moiety;

[0234] R.sub.11 is hydrogen, halo, hydroxy, or a carbonyl;

or a pharmaceutically acceptable salt, stereoisomer, tautomer, or pro-drug thereof.

[0235] In yet another embodiment, the compound (e.g., the compound of formula IX) can be a compound of formula (X):

##STR00017##

wherein

[0236] R.sub.6 is hydrogen; hydroxy; oxo (.dbd.O); or acetyl-protected hydroxyl; and

[0237] R.sub.9 is hydrogen; hydroxy; oxo (.dbd.O); or acetyl-protected hydroxyl.

[0238] In various embodiments, compounds of formulas (IX) and (X) can include gedunin, gedunol, epoxygedunin, 1,2.alpha.-epoxy-7-deacetoxy-7-oxodihydrogedunin, dihydrogedunin, 3.beta.-acetoxydeoxodihydrogedunin, 3.alpha.-hydroxydeoxodihydrogedunin, deacetoxy-7-oxogedunin, 3.beta.-hydroxydeoxodihydrogedunin, deacetoxy-7-oxogedunin, 1,2.alpha.-epoxydeacetoxydihydrogedunin, 3.beta.-hydroxydeoxydesacetoxy-7-oxogedunin, 7-deacetoxy-3-deacetyl-7-oxokhivorin, 1,3-dideacetyl-7-deacetoxy-7-oxokhivorin, tridesacetoxykhivorin, 1,3-dideacetylkhivorin, and/or Heudelottin C.

[0239] In another embodiment, the compound can be a compound of formula (XI):

##STR00018##

wherein

[0240] R.sup.1 is --X--R.sup.X;

[0241] X is O, NH, or a direct bond;

[0242] R.sup.x is hydrogen, hydroxy, halo, carboxy, nitro, amino, trifluoromethyl, aryl, heteroaryl, (C.sub.1-C.sub.10)alkyl, (C.sub.1-C.sub.10)alkenyl, or (C.sub.1-C.sub.10)alkylaryl;

[0243] R.sup.2 is hydrogen, trifluoromethyl, aryl, heteroaryl, (C.sub.1-C.sub.10)alkyl, (C.sub.1-C.sub.10)alkenyl, (C.sub.1-C.sub.10)alkylaryl, or an oxygen protecting group;

[0244] R.sup.3 is hydrogen, trifluoromethyl, aryl, heteroaryl, (C.sub.1-C.sub.10)alkyl, (C.sub.1-C.sub.10)alkenyl, (C.sub.1-C.sub.10)alkylaryl, or an oxygen protecting group;

[0245] R.sup.4 is hydrogen, trifluoromethyl, aryl, heteroaryl, (C.sub.1-C.sub.10)alkyl, (C.sub.1-C.sub.10)alkenyl, (C.sub.1-C.sub.10)alkylaryl, or an oxygen protecting group; and

[0246] any aryl, heteroaryl, or alkyl (e.g., of Rx, R.sup.2, R.sup.3, or R.sup.4) can optionally be substituted with one or more (e.g., one, two, three, four, five, etc.) hydroxy, halo, carboxy, nitro, amino, (C.sub.1-C.sub.10)alkylamino, di(C.sub.1-C.sub.10)alkylamino, (C.sub.1-C.sub.10)alkyl, (C.sub.1-C.sub.10)alkenyl, (C.sub.1-C.sub.10)alkoxy, phenyl, benzyl, or trifluoromethyl groups;

[0247] or a salt thereof.

[0248] In some embodiments, the compound of formula (XI) may be geldanamycin, 17-AAG, or 17-DMAG.

[0249] In another embodiment, the compound can be a compound of formula (XII):

##STR00019##

wherein

[0250] R.sup.1 is H, (C.sub.1-C.sub.10)alkyl or (C.sub.1-C.sub.10)alkylaryl;

[0251] R.sup.2 is H, (C.sub.1-C.sub.10)alkyl or (C.sub.1-C.sub.10)alkylaryl;

[0252] R.sup.3 is hydrogen, trifluoromethyl, aryl, heteroaryl, (C.sub.1-C.sub.10)alkyl, (C.sub.1-C.sub.10)alkylaryl, or an oxygen protecting group;

[0253] R.sup.4 is hydrogen, trifluoromethyl, aryl, heteroaryl, (C.sub.1-C.sub.10)alkyl, (C.sub.1-C.sub.10)alkylaryl, or an oxygen protecting group;

[0254] R.sup.5 is H, (C.sub.1-C.sub.10)alkyl or (C.sub.1-C.sub.10)alkylaryl;

[0255] R.sup.6 is H or --X--R.sup.X; X is O, NH, or a direct bond;

[0256] R.sup.7 is --X--R.sup.X; X is O, NH, or a direct bond;

[0257] each R.sup.x is independently hydrogen, hydroxy, halo, carboxy, nitro, amino, trifluoromethyl, aryl, heteroaryl, (C.sub.1-C.sub.10)alkyl, or (C.sub.1-C.sub.10)alkaryl; and

[0258] any aryl, heteroaryl, or alkyl can optionally be substituted with one or more (e.g., one, two, three, four, five, etc.) hydroxy, halo, carboxy, nitro, amino, phenyl, or trifluoromethyl groups;

[0259] or a salt thereof.

[0260] In one specific embodiment, the compound of formula (XII) may be CCT-018159 (4-[4-(2,3-dihydro-1,4-benzodioxin-6-yl)-5-methyl-1H-pyrazol-3-yl]-6-ethy- l-1,3-benzenediol):

##STR00020##

[0261] In another embodiment, the compound can be a compound of formula (XIII):

##STR00021##

wherein

[0262] R.sup.1 is H, (C.sub.1-C.sub.10)alkyl, aryl, or (C.sub.1-C.sub.10)alkylaryl;

[0263] R.sup.2 is H, (C.sub.1-C.sub.10)alkyl aryl, or (C.sub.1-C.sub.10)alkylaryl;

[0264] each R.sup.3 is independently H or --X--R.sup.X;

[0265] X is O, NH, or a direct bond;

[0266] each R.sup.x is independently hydrogen, hydroxy, halo, carboxy, nitro, amino, trifluoromethyl, aryl, heteroaryl, (C.sub.1-C.sub.10)alkyl, or (C.sub.1-C.sub.10)alkylaryl;

[0267] any aryl, heteroaryl, or alkyl can optionally be substituted with one or more (e.g., one, two, three, four, five, etc.) hydroxy, halo, carboxy, nitro, amino, phenyl, or trifluoromethyl groups; and

[0268] n is 1, 2, 3, 4, or 5;

[0269] or a salt thereof.

[0270] In one specific embodiment, the compound of formula (XIII) may be AEG 3482 (6-phenylimidazo[2,1-b]-1,3,4-thiadiazole-2-sulfonamide):

##STR00022##

[0271] The compounds of the invention, such as those having formulas (I)-(XIII), can be formulated as pharmaceutical compositions and administered to a mammalian host, such as a human patient in a variety of forms adapted to the chosen route of administration, i.e., orally or parenterally, by intravenous, intramuscular, topical or subcutaneous routes.

[0272] The present compounds may be systemically administered, e.g., orally, in combination with a pharmaceutically acceptable vehicle such as an inert diluent or an assimilable edible carrier. They may be enclosed in hard or soft shell gelatin capsules, may be compressed into tablets, or may be incorporated directly with the food of the patient's diet. For oral administration, the active compound may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and preparations should contain at least 0.1% of active compound. The percentage of the compositions and preparations may, of course, be varied and may conveniently be between about 2 to about 60% of the weight of a given unit dosage form. The amount of active compound in such useful compositions is such that an effective dosage level will be obtained.

[0273] The tablets, troches, pills, capsules, and the like may also contain the following: binders such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, fructose, lactose or aspartame or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring may be added. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier, such as a vegetable oil or a polyethylene glycol. Various other materials may be present as coatings or to otherwise modify the physical form of the solid unit dosage form. For instance, tablets, pills, or capsules may be coated with gelatin, wax, shellac or sugar and the like. A syrup or elixir may contain the active compound, sucrose or fructose as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavoring such as cherry or orange flavor. Of course, any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed. In addition, the active compound may be incorporated into sustained-release preparations and devices.

[0274] The active compound may also be administered intravenously or intraperitoneally by infusion or injection. Solutions of the active compound or its salts can be prepared in water, optionally mixed with a nontoxic surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

[0275] The pharmaceutical dosage forms suitable for injection or infusion can include sterile aqueous solutions or dispersions or sterile powders comprising the active ingredient which are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. In all cases, the ultimate dosage form should be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions or by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it may be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0276] Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filter sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the previously sterile-filtered solutions.

[0277] For topical administration, the present compounds may be applied in pure form, i.e., when they are liquids. However, it will generally be desirable to administer them to the skin as compositions or formulations, in combination with a dermatologically acceptable carrier, which may be a solid or a liquid.

[0278] Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silica, alumina and the like. Useful liquid carriers include water, alcohols or glycols or water-alcohol/glycol blends, in which the present compounds can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants. Adjuvants such as fragrances and additional antimicrobial agents can be added to optimize the properties for a given use. The resultant liquid compositions can be applied from absorbent pads, used to impregnate bandages and other dressings, or sprayed onto the affected area using pump-type or aerosol sprayers.

[0279] Thickeners such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses or modified mineral materials can also be employed with liquid carriers to form spreadable pastes, gels, ointments, soaps, and the like, for application directly to the skin of the user.

[0280] Useful dosages of the compounds of the invention can be determined by comparing their in vitro activity and in vivo activity in animal models. Methods for the extrapolation of effective dosages in mice, and other animals, to humans are known to the art; for example, see U.S. Pat. No. 4,938,949.

[0281] Generally, the concentration of the compounds of the invention in a liquid composition, such as a lotion, will be from about 0.1-25 wt-%, e.g., from about 0.5-10 wt-%. The concentration in a semi-solid or solid composition such as a gel or a powder will be about 0.1-5 wt-%, e.g., about 0.5-2.5 wt-%.

[0282] The amount of the compound, or an active salt or derivative thereof, required for use alone or with other compounds will vary not only with the particular salt selected but also with the route of administration, the nature of the condition being treated and the age and condition of the patient and will be ultimately at the discretion of the attendant physician or clinician.

[0283] In general, however, a suitable dose may be in the range of from about 0.5 to about 100 mg/kg, e.g., from about 10 to about 75 mg/kg of body weight per day, such as 3 to about 50 mg per kilogram body weight of the recipient per day, such as in the range of 6 to 90 mg/kg/day, or in the range of 15 to 60 mg/kg/day.

[0284] The compound may be conveniently administered in unit dosage form; for example, containing 5 to 1000 mg, conveniently 10 to 750 mg, most conveniently, 50 to 500 mg of active ingredient per unit dosage form.

[0285] The active ingredient may be administered to achieve peak plasma concentrations of the active compound of from about 0.5 to about 75 .mu.M, e.g., about 1 to 50 .mu.M, such as about 2 to about 30 .mu.M. This may be achieved, for example, by the intravenous injection of a 0.05 to 5% solution of the active ingredient, optionally in saline, or orally administered as a bolus containing about 1-100 mg of the active ingredient. Desirable blood levels may be maintained by continuous infusion to provide about 0.01-5.0 mg/kg/hr or by intermittent infusions containing about 0.4-15 mg/kg of the active ingredient(s).

[0286] The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations; such as multiple inhalations from an insufflator or by application of a plurality of drops into the eye.

[0287] The invention will be further described in the following nonlimiting examples.

Example 1

Methods and Materials

[0288] Cells and Cell Lines.

[0289] Vero cells (green monkey kidney cells) were grown in Eagle's minimal essential medium (MEM) supplemented with 10% fetal calf serum (FCS), L-glutamine, vitamins, nonessential amino acid solution and antibiotics. The VeroVP30 cell line was established by cotransfecting Vero cells with pCAG-VP30 (for the expression of VP30) and pPur, a protein expression plasmid for the puromycin resistance gene (Clontech, Mountain View, Calif.), using the transfection reagent TransIT LT-1 (Mirus, Madison, Wis.). Two days after transfection, puromycin-resistant cells were selected with 5 .mu.g/mL puromycin (Sigma, St. Louis, Mo.). Individual cell clones were screened for VP30 expression by flow cytometry with a polyclonal peptide antibody to VP30.

[0290] Human embryonic kidney 293T cells were grown in high-glucose Dulbecco's modified Eagle medium containing 10% FCS, L-glutamine, and antibiotics. All cells were maintained at 37.degree. C. and 5% CO.sub.2.

[0291] Flow Cytometry.

[0292] Cells were detached in phosphate-buffered saline (PBS) containing 0.02% EDTA and then washed once with cold PBS supplemented with 2% FCS and 0.1% sodium azide (wash buffer). Cells were incubated with a VP30 antibody on ice for 20 minutes. After washing in buffer, the cells were further incubated with a secondary antibody labeled with fluorescent isothiocyanate (Zymed Laboratories, Carlsbad, Calif.). They were then washed with buffer and analyzed by FACSCalibur with Cell Quest software (Becton Dickinson, Franklin Lakes, N.J.).

[0293] Generation of Ebola.DELTA.VP30 Viruses.

[0294] The plasmid pTM-T7G-Ebo-Rib, containing the full-length Ebolavirus cDNA flanked by T7 RNA polymerase promoter and ribozyme sequences, is described in Newmann et al. (2002). First, a fragment encompassing nucleotides 6180 to 10942 (numbers refers to the positive-sense antigenome) was subcloned into a kanamycin-resistant cloning vector. Next, the VP30 ORF was replaced with those encoding neo or eGFP, respectively, by a series of overlapping PCR amplification steps using Pfu Turbo (Stratagene, La Jolla, Calif.). The altered subgenomic fragments were transferred back into the full-length Ebolavirus cDNA plasmid using two unique restriction sites, SalI and SacI (FIGS. 1A-1B). The resultant plasmids, designated pTM-Ebola.DELTA.VP30-neo or -eGFP, were sequenced to verify the replacement of the VP30 ORF and the lack of any unwanted mutations.

[0295] To artificially generate Ebolavirus, 5.times.10.sup.5 293T cells were transfected with 1.0 .mu.g pTM-Ebola.DELTA.VP30, 2.0 .mu.g pCAG-L, 1.0 .mu.g pCAG-NP, 0.5 .mu.g pCAG-VP35, 0.5 .mu.g pCAG-VP30, and 1.0 .mu.g pCAG-T7 pol, using TransIT LT1 (Mirus, Madison, Wis.) in BSL-4 containment (Neumann et al., 2002). Five days after transfection, the supernatant was harvested, cellular debris removed by low speed centrifugation, and the virus amplified in VeroVP30 cells at 37.degree. C. and 5% CO.sub.2 with propagation medium containing 2% FCS in MEM supplemented with L-glutamine, vitamins, nonessential amino acid solution and antibiotics without puromycin.

[0296] Plaque Assay and Immunostaining Assay.

[0297] To determine the titers of wild-type Ebolavirus or Ebola.DELTA.VP30 viruses, tenfold dilutions of the viruses were absorbed to confluent VeroVP30 or wild-type Vero cells for 1 hour at 37.degree. C., after which any unbound virus was removed by washing cells with propagation medium. The cells were then overlaid with propagation medium containing 1.5% methyl cellulose (Sigma). Seven days after infection, cells were fixed with 10% buffered formaldehyde, taken out of BSL-4, permeabilized with 0.25% Triton X-100 in PBS for 10 minutes, and blocked with 4% goat serum and 1% bovine serum albumin (BSA) in PBS for 60 minutes. Cells were then incubated for 60 minutes with a 1:1000 dilution of a mouse anti-VP40 monoclonal antibody, washed with PBS, and incubated for 60 minutes with a 1:1000 dilution of an antimouse IgG-peroxidase-conjugated secondary antibody (Kirkegaard & Perry Laboratories Inc., Gaithersburg, Md.). After washing with PBS, cells were incubated with 3,3'-diaminobenzidine tetrahydrochloride (DAB, Sigma) in PBS. The reaction was stopped by rinsing cells with water.

[0298] Western Blotting.

[0299] Partially purified virus resuspended in lysis buffer (50 mM Tris-HCl [pH 7.5], 150 mM NaCl, 0.5% Triton X-100, and 0.1% SDS) containing protease inhibitors (complete protease inhibitor cocktails [Roche]) was incubated at 100.degree. C. for 5 minutes, taken out of BSL-4, and separated on 4-20% polyacrylamide gels. Resolved proteins were transferred to Western polyvinylidine difluoride membranes (Schleicher & Schuell, Sanford, Me.) and blocked overnight at 4.degree. C. with 5% skim milk in PBST (0.05% Tween 20 [Sigma] in PBS). Blots were incubated with primary antibodies (a mouse anti-NP antibody, a rabbit anti-VP35 antibody, a rabbit anti-VP40 antibody, a mouse anti-GP antibody, a rabbit anti-VP30 antibody, or a mouse anti-VP24 antibody) for 60 minutes at room temperature, washed three times with PBST, incubated with the appropriate secondary antibody conjugated to horseradish peroxidase (Zymed) for 60 minutes, and finally washed three times with PBST. Blots were then incubated in Lumi-Light Western blotting substrate (Roche, Indianapolis, Ind.) and exposed to X-rayfilm (Kodak, Rochester, N.Y.).

[0300] RNA Isolation and RT-PCR.

[0301] Cell culture supernatant from virus-infected VeroVP30 cells was inactivated with guanidinium isothiocyanate buffer and taken out of BSL-4. Viral RNA was isolated with the RNeasy Mini kit (Qiagen, Valencia, Calif.). RT-PCR was carried out with the RobusT One-Step RT-PCR kit (Finnzyme, Espoo, Finland), using 1 .mu.g of isolated RNA and Ebolavirus-specific primers. The resultant PCR products were cloned into pT7Blue (Novagen, San Diego, Calif.) and sequenced.

[0302] Transmission Electron Microscopy.

[0303] Ultrathin-section electronmicroscopy was performed as described in Noda et al. (2002). Briefly, at 36 hours postinfection, VeroVP30 cells infected with Ebola.DELTA.VP30-neo virus were fixed and inactivated with 2.5% glutaraldehyde in 0.1 M cacodylate buffer, taken out of BSL-4 and postfixed with 2% osmium tetroxide in the same buffer. Cells were then dehydrated with a series of ethanol gradients followed by propylene oxide, before being embedded in Epon 812 Resin mixture (TAAB Laboratories Equipment Ltd., Berkshire, UK). Thin sections were stained with 2% uranyl acetate and Raynold's lead, and examined under a HITACHI H-7500 electron microscope at 80 kV.

[0304] Selection of Escape Mutants.

[0305] Ebola.DELTA.VP30-eGFP was diluted tenfold (10.sup.-1 to 10.sup.-6) and incubated with the indicated mAbs at a concentration of 250 to 500 .mu.g of mAb/mL at 37.degree. C. for 60 minutes. The virus/mAb mixtures were inoculated onto VeroVP30 cells for 60 minutes. Viruses were amplified for 5 days in the presence of antibodies. Then, viruses that grew in the presence of mAbs (as determined by GFP expression) were harvested at the highest virus-positive dilution and passaged for a total of 3-6 times in the presence of antibodies. Viral RNA was isolated, RT-PCR amplified, and the GP sequence determined by sequence analysis.

Results

[0306] Generation and Passage of Ebola.DELTA.VP30-Neo Virus.

[0307] Previously a full-length cDNA clone of the Zaire ebolavirus-Mayinga was generated (Newmann et al., 2002). Using a subgenomic fragment that encompasses nucleotides 6180 to 10942 of the viral genome (numbers refers to the positive-sense antigenome), the ORF for VP30 was replaced with that of neomycin (neo), using a series of overlapping PCR amplification steps. After confirmation of the authenticity of the PCR fragments by sequence analysis, the altered subgenomic fragment was inserted into the full-length Ebolavirus cDNA construct via unique SalI and SacI restriction sites (FIGS. 1A-1B), resulting in an Ebolavirus cDNA genome deficient in the VP30 ORF. The artificial generation of Ebolavirus from plasmids is afforded by flanking this viral cDNA with T7 RNA polymerase promoter and hepatitis delta virus ribozyme sequences (Neumann et al., 2002).

[0308] To amplify VP30-deficient Ebolaviruses, a stable Vero E6 cell line (designated VeroVP30) was established by cotransfecting Vero cells with two protein expression plasmids encoding VP30 (pCAG-VP30) and puromycin (pPur, Clontech), and selecting cell clones resistant to 5.0 .mu.g/mL of puromycin. VP30 expression in individual clones was determined by flow cytometry with antibodies to VP30. The clone with the highest percentage of VP30-expressing cells (>90% as measured by flow cytometry, data not shown) was used in further studies to amplify Ebola.DELTA.VP30 viruses.

[0309] Ebola.DELTA.VP30-neo virus was rescued under BSL-4 conditions as described for wild-type Ebolavirus (Neumann et al., 2002). All work involving infectious Ebo.DELTA.VP30 viruses and all steps prior to inactivation of biological material were performed under BSL-4 conditions at the National Microbiology Laboratory of the Public Health Agency of Canada.

[0310] Briefly, human embryonic kidney (293T) cells were transfected with a plasmid for the transcription of the VP30-deficient Ebolavirus RNA, with plasmids for the expression of the Ebolavirus NP, VP30, VP35, and L proteins, and with a plasmid for the expression of T7 RNA polymerase. Five days after transfection, VeroVP30 cells were incubated with undiluted supernatant derived from plasmid-transfected cells. Seven days later, the supernatant was harvested, diluted tenfold, and used to infect fresh VeroVP30 cells for the next passage. A total of seven passages were carried out, using the highest dilution of the inoculum that still produced replicating viruses for each passage. The presence of replicating virus was assessed by cytopathic effects (CPE) and immunostaining of infected VeroVP30 cells with an antibody to VP40 (FIG. 2A, left panel). As a control, we also incubated the supernatants from each passage with wild-type Vero cells. As expected, CPE and viral antigens were undetectable in wild-type Vero cells (FIG. 2A, right panel), demonstrating that replicating Ebola.DELTA.VP30-neo virus was confined to VeroVP30 cells.

[0311] Although the manifestation of a CPE in infected VeroVP30 cells suggested the formation of infectious (but biologically contained) Ebolaviruses, further evidence was sought for the presence of virions in cell culture supernatant derived from infected VeroVP30 cells. Briefly, 5 days after VeroVP30 cells were infected with Ebola.DELTA.VP30-neo virus, supernatant was collected and partially purified over 20% sucrose. The pellet was suspended in PBS and separated on a 4-20% polyacrylamide gel. Western blot analyses were carried out with antibodies specific to the respective Ebolavirus protein. All viral proteins (with the exception of L, for which no antibody was available) were detected (FIG. 2B, `+` lanes). Note that VP30 protein in virions originates from VeroVP30 cells while the remaining proteins are encoded by Ebola.DELTA.VP30-neo virus. By contrast, no viral proteins were detected in a control sample derived from wild-type Vero cells infected with Ebola.DELTA.VP30-neo virus (FIG. 2B, `-` lanes).

[0312] Genetic Stability of Ebola.DELTA.VP30-Neo Virus.

[0313] A major concern with the use of VP30-deficient Ebolaviruses is the potential recombination with VP30 sequences integrated into the genome of the VeroVP30 helper cell line. Thus, to assess the genomic stability of Ebola.DELTA.VP30-neo virus, three independent passage experiments were performed (seven passages each). While Ebola.DELTA.VP30-neo virus replicated in VeroVP30 cells, viral replication was not observed in wild-type Vero cells. Total viral RNA was isolated from the cell culture supernatant of infected VeroVP30 cells after the seventh passage. A viral genomic fragment spanning the neo gene was amplified by RT-PCR, cloned and sequenced. A total of 20 clones were sequenced, and the sequences were identical to that of the Ebola.DELTA.VP30 cDNA construct used for virus generation. Hence, there was no evidence of recombination in any of three independent passage experiments, attesting to the genomic stability of the Ebola.DELTA.VP30-neo viral genome.

[0314] To further demonstrate the biosafety of Ebola.DELTA.VP30-neo virus, Ebola.DELTA.VP30-neo virus was collected after seven consecutive passages in VeroVP30 cells and this virus used for three consecutive "blind" passages in wild-type Vero cells. Briefly, Vero cells were infected at a multiplicity of infection (m.o.i.) of 5 with Ebola.DELTA.VP30-neo virus (passage 7). Six days later, supernatant was used for the next "blind" passage as well as for Western blot analysis. No viral NP protein was detected after any of the "blind" passages (data not shown). After three consecutive "blind" passages, plaque assays and immunostaining were carried out in wild-type Vero cells to confirm the absence of replicating Ebolavirus. As expected, replicating virus was not detected (data not shown). Collectively, these data further attest to the biosafety of the Ebola.DELTA.VP30 system.

[0315] Growth Kinetics of Ebola.DELTA.VP30-Neo Virus.

[0316] One of the major concerns raised by providing viral proteins in trans is that their amounts, expression kinetics or both may not match those found in cells infected with wild-type virus, leading to reduced virus titers and/or aberrant virion morphology. To address this potential pitfall, the growth kinetics of Ebola.DELTA.VP30-neo virus (FIG. 3, solid squares) were compared with that of wild-type Ebolavirus (FIG. 3, open circles). VeroVP30 cells (FIG. 3, top panels) or wild-type Vero cells (FIG. 3, bottom panels) were infected at a high m.o.i. of 1.0 or a low m.o.i. of 0.01 and supernatant was harvested every 24 hours. Virus titers of Ebola.DELTA.VP30-neo were determined in VeroVP30 cells, while virus titers of wild-type Ebolavirus were determined in wild-type Vero cells. To determine virus titers, cells were overlaid with 1.5% methylcellulose and 7 days later, assayed for VP40 expression using an immunostaining assay. Ebola.DELTA.VP30-neo virus replicated efficiently in VeroVP30 cells at both conditions tested, reaching 10.sup.7 focal-forming units (FFU)/ml on day 6 postinfection (FIG. 3, top panels, solid squares). No replication of Ebola.DELTA.VP30-neo was detected in wild-type Vero cells (FIG. 3, bottom panels, solid squares); the low titers that were detected for up to three days postinfection likely reflect input virus. Together, these findings attest to the biological confinement of the Ebola.DELTA.VP30 system. The replication kinetics of Ebola.DELTA.VP30-neo in VeroVP30 cells are similar to those of wild-type Ebolavirus in either VeroVP30 (FIG. 3, top panels, open circles) or wild-type Vero cells (FIG. 3, bottom panels, open circles), establishing the described approach as a highly efficient method for generating biologically contained Ebolaviruses.

[0317] Morphology of Ebola.DELTA.VP30-Neo Virus.

[0318] Next, the morphology of Ebola.DELTA.VP30-neo virus was assessed by transmission electron microscopy (TEM). VeroVP30 cells were infected with Ebola.DELTA.VP30-neo virus and fixed 36 hours later. Samples were processed for TEM as described in Noda et al. (2002). As shown in FIGS. 4A-4B (right panels), the particles budding from VeroVP30 cells infected with Ebola.DELTA.VP30-neo virus were indistinguishable in their size and shape from wild-type Ebolaviruses (FIGS. 4A-4B, left panels). Thus, providing VP30 protein in trans does not have a discernable effect on virion morphology, suggesting that the described system would be suitable for studies of virion formation and budding, for example.

[0319] Taken together, the above results demonstrate that the Ebola.DELTA.VP30-neo virus is biologically contained, replicates to high titers in a helper cell line, is genetically stable, and is morphologically indistinguishable from wild-type virions. Having provided proof-of-concept for the generation of biologically contained Ebolaviruses, the utility of this strategy in basic research and drug screening applications was assessed.

[0320] Generation of an Ebola.DELTA.VP30-eGFP Virus and its Usefulness for Basic Research Applications.

[0321] An Ebola.DELTA.VP30 virus encoding enhanced green fluorescence protein (eGFP) instead of VP30 was generated (FIG. 1; designated Ebola.DELTA.VP30-eGFP), using the same procedures described above for Ebola.DELTA.VP30-neo virus. Analogous to Ebola.DELTA.VP30-neo virus, the eGFP variant replicated efficiently with virus titers reaching 8.0.times.10.sup.7 FFU/mL. Expression of eGFP was observed as early as 10 hours postinfection (data not shown).

[0322] Takada et al. (2003) used replication-competent vesicular stomatitis virus (VSV) pseudotyped with Ebolavirus GP and two neutralizing monoclonal antibodies (mAb), 133/3.16 and 226/8.1, to map Ebolavirus GP epitopes and to generate escape mutants. To confirm with authentic Ebolavirus virions the findings of Takada et al. (2003) based on a VSV-pseudotyping system, escape mutants were generated by amplifying Ebola.DELTA.VP30-eGFP virus in the presence of mAb 133/3.16 or 226/8.1. Each of eight escape mutants to mAb 133/3.16 possessed a histidine-to-arginine substitution at position 549 (H549R) in GP, reported by Takada et al. (2003). Using mAb 226/8.1, 12 escape mutants were isolated that all contained an arginine-to-tryptophan substitution at position 134 (R134W), a mutation identical to one identified by Takada et al. (2003). However, the remaining two escape mutations described by Takada et al. (2003) were not detected. Whether this discrepancy in escape mutants reflects differences between the biological systems used or random mutations is presently unclear. Nonetheless, these experiments illustrated one of the ways that biologically contained Ebolaviruses could be used in basic research applications.

[0323] In conclusion, biologically contained Ebolaviruses lacking the VP30 gene afford a safe, alternative way to study authentic Ebolavirus, to develop Ebolavirus vaccines, and to screen chemical libraries for compounds that interfere with the Ebolavirus life cycle. Indeed, each of the three different biologically contained viruses generated (encoding neomycin or eGFP instead of VP30) was biologically contained, as demonstrated by their ability to replicate in VeroVP30 (a Vero cell line that stably expresses VP30 in trans), but not in wild-type Vero cells. Moreover, virus titers were in the range of 10.sup.7 FFU/mL and hence comparable to those obtained for wild-type Ebolavirus (FIG. 3; Volchov et al., 2001; Neumann et al., 2002; Ebihara et al., 2006) while morphological, biochemical, and virological analyses indicated that the tested properties of Ebola.DELTA.VP30 viruses were indistinguishable from those of wild-type Ebolavirus.

[0324] These physical properties, together with the results of studies to illustrate the potential of biologically contained Ebolaviruses in basic research and drug screening applications, will greatly accelerate current filovirus research efforts.

Example 2

[0325] Ebola viruses (family Filoviridae), cause severe hemorrhagic fever in humans and nonhuman primates with mortality rates up to 90% (Johnson et al., 1977). Currently, there are no licensed vaccines or antivirals available against Ebola virus. A vaccine against Ebola virus is not only desirable for local populations in the epidemic areas of Africa, but also for health care workers during an outbreak and for post-exposure treatment of laboratory workers after accidental exposure to the virus. A few vaccine candidates have been shown to protect mice, guinea pigs, or nonhuman primates against a lethal challenge of Ebola virus; however, each of these candidates has disadvantages, such as lack of protection in nonhuman primates, preexisting immunity against the vector in humans, or potential central nervous system involvement (Reed et al., 2007). Moreover, the current vaccine candidates are based on virus-like particles (VLPs) or virus-vectored vaccines, none of which express the full components of the viral antigens. On the other hand, the use of live attenuated vaccines may not be feasible for Ebola virus from a biosafety perspective. To overcome these potential limitations, biologically contained viruses offer an attractive option since they are biologically safe but provide all the viral antigens.

Materials and Methods

[0326] Cells.

[0327] VeroVP30 cells were established as described in Example 1 and grown in Eagle's minimal essential medium (MEM) supplemented with 10% fetal calf serum (FCS), L-glutamine, vitamins, nonessential amino acid solution, and 5 .mu.g/mL puromycin (Sigma, St. Louis, Mo.).

[0328] Viruses.

[0329] The Ebola.DELTA.VP30 virus was generated as described in Example 1. Briefly, using the plasmid containing the full-length Ebola cDNA genome of the Zaire Mayinga strain of Ebola virus (Neumann et al., 2002), the open reading frame (ORF) of VP30 was replaced with the ORF of the drug-resistant gene neomycin. Using Ebola virus reverse genetics (Neumann et al., 2002), the Ebola.DELTA.VP30 virus was generated and passaged in a Vero cell line stably expressing VP30. Ebola.DELTA.VP30 was propagated in VeroVP30 cells in MEM medium as described above, but supplemented with 2% FCS. The virus was harvested six days after infection of the cells at a multiplicity of infection (MOI) of 1 and directly stored at -80.degree. C. Harvested virus was also partially purified by ultracentrifugation at 27,000 rpm for 2 hours over 20% sucrose. The viral pellet was resuspended in sterile PBS and stored at -80.degree. C. Viral titers were determined by plaque assay in confluent VeroVP30 cells overlaid with 2% FCS-MEM containing 1.5% methyl cellulose (Sigma).

[0330] Since wild-type Ebola virus does not kill mice, challenge studies were carried out with a mouse-adapted Ebola virus (Bray et al., 1998). This virus was generated as described in Ebihara et al., 2006 and used under BSL-4 conditions at the Canadian Centre for Human and Animal Health in Winnipeg, Canada.

[0331] Antibody Titers.

[0332] The levels of Ebola glycoprotein (GP)-specific immunoglobulin G (IgG) antibodies in vaccinated mice were examined by using an enzyme-linked immunosorbent assay (ELISA). Briefly, wells of Immulon 2HB plates (Thermon Labsystems, Franklin, Mass.) were coated with purified Ebola GP (Takada et al., 2001) and blocked with PBS containing 10 mg/mL bovine serum albumin. After incubation of Ebola GP-coated wells with mouse serum from control and vaccinated mice, bound antibodies were detected with goat anti-mouse IgG conjugated to horseradish peroxidase (Kirkegaard & Perry Laboratories Inc., Gaithersburg, Md.) by an ELISA plate reader at an absorbance of 405 nm.

[0333] Intracellular Staining and Flow Cytometry.

[0334] The number of cytokine-producing CD8.sup.+ T cells was determined by intracellular staining as described Murali-Krishna et al. (1998). Briefly, splenocytes were stimulated with the Ebola peptide NP.sub.279-288 (SFKAALSSLA, derived from the nucleoprotein NP; SEQ ID NO: 31) (Olinger et al., 2006; Simmons et al., 2004), VP40.sub.171-180 (YFTFDLTALK, derived from the matrix protein VP40; SEQ ID NO: 32), or GP.sub.161-169 (LYDRLASTV, derived from GP; SEQ ID NO: 33) (Olinger et al., 2005; Warfield et al., 2005) for 5 hours in the presence of brefeldin A and IL-2. Following activation, cells were stained for cell surface CD8.sup.+ and intracellular IFN.gamma. by using the Cytofix/Cytoperm kit from BD Biosciences (San Jose, Calif.). The number of cytokine-producing CD8.sup.+ T cells was determined by using a FACSCalibur flow cytometer (BD Biosciences).

[0335] Vaccination and Challenge.

[0336] Four-week-old female BALB/c mice (The Jackson Laboratory, Bar Harbor, Me.) were anesthetized with isoflurane and intraperitoneally (IP) inoculated twice at three-week intervals with 10.sup.6 focus forming units (FFU) of sucrose-purified Ebola.DELTA.VP30 virus (FIG. 7); control mice were simultaneously inoculated with PBS. A second group of mice received three immunizations (at three-week intervals) with 10.sup.7 FFU of virus harvested from cell culture supernatant (FIG. 7), or, as a control, 2% FCS-MEM. Vaccinations were conducted at the University of Wisconsin-Madison. Mice were then transported to the BSL-4 laboratory at the National Microbiology Laboratory of the Public Health Agency of Canada, where they were challenged with 1000 mouse lethal doses 50 (MLD.sub.50; i.e., the dose required to kill 50% of infected animals) of mouse-adapted Ebola virus. Four days after challenge, viral titers were determined in the serum of three control and three vaccinated mice from each group. The remaining mice were monitored for survival for 28 days. All animal experiments were performed in accordance with approved animal use protocols and according to the guidelines set forth by the Canadian Council of Animal Care and the University of Wisconsin-Madison.

Results

[0337] Antibody Response of Mice Immunized with Ebola.DELTA.VP30 Virus.

[0338] To assess the Ebola.DELTA.VP30 virus as a potential vaccine, its immunogenicity in mice was determined. Mice vaccinated with the Ebola.DELTA.VP30 virus did not show any signs of disease, demonstrating the lack of pathogenicity of the Ebola.DELTA.VP30 virus. When serum samples, collected two weeks after each vaccination to determine the levels of antibodies to the Ebola glycoprotein (GP), were tested for IgG antibody by ELISA with purified GP (FIGS. 5A-5B), vaccinated animals showed elevated levels of antibody titers against GP after the first vaccination compared to control mice (FIGS. 5A-5B); these antibody titers further increased after the second and third vaccinations. This finding demonstrates the ability of the biologically contained Ebola.DELTA.VP30 virus to elicit antibodies to GP.

[0339] CD8.sup.+ T-Cell Responses in Vaccinated Mice.

[0340] The cellular response to vaccination in mice was examined. Mice were vaccinated as described above. Eight days after the second immunization, four vaccinated and two control mice were euthanized and their spleens removed. Splenocytes were isolated and stimulated with the Ebola peptide NP.sub.279-288 (SFKAALSSLA; SEQ ID NO: 31), VP40.sub.171-180 (YFTFDLTALK; SEQ ID NO: 32) or GP.sub.161-169 (LYDRLASTV; SEQ ID NO: 33) for 5 hours in the presence of brefeldin A and IL-2. Vaccinated mice had IFN.gamma.-positive CD8.sup.+ cells in the range of 0.017% to 0.22% for cells stimulated with Ebola peptide NP.sub.279-288 (FIG. 6). For control mice, the number of IFN.gamma.-positive CD8.sup.+ cells was significantly lower, ranging from 0.00513% to 0.00794% (FIG. 6). No IFN.gamma.-positive CD8.sup.+ cells were detected for cells stimulated with Ebola peptide VP40.sub.171-180 or GP.sub.161-169 (data not shown).

[0341] Protective Efficacy of Ebola.DELTA.VP30 Virus in Mice.

[0342] To assess the protective efficacy of the Ebola.DELTA.VP30 virus, two groups of 4-week-old mice were intraperitoneally immunized, then subjected to lethal challenge with mouse-adapted Ebola virus (FIG. 7). `Group 1` mice were immunized three times at three-week intervals with 10.sup.7 FFU of non-purified Ebola.DELTA.VP30 virus (i.e., virus harvested from cell culture supernatant); eight control mice were inoculated in the same manner with 2% FCS-MEM. Mice from this group were challenged seven weeks after the last immunization with 1000 MLD.sub.50 of mouse-adapted Ebola virus, which consistently kills mice (Bray et al., 1998; Ebihara et al., 2006). `Group 2` mice were immunized twice (with a three-week interval) with 10.sup.6 FFU of purified Ebola.DELTA.VP30 virus; ten control mice were similarly inoculated with PBS. Mice from `Group 2` were challenged eight weeks after the last immunization with 1000 MLD.sub.50 of mouse-adapted Ebola virus. No signs of disease or illness were seen in mice vaccinated with purified or non-purified Ebola.DELTA.VP30 virus, whereas control mice from both groups began showing signs of sickness (e.g., ruffled fur) along with weight loss on day 3 post-challenge (FIG. 8A). By day 7 post-challenge, all control mice had succumbed to infection (FIG. 8B). By contrast, vaccinated mice from both groups showed no signs of disease, as characterized by ruffled fur and weight loss (FIG. 8A), and were fully protected against lethal challenge (FIG. 8B) up to day 28, when all surviving mice were euthanized. On day 4 post-challenge, mice were sacrificed to determine viral titers in the sera (FIG. 9). Vaccinated mice from both groups showed a 3 to 4 log.sub.10 reduction in viral titers compared to their respective control mice. Taken together, these data demonstrate that the Ebola.DELTA.VP30 virus efficiently protects mice against challenge with a lethal dose of mouse-adapted Ebola virus.

Discussion

[0343] Here, it was demonstrated that Ebola.DELTA.VP30-immunized mice were completely protected from a lethal challenge with mouse-adapted Ebola virus and that the virus titers in sera from these mice were more than 1000-fold lower than those in control mice. These results show the potential of this biologically contained Ebola virus as a vaccine.

[0344] The humoral response to Ebola virus infection is important, as demonstrated by protection from a lethal challenge by passive transfer of antibodies to the viral glycoprotein GP (Gupta et al., 2001; Warfield et al., 2003). However, the ability of a vaccine to elicit an antibody response does not in itself correlate with protection from Ebola virus infection. For example, classical vaccine approaches, such as .gamma.-irradiated Ebola and Marburg viruses, along with GP expressed in baculovirus generate a moderate antibody response; however, they fail to protect mice against a lethal challenge (Ignatyeve et al., 1996; Lupton et al., 1980; Mellquist-Riemenschneider et al., 2003). By contrast, Ebola and Marburg VLPs protect mice from a lethal challenge of Ebola or Marburg virus (Warfield et al., 2003; Warfield et al., 2004; Warfield et al., 2005), and not only elicit a humoral response, but also induce a CD8.sup.+ T-cell response, highlighting the importance of the latter response for protection against a lethal challenge of Ebola virus (Warfield et al., 2005). Similarly, in non-human primates (NHPs), full protection from a lethal challenge appears to depend on both the humoral response and a CD8.sup.+ cellular response (Sullivan et al., 2000). Vaccine candidates that protect NHPs from a lethal Ebola virus challenge, such as recombinant vesicular stomatitis virus (VSV) (Jones et al., 2005) and adenovirus (Sullivan et al., 2000), induce a CD8.sup.+ T-cell response in NHPs, albeit to varying degrees (Jones et al., 2005; Sullivan et al., 2000). The Ebola.DELTA.VP30 virus induced both humoral and CD8.sup.+ T-cell (specific for an Ebola NP epitope) responses, although the extent of the latter responses varied among animals (FIG. 6). Whether this CD8.sup.+ T-cell response is sufficient to provide protection to NHPs from a lethal Ebola virus infection remains to be tested.

[0345] Although vaccine candidates such as recombinant VSV or parainfluenza virus offer protection in various animal models (Bukreyev et al., 2006; Jones et al., 2005), there are safety concerns with the use of these vaccines in humans (Bukreyev et al., 2006; Jones et al., 2005; Reed et al., 2007). Preexisting immunity to a vaccine based on recombinant adenovirus is also a concern, as is the large amount of virus (10.sup.10 particles) needed to confer protection in NHPs (Jones et al., 2005; Sullivan et al., 2000). Ebola and Marburg VLPs have been shown to protect mice and guinea pigs from a lethal challenge of these viruses (Warfield et al., 2004; Warfield et al., 2005). While VLPs are safe and, due to the rarity of Ebola virus infection, preexisting immunity to Ebola or Marburg viruses is not a concern for VLP vaccines, it is difficult to produce large quantities of VLPs from cell culture.

[0346] The biologically contained Ebola.DELTA.VP30 virus is thus an ideal vaccine candidate since it combines the advantages of VLPs and vectored vaccines (i.e., safety and efficacy), yet it can be propagated to high titers in VeroVP30 cells like standard viruses (Example 1). Further studies will include testing the Ebola.DELTA.VP30 virus for its protective efficacy in NHPs. In addition, shorter, single vaccination protocols will be evaluated to determine if the Ebola.DELTA.VP30 virus vaccine could elicit fast and effective immunity in the event of an outbreak or bioterrorism attack. This includes evaluating the Ebola.DELTA.VP30 virus as a vaccine for post-exposure treatment.

Example 3

Generation of Noninfectious Ebola Particles

[0347] Materials and Methods

[0348] Cells.

[0349] 293 and 293T human embryonic kidney cells were maintained in DMEM supplemented with 10% fetal calf serum, 2% L-glutamine, and penicillin-streptomycin solution (DMEM-FCS) (Sigma). The cells were grown at 37EC in 5% CO.sub.2.

[0350] Construction of Plasmids.

[0351] To generate cDNA constructs encoding the VP40 protein, primers were used that bind to the start and stop codons (positions 4479 and 5459 of the positive-sense antigenomic RNA) to reverse transcribe and PCR-amplify purified viral RNA (Titan RT-PCR Kit, Roche). The PCR product was cloned in the pT7Blue vector (Novagen) resulting in pT7EboZVP40. The cloned Ebola VP40 gene was sequenced to ensure that unwanted nucleotide replacements were not present.

[0352] To generate plasmid pETEBoZVP40His for the expression of 6-histidine-tagged VP40 in Escherichia coli, pT7EboZVP40 was used as a template for PCR amplification with the appropriate primers. The PCR product was blunt-end ligated into the SmaI-digested site of vector pM (CLONETECH). This construct was digested with NdeI and EcoRI and the fragment containing VP40 was ligated into the expression vector pET-5a (Promega). To generate plasmids pCEboZVP40, pCEboZVP40AAXY, pCEboZVP40M 14A, pCEboZVP40/1-276, pCEboZVP40/1-226, pCEboZVP40/1-176, pCEboZVP40/50-326, and pCEboZVP40/100-326 (proteins expressed from these plasmids are designated VP40, VP40AAXY, and the like) for expression of VP40 and its mutants in eukaryotic cells, the Ebola Zaire VP40 gene was amplified from pT7EboZVP40 using specific forward primers, each containing an EcoRI site 5' to the start of the coding region, and specific reverse primers, each containing a Bglll site 3' to the stop codon for each construct, and blunt-end ligated into the EcoRV-digested site of vector pT7Blue. Each construct was digested with EcoRI and Bglll, and the fragment containing the VP40 gene or modified VP40 gene was cloned into the EcoRI and BglII-digested eukaryotic expression vector pCAGGS/MCS (expression controlled by the chicken .beta.-actin promoter) (Kobasa et al., 1997; and Niwa et al., 1991).

[0353] Antibody.

[0354] A polyclonal antibody against Ebola Zaire VP40 was produced as follows: BL21 E. coli cells were transformed with plasmid pETEboZVP40His. Expression of the 6-His-tagged VP40 protein was induced with 1 mM IPTG for 3 hours. The E. coli cells were lysed and cellular debris was remove by centrifugation. The supernatant was purified over an Ni-NTA agarose column (Qiagen). Expression of VP40 was verified by SDS-PAGE followed by Western blotting using a monoclonal antibody against the histidine tag (Kodak). Rabbits were immunized with approximately 0.5 mg of VP40, and antibody against keratin present in the antiserum was removed with a keratin column (Girault et al., 1989).

[0355] Cell Transfection for Expression of VP40 and its Mutants.

[0356] 293 or 293T cells (60-mm plates) were transfected with expression vectors with the use of the Trans IT LT-1 liposomal reagent (Panvera) according to the manufacturer's instructions. Briefly, DNA and transfection reagent were mixed (6 .mu.L of Trans IT LT-1 with 3 .mu.g of DNA) in 0.2 mL OPTI-MEM (Gibco-BRL), incubated for 30 minutes at room temperature, and added to the cells. Transfected cells were incubated at 37EC until harvest of the supernatant and/or cell monolayer.

[0357] Particle Formation Assay.

[0358] Particles were assayed by the method of Li et al (1993) with some modifications. Forty-eight hours after transfection of 293T cells with pCEboZVP40, pCEboZVP40AAXY, pCEboZVP40M14A, or pCEboZVP40/1-276, the culture medium was removed and placed on ice. The cell monolayer was washed with phosphate-buffered saline (PBS), scraped into lysis buffer (0.25 M Tris-HCl, pH 8.0, 0.5% Triton X-100) and kept at 4EC. The culture medium (2 mL) was centrifuged at 2,000 rpm in a microcentrifuge for 5 minutes to remove cellular debris, layered over 20% sucrose in STE buffer (0.01 M Tris-Cl, pH 7.5, 0.01 M NaCl, 0.001 M EDTA, pH 8.0) (2 ml), and centrifuged at 150,000.times.g for 2 hours at 4EC. After centrifugation, the supernatant was removed and added to the cell lysate. This mixture was saved for analysis of total protein expression. The pellet was resuspended in 1 mL STE buffer overnight at 4EC. The resuspended pellet was layered over a 10-50% discontinuous sucrose gradient in STE buffer, centrifuged at 150,000.times.g for 4 hours at 4EC, and fractions (1 mL) were collected from the top of the gradient. Each fraction was mixed with 0.25 ml of 50% trichloroacetic acid (TCA) (10% TCA), the fractions were incubated for 30 minutes on ice, and the precipitated proteins were pelleted by microcentrifugation for 15 minutes. The pellets were washed once with cold acetone, air-dried, and resuspended in 0.05 ml SDS-PAGE sample buffer. Proteins in the mixture of cell lysate and supernatant from centrifugation through 20% sucrose were precipitated with 10% TCA, washed with acetone, and resuspended in 0.5 mL SDS-PAGE sample buffer. Proteins were separated by 12% SDS-PAGE and detected by Western blotting. Fractions are numbered from the top to the bottom of the gradient.

[0359] Protease Protection Assay.

[0360] 293T cells were transfected with pCEboZVP40 and, at 48 hours post-transfection, the culture medium was removed. The medium was microcentrifuged at 2,000 rpm for 5 minutes to remove cellular debris, layered over a 20% sucrose cushion, and centrifuged at 165,000.times.g for 1 hour at 4EC. The supernatant was removed and the pellet was resuspended overnight at 4EC in 0.4 mL STE buffer. This resuspension was divided into six aliquots and treated following a protocol previously described (Mik et al., 1989): Aliquot 1 received no further treatment; aliquot 2 was treated with soybean trypsin inhibitor (Biofluids) to a final concentration of 3 mg/ml; aliquot 3 with triton X-100 to a final concentration of 1%; aliquot 4 with trypsin (Worthington) to a final concentration of 0.1 mg/mL; aliquot 5 with both Triton X-100 to 1% and trypsin to 0.1 mg/ml final concentration; and aliquot 6 with both trypsin inhibitor (3 mg/ml final) and trypsin (0.1 mg/mL final). The samples were incubated at room temperature for 30 minutes, after which an excess of trypsin inhibitor (5 mg/mL) was added to each aliquot. SDS-PAGE sample buffer (6.times.) was added to each aliquot. Proteins from each aliquot were separated by 12% SDS-PAGE and detected by Western blotting.

[0361] Membrane-Association Assay.

[0362] The method of Bergmann and Fusco (1988) was used, with some modifications, to determine membrane-association of VP40 and its mutants. Briefly, 48 hours after transfection of 293 cells with pCEboZVP40 or a mutant-VP40 expression plasmid, the culture medium was removed, and the cell monolayer, after a wash with (PBS), was scraped into ice-cold sucrose homogenization buffer (10% wt/wt sucrose, 10 mM Tris-HCl (pH 7.4), 1 mM EDTA, and 10 mM iodoacetamide). Cells were disrupted with 30 strokes of a Dounce homogenizer on ice and microcentrifuged for 3 minutes at 2,000 rpm to remove nuclei. The resulting supernatant was made to 1 M NaCl or left untreated, incubated at room temperature for 20 minutes, made to 80% sucrose (wt/vol), placed at the bottom of a Beckman SW41 centrifuge tube, and overlaid with 5 ml of 65% (wt/vol) sucrose and 2.5 mL of 10% sucrose. The gradient was centrifuged to equilibrium at 150,000.times.g for 18 hours at 4EC. Fractions (1 mL) were collected from the top of the gradient, diluted 1:1 with TBS-Triton buffer (0.025 M Tris-HCl, pH 7.5, 0.15 M NaCl, 0.5% Triton X-100) or, for experiments involving expression of VP40/100-326, precipitated with TCA (as described for the particle formation assay) owing to the weak signal of this deletion construct in Western analysis, and mixed with SDS-PAGE sample buffer. Proteins from each aliquot were separated by 12% SDS-PAGE and detected by Western blotting.

[0363] Triton X-114 Phase Partitioning Analysis.

[0364] The method used was essentially that of Bordier (1981). Forty-eight hours post-transfection of 293 cells pCEboZY40, pCEboZVP40/1-276, pCEboZVP40/1-226, pCEboZVP40/1-176, pCEboZP40/50-326, pCEboZVP40/100-326, or, as a control, a vector expressing ANVSN/33 (H1N1) influenza virus hemagglutinin (HA), cells were scraped into cold TN buffer (10 mM Tris-HCl, pH 7.4, 150 mM NaCl), disrupted with 30 strokes in a Dounce homogenizer, and subjected to centrifugation at 2,000 rpm for 3 minutes to remove nuclei. Triton X-114 (Sigma) was added to each supernatant to 1%, and the resulting solution was incubated for 15 minutes at 4EC with agitation. Unsolubilized material was pelleted by centrifugation in a picofuge for 5 minutes at 4EC, and the supernatant was heated to 37EC for 5 minutes. The supernatant was layered onto a 37EC sucrose (6%) cushion in TN buffer containing 0.06% Triton X-114 and centrifuged at 2,000 rpm for 3 minutes at room temperature. The detergent (lower) and aqueous (upper) phases were recovered separately, the aqueous phase was extracted a second time, like phases were pooled, and the detergent phase was diluted in TN buffer. Proteins in each phase were precipitated with 50% acetone and resuspended in SDS-PAGE sample buffer. Proteins were separated by 12% SDS-PAGE and analyzed by Western blotting.

[0365] Western Blotting.

[0366] Samples in sample buffer (10 .mu.L) were incubated at 100EC for 5 minutes and separated on 12% polyacrylamide gels. Resolved proteins were transferred to Westran polyvinylidine difluoride membranes (Schleicher & Schuell) and blocked overnight at 4EC with 5% skim milk in PBST (0.05% Tween 20 (Sigma) in PBS). Blots were incubated in primary antibody for 1 hour at room temperature, washed three times with PBST, incubated in biotinylated anti-rabbit secondary antibody (Vector Laboratories) for 30 minutes, washed three times with PBST, incubated in streptavidin-horseradish peroxidase reagent (Vector Laboratories) for 30 minutes and washed three times with PBST. Blots were then incubated in Lumi-Light Western blotting substrate (Boehringer-Mannheim) for minutes and exposed to x-ray film (Kodak).

Results

[0367] Expression of VP40 in Mammalian Cells.

[0368] To ensure that VP40 is expressed at efficient levels in human embryonic kidney 293T cells, the cell lysate was analyzed 24 hours after transfection with pCEboZVP40 by Western blotting. Two bands reacting with anti-VP40 polyclonal antibody were found, a small distance apart, in the range of 40 kDa. The lysate from cells transfected with the expression vector alone did not react with the antibody.

[0369] VP40 contains an internal start codon at nucleotides 40-42 (codon 14) that is in frame with the first AUG. To determine whether protein synthesis from this internal start codon was responsible for the faster-migrating band on the gel, a construct was generated, pCEboZVP40M14A, which expresses a mutant VP40 with this second AUG changed to GCG, which encodes alanine, and expressed it as described above. Analysis of the cell lysate revealed a single, larger-sized band, suggesting that the second AUG is used as a start codon to an appreciable extent in this system.

[0370] To determine whether loss of the PPXY motif at amino acids 10-13 of VP40 affects expression of the protein, 293T cells were transfected with pCEboZVP40AAXY, which expresses a mutant VP40 in which the PPEY sequence at amino acids 10-13 was changed to AAEY. Two bands corresponding to those seen with the expression of wild-type VP40 were detected. However, in contrast to the results obtained with wild-type VP40 expression, where the slower-migrating band was the predominate product, pCEboZVP40AAXY expressed the two products at similar levels, indicating that loss of the PPXY motif affects either the translation of VP40 or its stability.

[0371] Production of Membrane-Bound Particles.

[0372] To determine whether VP40-associated vesicles are produced when the protein is expressed in the absence of other viral proteins, 293T cells were transfected with pCEboZVP40 and, after 48 hours, collected the supernatant. After removal of cellular debris, the supernatant was subjected to ultracentrifugation over a 20% sucrose cushion. The pellet was resuspended and centrifuged through a 10-50% discontinuous sucrose gradient, and fractions were analyzed by Western blotting. Fractions 6-8 contained VP40, with the majority of the protein found in fraction 7. The VP40 in fractions 6-8 was most likely associated with membrane lipids in a particle-like structure, as the sucrose densities in these fractions ranged from 1.11 to 1.13 g/mL, which corresponds to findings for matrix protein-generated particles of other viruses (Giddings et al., 1998; Sandefur et al., 1998). Bands detected below full-length protein in the total protein fraction are likely degradation products. These data indicate that VP40 expressed in the absence of other viral proteins can produce membrane-bound particles.

[0373] Protease Protection Assay.

[0374] To confirm the ability of VP40 to produce membrane-bound particles when expressed alone, a trypsin protection assay was employed. Culture supernatant from cells transfected with pCEboZVP40 was centrifuged at 165,000.times.g through 20% sucrose, and the pellet was resuspended in STE buffer and divided into six equal aliquots. Aliquots 1-3 served as controls (untreated, trypsin inhibitor treated, and triton X-100 treated), aliquot 4 was treated with trypsin, aliquot 5 with trypsin and triton X-100, and aliquot 6 with trypsin inhibitor and trypsin. Trypsin degraded VP40 only in the presence of triton X-100, indicating that the viral protein does induce the production of fully membrane-bound particles; that is, trypsin digestion of VP40 required disruption of the lipid-bilayer surrounding the protein.

[0375] VP40 Mutants and Membrane-Bound Particle Formation.

[0376] Does the PPXY motif at amino acids 10-13 of VP40 contribute to particle production? To address this question, VP40AAXY was expressed in 293T cells and assayed for particles as described for wild-type VP40. VP40AAXY was not detected in fractions corresponding to the sucrose densities to which wild-type VP40 particles migrated. Since VP40AAXY was synthesized at levels similar to wild-type VP40, this finding indicates that mutation of the PPXY motif markedly disrupts VP40-generated vesicle formation.

[0377] A substantial amount of VP40M14A was present in fractions 5-8 in the gradient, and the percentage of total VP40M14A expressed in 293T cells that contributed to membrane-bound particle formation was much greater than the percentage of total wild-type VP40 involved in particle formation. This result is consistent with the finding that the PPXY motif present immediately upstream of the second AUG is critical for VP40-associated particle formation.

[0378] To determine whether the C-terminus of VP40 is essential for particle formation, a deletion mutant, VP40/1-276, was assayed which lacks the final 50 amino acids of VP40, for particle generation. Since this deletion mutant was not present at the same sucrose densities that characterized the migration of wild-type VP40, it was concluded that the first 276 amino acids of VP40 are not sufficient for particle formation.

[0379] VP40 Association with Cell Membranes and Structural Requirements for Activity.

[0380] Flotation analysis was used to determine if VP40 binds cellular membranes efficiently in mammalian cells. In this method, postnuclear membrane fractions in 80% sucrose are loaded at the bottom of a centrifuge tube and overlaid with 65% and 10% sucrose. During centrifugation, cellular membranes and their associated proteins float to the 10-65% sucrose interface, while soluble proteins remain in the dense sucrose fractions at the bottom of the tube.

[0381] A large percentage of wild-type VP40 was found at the 10-65% sucrose interface (fraction 3), while the remaining protein was found in the loading zone (fractions 8-12), indicating that VP40 does indeed bind cellular membranes. To clarify the interactions involved in this association, VP40-associated membranes were treated with 1 M NaCl to determine whether electrostatic interactions were required for this association and subjected them to flotation analysis. Salt treatment had a negligible affect on the ability of VP40 to associate with membranes, suggesting that the protein contains at least one hydrophobic domain able to associate with membranes.

[0382] To elucidate the domain(s) of VP40 important for membrane association, deletion mutants were generated. Constructs expressing amino acids 50-326 (pCEboZVP40/50-326), amino acids 100-326 (pCEboZVP40/100-326), amino acids 1-176 (pCEboZVP40/1-176), amino acids 1-226 (pCEboZVP40/1-226), and amino acids 1-276 (pCEboZVP40/1-276) of VP40 were expressed in 293 cells and their membrane association in the presence or absence of 1 M NaCl was examined. The mutants with the largest truncations, VP40/1-176 and VP40/100-326, showed the highest level of association with the lipid bilayer. Salt treatment did not affect these interactions. Mutants VP40/1-226 and VP40/50-326 associated with membranes to the extent found with wild-type VP40, and these interactions were also relatively unperturbed by treatment with salt. By contrast, only a small portion of VP40/1-276 associated with the lipid bilayer, and this interaction was eliminated upon treatment with salt. These results indicate that loss of the C-terminal 50 amino acids of VP40 markedly alters the membrane-binding capabilities of VP40, primarily by disrupting hydrophobic interactions. This effect was ameliorated when 50 additional C-terminal amino acids were deleted, and membrane-association was promoted when the protein was further truncated to 176 amino acids. Deletion of the N-terminal 49 amino acids of VP40 did not alter the membrane-binding characteristics of the protein, although truncation of 50 additional N-terminal amino acids did enhance protein-membrane association, as seen with VP40/1-176.

[0383] Since particle formation was markedly reduced with VP40AAXY, cells expressing this mutant were subjected to flotation analysis in order to determine whether a decreased ability to bind membranes was involved in this deficiency. The loss of the PPXY motif in VP40 did not affect the ability of the protein to bind membranes, indicating that lack of particle production with this mutant was not due to the loss of membrane association.

[0384] Flotation analysis was also used to determine whether the more efficient particle formation induced by VP40M14A, by comparison to wild-type VP40, could be attributed, at least in part, to increased membrane binding by this mutant. The percentage of VP40M14A associated with membranes was only slightly greater than that determined for wild-type VP40, indicating that this mutant relies on another mechanism to increase particle formation.

[0385] Triton X-114 Phase Partitioning Analysis.

[0386] To probe the nature of the VP40-membrane interaction further, Triton X-114 phase partitioning analysis was used as integral membrane proteins and lipid anchored proteins partition in the detergent phase of a protein extraction and peripheral membrane proteins partition in the aqueous phase. HA, an integral membrane protein, was found entirely in the detergent phase of the extraction, as expected. Only a small portion of total VP40 was found in the detergent phase, while VP40/1-276 was found almost entirely in the aqueous phase. VP40/1-226 and VP40/50-326 partitioned in the detergent phase in proportions similar to that found for wild-type VP40. By contrast, when VP40/1-176 and VP40/100-326 were expressed, large proportions of each partitioned in the detergent phase. These results indicate that wild-type VP40 possesses only minor traits of an integral membrane protein, and that deletion of its C-terminal 50 amino acids (VP40/1-276) abrogates these features. Further truncation of the C-terminus (VP40/1-226 and VP40/1-176) enhances the integral membrane character of protein. Deletion of the N-terminal 49 amino acids of VP40 (VP40/50-326) does not alter the general structural features of the protein, while deletion of amino acids 1-99 (VP40/100-326) appears to increase the extent of anchoring to lipids.

Discussion

[0387] Thus, VP40 of Ebola virus, when expressed in the absence of other viral proteins, can induce the formation of membrane-encompassed particles, much in the manner of the matrix proteins of VSV, rabies, and simian immunodeficiency virus (Giddings et al., 1998; Harty et al., 1999; Justice et al., 1995; Li et al., 1993). Cellular proteins containing the WW domain are, in all likelihood, crucial for this process, as VP40 containing an altered version of a PPXY motif at amino acids 10-13 induces little or no particle formation. Harty et al. (1999) demonstrated that the matrix proteins of VSV and rabies viruses, which possess this motif at their N-termini, bind the cellular Yes-kinase-associated and Nedd4 proteins via a PPXY motif-WW domain, interaction, and that the loss of this motif results in impaired virus release from infected cells. Jayakar et al. (2000) recently demonstrated that mutation of the PPXY motif in the matrix protein of VSV impedes budding of fully assembled virions at the plasma membrane. The data described herein provides evidence for an important role of the PPXY motif in particle formation induced by VP40, and suggest that cellular proteins are crucial players in this process.

[0388] The efficiency of particle production markedly increased when the second ATG codon of VP40 (codon 14) was changed to GCG (alanine), but the reason for this enhancement remains unclear. This ATG codon immediately follows the PPXY motif. Perhaps the faster-migrating version of VP40, which lacks the PPXY motif, interferes with the assembly or budding of full-length VP40 molecules at the cell surface, or with the interaction between VP40 and a cellular protein. Whether translation from this second ATG occurs in actual viral infection or is an artifact of the system employed in this study is unknown.

[0389] Ruigrok et al. (2000) reported that VP40 expressed in E. coli can bind liposomes in vitro and that this interaction is largely electrostatic. In mammalian cells, a substantial amount of VP40 bound to the cellular membrane, and that this interaction was disrupted negligibly by the presence of 1 M NaCl, indicating that at least one hydrophobic domain is involved in this interaction. A small but appreciable portion of VP40 partitioned with detergent in the manner of an integral membrane or lipid-anchored protein in Triton X-114 phase-partitioning analysis. This result, together with the inability of 1 M NaCl to dissociate VP40 from the lipid bilayer, indicates that the protein has certain properties of an integral membrane protein, as do a number of matrix proteins of negative-stranded RNA viruses (Chong et al., 1993; Zhang et al., 1996), even though Ebola VP40 does not appear to contain a region of significant length and hydrophobicity to span the cell membrane. Short hydrophobic stretches of VP40 may be able to penetrate the lipid bilayer to some extent, lending modest integral-membrane character to the protein.

[0390] Ruigrok et al. (2000) also reported that a deletion mutant of VP40 containing amino acids 31-212 failed to bind liposomes efficiently, indicating that the C-terminus of VP40 is absolutely required for membrane binding. To elucidate the domains involved in the association of VP40 with cellular membranes, carboxy and amino-terminal deletion mutants were constructed. VP40 lacking its C-terminal 50 amino acids demonstrated appreciably reduced membrane association. The Kyte-Doolittle hydrophobicity plot (1982) of VP40 indicates that amino acids 277-326 of the protein are primarily hydrophobic, so that deletion of amino acids 277-326 eliminates a substantial hydrophobic region that is likely important for efficient membrane-binding by the full-length protein. This hypothesis is supported by the fact that 1 M NaCl completely disrupted this association, suggesting that affinity of this deletion construct with the lipid bilayer depends primarily on electrostatic interactions.

[0391] When amino acids 227-326 of VP40 were deleted, the resulting truncated protein associated with the lipid bilayer as efficiently as wild-type VP40; moreover, C-terminal deletion of amino acids 177-326 resulted in a protein with much higher affinity for the lipid bilayer than was found for wild-type VP40. Salt treatment did not perturb membrane association of these truncated versions of VP40, indicating the presence of hydrophobic interactions mediated by the N-terminal 176 amino acids of the protein.

[0392] The hydrophobicity plot indicates that amino acids 227-276, and particularly amino acids 177-226, are primarily hydrophilic. Deletion of the hydrophilic residues present in this region of VP40 may allow the truncated protein to fold into a structure capable of strong hydrophobic association with the cell membrane, perhaps by effectively exposing the highly hydrophobic central domain of the protein. These results are consistent with data obtained by Triton X-114 extraction analysis. Since VP40 lacking its C-terminal 50 amino acids was unable to produce particles, and these C-terminal residues appear to be required for efficient membrane association of VP40, binding of this highly hydrophobic region to the lipid bilayer may be an essential step in the particle formation process.

[0393] The crystal structure of amino acids 31-326 of Ebola virus was recently elucidated by Dessen et al. (2000). It shows VP40 to be distinct from other viral matrix proteins, in that it consists of two similar domains connected by a flexible linker at amino acids 195-200. Ruigrok et al. (2000) showed that amino acids 31-212 of VP40 form hexamers spontaneously in solution. Dessen and associates postulate that, during the life cycle of Ebola virus, VP40 molecules associate with the lipid bilayer through interactions contributed primarily by their C-termini. After membrane binding, the molecules undergo a conformational change that frees their N-termini for hexamerization. These hexamers then form building blocks for a lattice that underlies the plasma membrane, and subsequently may interact with the cytoplasmic tails of viral glycoproteins and/or the ribonucleoprotein complex. This model is based on data demonstrating the hexamerization of VP40 molecules that lack their N-terminal 30 amino acids as well as their C-terminal 114 amino acids. The PPXY motif that appears crucial for membrane-bound particle formation is located at amino acids 10-13 of VP40, and this motif most likely interacts with a cellular protein that exhibits a WW domain during virus particle assembly or budding. It has not yet been demonstrated that VP40 with a truncated C-terminus can form hexamers when the entire N-terminus is present. If hexamerization does occur during virion morphogenesis, the 18 hexamers that form presumably must leave the PPXY motif accessible to cellular proteins that participate in particle formation and/or budding.

Example 4

Particles Comprising Filovirus Matrix Protein and Glycoprotein

Materials and Methods

[0394] Cells.

[0395] 293T human embryonic kidney cells were maintained in Dulbecco's modified Eagle medium supplemented with 10% fetal calf serum, L-glutamine and penicillin-streptomycin-gentamicin solution. The cells were grown in an incubator at 37EC in 5% CO.sub.2.

[0396] Plasmids.

[0397] Full-length cDNAs encoding the Ebola virus (species Zaire) VP40 or GP were cloned separately into a mammalian expression vector, pCAGGS/MCS (Kobasa et al., 1997; Niwa et al., 1991), which contains the chicken .beta.-actin promoter. The resultant constructs were designated pCEboZVP40 and pCEboZGP, respectively.

[0398] Cell Transfection for Expression of VP40 and GP.

[0399] 293T cells (1.times.10.sup.6) were transfected with plasmids using the Trans IT LT-1 reagent (Panvera, Madison, Wis.) according to the manufacturer's instructions. Briefly, 1 .mu.g of DNA in 0.1 mL Opti-MEM (Gibco-BRL) and 3 .mu.L of the transfection reagent were mixed, incubated for 10 minutes at room temperature, and added to the cells. Transfected cells were incubated at 37EC for 24 or 48 hours.

[0400] Electron Microscopy.

[0401] Ultrathin section electron microscopy was performed as follows. Twenty-four hours post-transfection of 293T cells with plasmids, the cells were washed with phosphate-buffered saline (PBS) and fixed for 20 minutes with 2.5% glutaraldehyde (GLA) in 0.1 M cacodylate buffer (pH 7.4). They were scraped off the dish, pelleted by low-speed centrifugation and then fixed for 30 minutes with the same fixative. Small pieces of fixed pellet were washed with the same buffer, postfixed with 2% osmium tetroxide in the same buffer for 1 hour at 4EC, dehydrated with a series of ethanol gradients followed by propylene oxide, embedded in Epon 812 Resin mixture (TAAB) and polymerized at 70EC for 2 days. For immune electron microscopy, cells were fixed with 4% paraformaldehyde and 0.1% GLA, dehydrated and embedded in LR White Resin (London Resin Company Ltd.). Thin sections were stained with uranil acetate and lead citrate, and examined with a JEM-1200EX electron microscope at 80 Kv.

[0402] For negative staining, culture media of 293T cells were collected at 24 hours post-transfection onto a Formvar-coated copper grid, stained with 2% phosphotungstic acid solution (PTA) and examined with a JEM-1200 electron microscope at 80 Kv.

[0403] For immune electron microscopy, the samples were absorbed to Formvar-coated nickel grids and washed with PBS containing 0.5% bovine serum albumin (PBS-BSA). The grids were then treated with mouse anti-GP monoclonal antibody (a mixture of ZGP12, ZGP42, and ZGP133 (31); 1:150 in PBS-BSA) or rabbit anti-VP40 polyclonal antibody (1:300 in PBS-BSA), and rinsed six times with PBS, followed by incubation with a goat antimouse immunoglobulin conjugated to 15-nm gold particles (1:50 dilution; BBInternational) or a goat antirabbit immunoglobulin conjugated to 5-nm gold particles (1:100 dilution; BBInternational). After washing, the samples were fixed for 10 min in 2% glutaraldehyde and negatively stained with 2% PTA.

Results

[0404] Pleomorphic Particle Formation by GP.

[0405] To determine the morphology of vesicles induced by Ebola virus GP expression, GP-expressing cells and their supernatants were analyzed by electron microscopy. The ultrathin sections of these cells showed particle-like structures with surface spikes budding from the plasma membrane; no such structures were observed using cells transfected with the expression vector alone. As previously observed in the recombinant vaccinia virus system (Volchkov et al., 1998), pleomorphic structures similar to virosomes with a range of diameters were apparent in the supernatants of GP-expressing cells. The spikes on the surface of the vesicles reacted with anti-GP monoclonal antibodies, confirming the GP derivation of the structures.

[0406] VP40 Induces Filamentous Particle Formation.

[0407] To determine how VP40 protein expressed in 293T cells is released into culture medium (Harty et al., 2000; Timmins et al., 2001), the VP40-expressing cells were analyzed by transmission electron microscopy. The ultrathin sections of the cells expressing VP40 showed budding of filamentous structures (approximately 65 nm in diameter) on the cell surface. In some cells, the plasma membranes appeared ruffled and to consist of two bilayers. Aggregated ribosomes were occasionally found in the cytoplasm of cells expressing VP40, as were electron-dense filamentous structures (approximately 45 nm in diameter), which were never seen in cells transfected with the expression vector alone. The budding particles and membrane ruffles reacted with rabbit anti-VP40 polyclonal antibody, confirming that VP40 had contributed to the generation of these structures. In studies to further determine the size and morphology of the VP40 particles released from cells, the supernatants of cells expressing this protein were centrifuged through 20% sucrose, and the pelleted material was negatively stained with 2% PTA and analyzed by electron microscopy. Filamentous particles, which had uniform diameters of approximately 65 nm but varied lengths, were observed. These results indicate that VP40 alone can induce the formation of filamentous particles, which bud from the cell surface.

[0408] VP40-GP Interaction in Particle Morphogenesis.

[0409] To determine how GP expression affects VP40-driven particle formation, 293T cells were transfected with both VP40- and GP-expressing plasmids. In ultrathin sections of the transfected cells, filamentous particle-like structures of 80-nm external diameter were observed that were budding from the plasma membrane. The structures possessed spikes of approximately 10 nm on their surface, in contrast to the structures observed in cells expressing VP40 alone. Also, unlike the findings with expression of GP alone, few pleomorphic particles were observed. The particle structures were studied in more detail after negative staining of the particles in culture supernatants of cells expressing both VP40 and GP. Filamentous Ebola virus-like particles with surface spikes of approximately 85-nm in external diameter and lengths that ranged to 10 .mu.m were observed. The spikes projected from the particle surface at 5- to 10-nm intervals and were morphologically indistinguishable from those on the Ebola virion surface (Feldmann et al., 1996; Peters et al., 1995). Labeling of the spikes with a mixture of anti-GP monoclonal antibodies conjugated with gold particles confirmed their identity as GP. Furthermore, when treated with 0.03% Triton X-100 and with both the anti-VP40 antibody conjugated to 5-nm gold particles and a mixture of anti-GP monoclonal antibodies conjugated to 15-nm gold particles, the filamentous particles became labeled with both antibodies, demonstrating that the Ebola virus-like particles contained GP as well as VP40 proteins. These results demonstrate GP incorporation into VP40-generated filamentous structures, without affecting filamentous particle formation.

Discussion

[0410] A hallmark of Ebola virus is its filamentous virions as featured in its family name Filoviridae. The shape of enveloped viruses are determined by viral proteins in retroviruses (Campbell et al., 1997; Gay et al., 1998; Joshi et al., 2000) or by both viral RNA length and proteins in VSV (Pattnaik et al., 1991). Because specific interactions among viral components are required for the formation of defined virion shapes, understanding of such interactions can lead to the identification of targets for the development of antiviral compounds.

[0411] As shown herein by electron microscopy, the expression of VP40 in the absence of any other Ebola virus proteins leads to the formation of filamentous particles, which resemble spikeless virions released into the supernatant of cultured Ebola virus-infected cells (Geisbert et al., 1995). Thus, these results suggest that the Ebola virus VP40 possesses structural information necessary and sufficient to induce the formation of filamentous particles, which then bud from the plasma membrane. Interestingly, some filamentous structures were observed in the cytoplasm of cells expressing VP40 as have been found in the cytoplasm of the cells infected with Ebola virus. Similar structures have also been observed in cells expressing the M1 protein of influenza virus or the GAG protein of retrovirus (Delchambre et al., 1989; Gheyson et al., 1989; Gomez-Puertas et al., 2000). However, the tubular structures observed upon expression of influenza virus M1 alone were not seen during normal viral infection or when M1 was coexpressed with other influenza viral proteins. Thus, VP40 may form intracellular filamentous structures by self-aggregation.

[0412] Membrane ruffles containing VP40 protein were observed in some VP40-expressing cells. The M protein of VSV induces similar double-layered membranes at the cell surface when expressed from recombinant Sendai virus (Sakaguchi et al, 1999). IpaC protein secreted by Shigella flexneri has also been linked to large-scale membrane extension in macrophages, including lamellipodia and membrane ruffles (Kuwae et al, 2001; Tran Van Nhieu et al., 1999), while Salmonella typhimurium triggers the formation of host cell membrane ruffles in nonphagocytic cells (Ginocchio et al., 1994; Zhou et al., 1999). These membrane ruffles are thought to result from interactions between the bacterial proteins, including IpaC, and the actin cytoskeletons of host cells (Tran Van Nhieu et al., 1999; Zhou et al., 1999). In Ebola virus-infected cells, host cell plasma membranes proliferate extensively at the peak stage of viral budding (Geisbert et al, 1995), as observed in cells expressing VP40 alone. Thus, VP40 may interact with actin filaments during the assembly or budding of Ebola virus at the cell surface.

[0413] The impact of glycoprotein interaction with the matrix protein on virion morphology differs among viruses. For example, deletion of the cytoplasmic tails of the influenza virus hemagglutinin and neuraminidase alters virus morphology (Jin et al., 1997; Mitnaul et al., 1996), while the characteristic morphology of rabies virus and VSV do not depend on glycoprotein-matrix protein interaction (Mebatsion et al, 1996; Mebatsion et al., 1994; Schnell et al., 1998; Takada et al., 1997). The Ebola virus GP, like VSV-G, was incorporated into filamentous particles without affecting the morphology of the particles. However, such interaction may contribute to the efficiency of budding, as demonstrated by research on VSV (Jayakar et al., 2000; Mebatsion et al., 1999).

[0414] In conclusion, VP40 induces VP40 containing-filamentous particle formation and GP spikes are incorporated into VP40 induced-filamentous particles upon coexpression of GP and VP40, resulting in Ebola virus-like particles.

Example 5

A Method to Screen for Modulators of Viral Transcription or Replication

[0415] To produce viral vectors for an antiviral screening method, vectors were prepared that expressed a rhabdovirus or filovirus protein and a reporter. In one embodiment, a reporter gene replaces rhabdovirus GP sequences in genomic rhabdovirus DNA. In one embodiment, a reporter gene replaces filovirus GP sequences in genomic filoovirus DNA. In one embodiment, viral protein expression vectors useful with the recombinant genomic DNA may include one expressing filovirus GP and optionally one or more vectors expressing one or more of rhabdovirus N, P, M and L. In another embodiment, a reporter gene replaces sequences in genomic filovirus DNA. The Filovirus protein expression vectors, e.g., Marburg virus or Ebola virus vectors, include one or more of the following sequences: sequences for L, NP, VP30 and/or VP35. If more than one vector is employed, the vectors may be physically linked or each vector may be present on an individual plasmid or other, e.g., linear, nucleic acid delivery vehicle.

[0416] To develop an antiviral to Ebolavirus, the entry process, including receptor binding and/or fusion, was targeted. To identify compounds that interfere with these steps in the viral life cycle, a replication-incompetent Vesicular Stomatitis Virus (VSV) was employed that lacks the VSV glycoprotein gene and contains the GFP gene instead. This replication-incompetent VSV was pseudotyped with Ebola GP glycoprotein. This pseudotyped virus infects cells once, resulting in GFP gene expression. In the presence of compounds that interfere with Ebola GP-mediated binding or fusion, reporter gene expression is abrogated. This system was used to screen about 6,300 compounds at The National Screening Laboratory for the Regional Centers of Excellence in Biodefense at Harvard University, Boston, Mass., and 144 compounds were identified that reduced reporter gene expression by more than 90%.

[0417] To verify whether the compounds indeed inhibit Ebolavirus infection, a biologically contained Ebolavirus expressing GFP protein (Ebola.DELTA.VP30-GFP virus) (see Example 2) was employed. 111 of the originally-identified 144 compounds were tested and 24 were identified that reduced the infectivity of the biologically contained Ebolavirus by at least 90% (FIG. 11). For those compounds, the 50% inhibitory concentration (IC.sub.50) and cytotoxic concentration (CC.sub.50) were determined. Benztropine mesylate emerged as a lead candidate.

[0418] Further studies revealed that benztropine mesylate efficiently reduced the infectivity of VSV pseudotyped with GPs of all known subtypes of Ebolavirus (i.e., Zaire, Reston, Sudan, Ivory Coast); the titers of these pseudotyped viruses were reduced by 98-99%. Benztropin mesylate was also effective against VSV pseudotyped with the GP protein of Marburgvirus, although to a lesser extent (reduction of virus titers of about 75%). On the other hand, benztropine mesylate does not affect the growth of viruses such as VSV and influenza virus, indicating the specificity of this compound for Ebolavirus.

[0419] Binding of Ebolavirus to cell surface activates the phosphoinositide-3 (PI3) kinase-Akt pathway. It was determined that benztropine mesylate did not inhibit the phosphoinositide-3 (PI3) kinase-Akt pathway per se. However, benztropine mesylate was found to inhibit infection of VSV pseudotyped with Ebola GP that was bound to cell surfaces at 0-4.degree. C., temperatures that may disrupt endocytosis and vesicle trafficking.

[0420] Benztropine mesylate is a known and commercially available inhibitor of the dopamine transporter and is used to treat the symptoms of Parkinsons's disease and other neurological disorders. Since benztropine mesylate is known to bind to receptors for neurotransmitters, Ebolavirus might utilize these receptors as second receptors for entry. Thus, benztropine mesylate might inhibit binding of Ebolavirus to neurotransmitter receptors, resulting in the inhibition of activation of PI3 kinase-Akt pathway for entry. Alternatively, or in addition to blocking neurotransmitter receptors, benztropine mesylate may inhibit fusion of the virus envelope with the cellular membrane.

Example 6

Materials and Methods

[0421] Cells.

[0422] VeroVP30 cells were established as previously described (Halfmann et al., 2008) and grown in Eagle's minimal essential medium (MEM) supplemented with 10% fetal calf serum (FCS). Vero and CV-1 cells were cultured under the same conditions as VeroVP30 cells. 293T cells were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FCS. Madin-Darby canine kidney (MDCK) cells were kept in MEM containing 5% newborn calf serum (NCS). A549 cells were maintained in Kaighn's Modification of Ham's F-12 (F-12K) medium with 10% FCS. All cells were maintained at 37.degree. C. with 5% CO.sub.2.

[0423] Viruses.

[0424] The Ebola.DELTA.VP30 expressing GFP (Ebola.DELTA.VP30-GFP) and Influenzavirus A/WSN/33 (H1N1) were generated, propagated, and titrated as previously described (Halfmann et al., 2008; Neumann et al., 1999). Vesicular stomatitis virus (VSV) strain Indiana, vaccinia virus, and adenovirus Ad-5 were propagated and titrated in Vero cells, CV-1 cells, and A549 cells, respectively.

[0425] High Throughput Screening Assay.

[0426] For compound screening, VeroVP30 cells were seeded in 384-well culture plates. After about 2 hours of incubation at 37.degree. C., 5% CO.sub.2, compounds dissolved in DMSO were added. The cells were then incubated at 37.degree. C. for another approximate 2 hours, before being inoculated with Ebola.DELTA.VP30-GFP virus. All plates included wells to which DMSO was added without any compound for GFP-positive (virus inoculated) and negative (no virus inoculated) controls for the z'-factor calculation. GFP intensities were measured by use of a Safire II plate reader (Tecan Group Ltd., Mannedorf, Switzerland). Cell viabilities were determined by using a CellTiter-Glo.TM. Luminescent Cell Viability Assay (Promega, Madison, Wis., USA) and compared to GFP-positive controls, which were cells treated with DMSO only and inoculated with virus. The high throughput screen was carried out at the Keck-UWCCC Small Molecule Screening Facility (Madison, Wis.).

[0427] Virus Binding and Entry Assay.

[0428] Recombinant VSV viruses, VSV.DELTA.G*-Ebolavirus GP and VSV.DELTA.G*-VSV G, were generated as previously described (Ito et al., 1999; Takada et al., 1997). To determine whether the compounds inhibit virus binding/entry, Vero cells in 12-well plates were treated with 500 .mu.L of 2% FCS-MEM containing 10 .mu.M compounds for 2 hours prior to infection with the recombinant viruses. Since the recombinant virus possesses the GFP reporter gene instead of the VSV G gene, cells expressing GFP after virus inoculation indicate that the virus bound, entered, and replicated the protein in the those cells. Therefore, to determine the efficiency of virus binding/entry mediated by Ebolavirus GP, GFP-positive cells were counted under a fluorescence microscope 16 to 20 hours after virus inoculation and the numbers compared between the two recombinant VSV viruses.

[0429] Virus Minigenome Replication Assay.

[0430] A plasmid-based minireplicon assay was performed as described by Watanabe et al. (Watanabe et al., 2007). To determine whether the compounds inhibit protein expression from the Ebolavirus minigenome, 293T cells were transfected with plasmids for the expression of Ebolavirus nucleoprotein (NP), L, VP35, VP30, Ebolavirus minigenome encoding firefly luciferase, and T7 polymerase. Compounds were added into the media at a final concentration of 10 .mu.M at 6.5 hours post-transfection. Three days post-transfection, cells were disrupted and mixed with Steady Glo (Promega), and luciferase activities were detected by using Glomax (Promega). A reduction in luciferase activity indicates either inhibition of Ebolavirus RNA-dependent RNA polymerase activity or T7 polymerase activity, which is required for Ebola minigenome expression.

Results

[0431] Anti-Ebolavirus High Throughput Compound Screening.

[0432] To identify anti-Ebolavirus compounds, Known Bioactive Library 01, which consists of three commercially available collections totaling 4,160 compounds, was screened with the Ebola.DELTA.VP30-VeroVP30 system. The z'-factor, a measure of assay quality, was consistently over 0.5 and averaged 0.66 (range; 0.50-0.76), indicating that the Ebola.DELTA.VP30-VeroVP30 system was suitable for the HTS assay. Nineteen compounds were identified as anti-Ebolavirus candidates. Six of these were gedunin-like limonoids that shared structural similarities; these six compounds were focused on for further analysis.

[0433] Anti-Ebolavirus Activities of Gedunin and Gedunin-Derivatives.

[0434] Known Bioactive Library 01 contains 41 gedunin-like limonoids. To assess whether all of these compounds show anti-Ebolavirus activity, 39 accessible compounds were re-screened. For this secondary screening, 1.5.times.10.sup.4 cells/30 .mu.L/well were seeded in a 384-well plate, compounds were added at a concentration of 10 .mu.M, and 30 .mu.L of the Ebola.DELTA.VP30-GFP virus was added at an MOI of 0.1, so that the final concentration of the compounds was 5 .mu.M. Fourteen of the compounds reduced the GFP intensity by more than 75%, while cell viabilities were maintained at more than 70%, relative to the positive control (cells that received DMSO and inoculated the virus) (Table 1). The other 25 gedunin-like limonoids tested reduced the GFP intensity by less than 45% (range; 45%-negative 49%) or cell viabilities by more than 95%.

TABLE-US-00001 TABLE 1 % GFP % cell Compound inhibition viability Epoxygedunin* 103 71 Gedunin* 102 77 1,3-Dideacetyl-7-deacetoxy-7-oxokhivorin* 100 78 Dihydrogedunin 96 79 7-Deacetoxy-3-deacetyl-7-oxokhivorin* 92 80 3beta-Acetoxydeoxodihydrogedunin 90 80 Tridesacetoxykhivorin 89 75 3alpha-Hydroxydeoxodihydrogedunin* 85 94 1,3-Dideacetylkhivorin 82 78 Deacetoxy-7-oxogedunin 80 79 Gedunol 79 82 3beta-Hydroxydeoxodihydrogedunin 75 79 1,2alpha-Epoxydeacetoxydihydrogedunin* 75 77 3beta-Hydroxydeoxydesacetoxy-7-oxogedunin 75 81 Heudelottin C 104 5 Deacetylgedunin 45 86 Deacetoxy-7-oxisogedunin 41 84 1,7-Dideacetoxy-1,7-dioxokhivorin 39 88 Isogedunin 35 86 6-Acetoxyangolensic acid methyl ester 32 88 Tridesacetoxykhivorin 28 94 7-Deacetoxy-7-oxokhivorin 26 90 1-Deacetoxy-1-oxo-3,7-dideacetylkhivorin 19 94 6-Hydroxyangolensic acid methyl ester 15 94 1,7-Dideacetoxy-1,7-dioxo-3-deacetylkhivorin 14 87 7-Deacetylkhivorin 13 91 3-Deacetylkhivorin 6 102 Utilin 4 75 7-Epikhivorin 4 79 Angolensic acid, methyl ester 4 85 7-Desacetoxy-6,7-dehydrogedunin 2 70 Khivorin 0 82 Entandrophragmin -12 83 Andirobin -12 91 Prieurianin -17 89 2,3-Dihydroisogedunin -17 91 11alpha-Acetoxykhivorin -47 93 Heudelottin E -49 93 7-Deacetyldihydrogedunin -30 85 *These are the compounds selected for further analyses.

[0435] Comparison of compounds with and without anti-Ebolavirus activity indicated that those compounds with activity against Ebolavirus had a core structure having four benzene rings and a furan ring. In addition, the 1-Keto group of ring A of these compounds may have reduced virus infectivity. For further analysis, five gedunin-like limonoids were selected (FIG. 12; gedunin, epoxygedunin, 1,3-Dideacetly-7-Deacetoxy-7-Oxokhivorin, 7-Deacetoxy-3-deacetyl-7-Oxokhivorin, and 1,2alpha-Epoxy-7-Deacetoxy-7-Oxo-Deoxyhydrogedunin).

[0436] To confirm the anti-Ebolvirus activities of these 5 compounds, the growth kinetics of Ebola.DELTA.VP30-GFP were assessed in their presence. The compounds (10 .mu.M) were added to Vero VP30 cell culture medium 2 hours prior to infection (MOI=0.001) and the medium was then harvested 24, 48, and 72 hours post-infection. As shown in FIG. 13, all five gedunin-like compounds inhibited the growth of Ebola.DELTA.VP30-GFP. Gedunin and epoxygedunin completely inhibited Ebola.DELTA.VP30-GFP growth, while the other three compounds reduced virus growth by at least 1 log.sub.10 titer (85% reduction) at 72 hours post-infection. These data confirm the anti-Ebolavirus activity of these compounds.

[0437] To calculate the anti-Ebolavirus efficacies of the compounds, their 50% inhibitory concentrations (IC.sub.50) were determined by measuring GFP intensity following virus infection at an MOI of 0.1. The compounds showed significant activity with IC.sub.50 values of 0.56 .mu.M or lower, with the exception of 1,2alpha-Epoxy-7-Deacetoxy-7-Oxo-Deoxyhydrogedunin (Table 2), whose IC.sub.50 was 7.12 .mu.M. The 50% cytotoxic concentrations (CC.sub.50) of the compounds were greater than 10 .mu.M, indicating low toxicity to cell culture.

TABLE-US-00002 TABLE 2 IC.sub.50s and CC.sub.50s of Gedunin and Gedunin derivatives compound IC.sub.50 (.mu.M) CC.sub.50 (.mu.M) Gedunin 0.33 >10 Epoxygedunin <0.15 >10 1,3-Dideacetly-7-Deacetoxy-7-Oxokhivorin <0.15 >10 7-Deacetoxy-3-deacetyl-7-Oxokhivorin 0.56 >10 1,2alpha-Epoxy-7-Deacetoxy-7-Oxo- 7.12 >10 Deoxyhydrogedunin

[0438] Virus-Specific Inhibition of Compounds.

[0439] Gedunin and some gedunin-derivatives have antimalarial (MacKinnon et al., 1997), anti-HIV (http://home.ncifcrf.gov/mtdp/Catalog/compounds/309912.html), anti-insect (Nathan et al., 2005), and anti-cancer (Uddin et al., 2007) activities. Therefore, it was determined whether these five compounds inhibited other viruses, namely vaccinia virus, adenovirus, VSV, and influenza virus. Experiments were carried out with 10 .mu.M compounds and infections at an MOI of 0.001 (vaccinia virus, adenovirus) or 0.00001 (VSV and influenza virus). As shown in FIG. 13, none of the compounds significantly inhibited any of the viruses, although gedunin was slightly inhibitory to adenovirus, indicating that the anti-virus activities of these compounds are not universal.

[0440] Inhibition of Ebola GP-Dependent Virus Entry.

[0441] The first step of Ebolavirus infection is virus binding and entry into the host cell via its surface glycoprotein (GP). To examine whether gedunin and gedunin-like compounds inhibit virus entry, we adapted a VSV pseudotype system (Ito et al., 2001; Takada et al., 1997). The pseudotype viruses, VSV.DELTA.G*-Ebolavirus GP and VSV.DELTA.G*-VSV G, possess Ebolavirus GP and VSV G on their surfaces, respectively. Initiation of infection relies upon those surface glycoproteins. In addition, these recombinant viruses possess a GFP reporter gene in place of the VSV G gene, such that infected cells can be distinguished by GFP expression.

[0442] Compounds were added 2 hours prior to the pseudotype virus infections, and virus infectivy was determined by counting the number of GFP-positive cells after an overnight incubation at 37.degree. C. All five compounds appreciably reduced the infectivity of VSV.DELTA.G*-Ebolavirus GP but not that of VSV.DELTA.G*-VSV G (FIG. 14), indicating that these compounds inhibit Ebolavirus GP-dependent virus entry. Interestingly, 2alpha-Epoxy-7-Deacetoxy-7-Oxo-Deoxyhydrogedunin, which inhibited 75% of GFP expression from Ebola.DELTA.VP30-GFP virus (Table 2) and had an IC.sub.50 of 7.12 .mu.M (Table 2), allowed 6.5.+-.1.6% of VSV.DELTA.G*-Ebolavirus GP infection, whereas the other four compounds, which inhibited more than 90% of GFP expression of Ebola.DELTA.VP30-GFP virus infection (Table 2) and had IC.sub.50s more than 10 times lower than that of 2alpha-Epoxy-7-Deacetoxy-7-Oxo-Deoxyhydrogedunin, allowed less than 3% of VSV.DELTA.G*-Ebolavirus GP infection. These data suggest that the level of Ebolavirus inhibition of these compounds is associated with their ability to inhibit Ebolavirus GP-dependent virus entry.

[0443] Inhibition of Protein Expression from the Ebolavirus Minireplicon.

[0444] The next steps in Ebolavirus replication are virus genome replication and virus protein expression. To examine whether the tested compounds inhibit Ebolavirus genome replication and protein expression, an Ebolavirus minigenome replication assay was performed. The compounds were added to cell culture media 6.5 hours post-transfection to avoid affecting transfection efficacies. As shown in FIG. 15, gedunin and epoxygedunin significantly reduced firefly luciferase reporter protein expression from the Ebolavirus minireplicon. The luciferase activities in cells treated with these two compounds were 1.3%.+-.0.2% and 1.0%.+-.0.1% of those treated with DMSO, respectively. The other three compounds did not reduce the luciferase activities. Since gedunin and epoxygedunin have a 7-acetate group on their ring B, but the other three compounds do not, this residue may contribute to the inhibition of Ebolavirus genome replication and/or protein expression.

[0445] Hsp90 Inhibitors Reduce Protein Expression from the Ebolavirus Minireplicon.

[0446] The inhibitor activities of gedunin and some of its derivatives have been tied to the heat shock protein Hsp90 (Hieronymus et al., 2006), suggesting that their Ebolavirus inhibitory mechanisms may involve inhibition of Hsp90 or degradation of its substrate proteins. Therefore, it was determined whether Hsp90 inhibitors have anti-Ebolavirus activity. Four Hsp90 inhibitors, geldanamycin (GM), 17-AAG (17-Allylamino-17-demethoxygeldanamycin), CCT 018159 (4-[4-(2,3-Dihydro-1,4-benzodioxin-6-yl)-5-methyl-1H-pyrazol-3-yl]-6-ethy- l-1,3-benzenediol), and AEG 3482 (6-Phenylimidazo[2,1-b]-1,3,4-thiadiazole-2-sulfonamide) were assessed for their anti-Ebolavirus activities by use of an Ebolavirus growth assay, a VSV pseudotype virus assay, and a minireplicon assay.

[0447] CCT and 17AAG reduced Ebola.DELTA.VP30 virus growth (97% and 92% reduction, respectively, at 72 hours post-infection), but GM and AEG did not (30% and 5% reduction, respectively, 72 hours post-infection) (FIG. 16A). Although all five of the gedunin-like compounds significantly reduced virus infection mediated by Ebolavirus GP, only CCT 018159 of the Hsp90 inhibitors slightly reduced the Ebolavirus GP-mediated virus infection (FIGS. 16B and 16C). Infectivities of VSV.DELTA.G*-Ebolavirus GP, which were standardized by the infectivities of VSV.DELTA.G*-VSV G, were 137% (GM), 110% (17-AAG), 71% (CCT 018159) and 135% (AEG 3482). All four Hsp90 inhibitors reduced reporter protein expression from the Ebolavirus minigenome [luciferase activities were 7.9%.+-.1.5% (17-AAG), 8.8%.+-.2.6% (CCT 018159), 24.7%.+-.7.7% (GM), and 34.0%.+-.8.0% (AEG 3482)](FIG. 16D). These data demonstrate that Hsp90 inhibitors are potential anti-Ebolavirus agents and that their inhibitory mechanisms likely differ from those of gedunin and its derivatives.

Discussion

[0448] A high throughput molecular screen for anti-Ebolavirus agents identified gedunin and its derivatives as anti-Ebolavirus candidates. Further analysis demonstrated that these compounds inhibit Ebolavirus via Ebolavirus GP-dependent virus binding/entry and that some of them also reduce Ebolavirus genome replication and/or protein expression. Gedunin-like limonoids are found in extracts of plants from the Meliaceae (Mahogany) family and have been used in traditional medicine in tropical America and in West and East Africa (Bray et al., 1990), suggesting that there is potential for their use in humans, if in vivo experiments confirm their anti-Ebolavirus activities.

[0449] In this study, the specificity of anti-Ebolavirus compounds was assessed by testing their inhibitory activities against influenzavirus, VSV, vaccinina virus, and adenovirus because these compounds are reported to have antimalarial (MacKinnon et al., 1997), anti-HIV (http://home.ncifcrf.gov/mtdp/Catalog/compounds/309912.html), anti-insect (Nathan et al., 2005), and anti-cancer (Uddin et al., 2007) activities. However, none of the compounds tested exhibited significant inhibition of these viruses, although gedunin did slightly delay the propagation of vaccinia virus, adenovirus and VSV. The structural features of these compounds may be a determinant of specificity since only 14 of the 41 gedunin-like limonoids that were screened demonstrated inhibitory activity to Ebolavirus and since 7-deacetoxy-7-hydroxygedunin and 7-deacetoxy-7-oxogeduin had been identified as anti-HIV compounds but the other gedunin-derivatives had not (http://www.stjuderesearch.org/guy/data/parasite_bioactives_screen/MAL_3D- 7/Results/87.html). These data suggest that gedunin-like limonoids have potential as general antivirals and further screening of these compounds using other microbial assays may be of value.

[0450] The mechanisms by which gedunin and its derivatives inhibit Ebolavirus remain unknown; it is not clear whether they interact with host cell components or with Ebolavirus proteins and/or genomes. Since it was previously reported that gedunin and its derivatives express anti-cancer activities via degradation of Hsp90 and/or its substrates (Hieronymus et al., 2006) and DNA and RNA virus propagation can be delayed by Hsp90 inhibitors (Basha et al., 2005; Burch & Weller, 2005; Chase et al., 2008; Connor et al., 2007; Li et al., 2004; Ujino et al., 2009), it seemed possible that Hsp90 inhibitory activities may contribute to the anti-Ebolavirus activities of gedunin and its derivatives. Therefore, the anti-Ebolavirus activities of Hsp90 inhibitors was examined.

[0451] Although the four Hsp90 inhibitors tested did not inhibit Ebolavirus GP-dependent virus binding/entry to the same extent as the gedunin-like compounds, they reduced protein expression from the Ebolavirus minireplicon and two of the four Hsp90 inhibitors also delayed Ebola.DELTA.VP30-GFP replication. Since structurally different compounds have been found with Hsp90 inhibitors to limit reporter protein expression from the Ebolavirus minireplicon, it has been suggested that this inhibition may be due not to structural binding to Ebolavirus directly, but to Hsp90 inhibitory activities. However, mechanisms other than Hsp90 inhibition should be considered since deacetylegedunin, which shows anti-cancer activity via degradation of Hsp90 substrates (Hieronymus et al., 2006) was one of the 41 gedunin-like limonoids tested, yet it did not show any anti-Ebolavirus activity.

[0452] CCT 018159 displayed about a 30% reduction in Ebolavirus GP-dependent virus infection, unlike GM, 17-AAG, and AEG 3482. It has been reported that CCT 018159 binds to the ATP site located in the N-terminal domain of Hsp90; however, GM and 17-AAG also bind at this location via the same main amino acids (Cheung et al., 2005; Stebbins et al., 1997). Therefore, why only CCT 018159 showed inhibitory activity to Ebolavirus binding/entry is unclear. The data suggest that blockage of virus binding/entry mediated by Ebolavirus GP may not rely upon Hsp90 and/or its substrate/signaling.

[0453] Inhibitors of S-adenosylhomocysteine hydrolase (SAH) have also shown anti-Ebolavirus activity in vitro and in vivo (Huggins et al., 1999). Their IC.sub.50 values range from 2 to 64 .mu.M, which is higher than gedunin and its derivatives in this study. Although it is not strictly valid to directly compare these values, since they were determined using different assays, IC.sub.50 values are not assay-dependent or Ebolavirus strain-dependent (Huggins et al., 1999). Therefore, the anti-Ebolavirus efficacies of gedunin and its derivatives are at least equal to those of SAH inhibitors.

[0454] Ebolavirus outbreaks have occurred almost every year in the 21.sup.st century in Africa, infecting and killing numerous individuals. Moreover, many people and pigs were infected with Ebolavirus Reston in the Philippines in 2008-2009. These reports reflect that Ebolavirus infection is an ongoing threat and that therapeutic and prophylactic options are desperately needed. Gedunin-like limonoids are found in traditional medicine in tropical and subtropical regions, where they have been used to treat humans. The compounds identified in the present screen thus show promise as anti-Ebolavirus agents, and in vivo confirmation of their anti-Ebolavirus activities is warranted.

Summary

[0455] A library of compounds (Known Bioactive Library (KB01)) at the Keck-UWCCC Small Molecule Screening Facility, University of Wisconsin-Madison, Madison, Wis.) was screened to determine whether any of the compounds interfered with Ebolavirus replication and infection. The compounds were screened using biologically contained Ebolavirus expressing GFP protein (Ebola.DELTA.VP30-GFP virus). In particular, gedunin and gedunin-like compounds were screened. Those compounds are found in extracts of plants from the Meliaceae (Mahogany) family and have been used in traditional medicine for the treatment of fevers in tropical American and in West and East Africa. They are known for their antimalarial, anti-HIV, anti-cancer, and anti-insect activities. Gedunin is an inhibitor of Hsp90.

[0456] Through the use of Ebola.DELTA.VP30-GFP virus and a follow-up screen, 15 gedunin and gedunin-like compounds were identified that reduced GFP expression by at least 75%.

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[0564] All publications, patents and patent applications are incorporated herein by reference. While in the foregoing specification, this invention has been described in relation to certain preferred embodiments thereof, and many details have been set forth for purposes of illustration, it will be apparent to those skilled in the art that the invention is susceptible to additional embodiments and that certain of the details herein may be varied considerably without departing from the basic principles of the invention.

Sequence CWU 1

1

33118875DNASudan ebolavirus 1cggacacaca aaaagaaaga aaagtttttt atactttttg tgtgcgaata actatgagga 60agattaatca ttttcctcaa actcaaacta atattaacat tgagattgat ctcatcattt 120accaattgga gacaatttaa ctagtcaatc ccccatttgg gggcattcct aaagtgttgc 180aaaggtatgt gggtcgtatt gctttgcctt ttcctaacct ggctcctcct acaattctaa 240cctgcttgat aagtgtgatt acctgagtaa tagactaatt tcgtcctggt aattagcatt 300ttctagtaaa accaatacta tctcaagtcc taagagaagg tgagaagagg gtcccgaggt 360atccctccag tccacaaaat ctagctaatt ttagctgagt ggactgatta ctctcatcac 420acgctaacta ctaagggttt acctgagagc ctacaacatg gataaacggg tgagaggttc 480atgggccctg ggaggacaat ctgaagttga tcttgactac cacaaaatat taacagccgg 540gctttcggtc caacaaggga ttgtgcgaca aagagtcatc ccggtatatg ttgtgagtga 600tcttgagggt atttgtcaac atatcattca ggcctttgaa gcaggcgtag atttccaaga 660taatgctgac agcttccttt tacttttatg tttacatcat gcttaccaag gagatcatag 720gctcttcctc aaaagtgatg cagttcaata cttagagggc catggtttca ggtttgaggt 780ccgagaaaag gagaatgtgc accgtctgga tgaattgttg cccaatgtca ccggtggaaa 840aaatcttagg agaacattgg ctgcaatgcc tgaagaggag acaacagaag ctaatgctgg 900tcagttttta tcctttgcca gtttgtttct acccaaactt gtcgttgggg agaaagcgtg 960tctggaaaaa gtacaaaggc agattcaggt ccatgcagaa caagggctca ttcaatatcc 1020aacttcctgg caatcagttg gacacatgat ggtgatcttc cgtttgatga gaacaaactt 1080tttaatcaag ttcctactaa tacatcaggg gatgcacatg gtcgcaggcc atgatgcgaa 1140tgacacagta atatctaatt ctgttgccca agcaaggttc tctggtcttc tgattgtaaa 1200gactgttctg gaccacatcc tacaaaaaac agatcttgga gtacgacttc atccactggc 1260caggacagca aaagtcaaga atgaggtcag ttcattcaag gcagctcttg gctcacttgc 1320caagcatgga gaatatgctc catttgcacg tctcctcaat ctttctggag tcaacaactt 1380ggaacatggg ctttatccac aactctcagc cattgctttg ggtgttgcaa ctgcccacgg 1440gagcacgctg gctggtgtta atgtagggga gcaatatcag caactgcgtg aggctgctac 1500tgaagctgaa aagcaactcc aacaatatgc tgaaacacgt gagttggaca accttgggct 1560tgatgaacag gaaaagaaga ttctcatgag cttccaccag aagaagaatg agatcagctt 1620ccagcaaact aacgcaatgg taaccctgag gaaagagcgg ctggccaaac tgaccgaagc 1680catcacgact gcatcaaaga tcaaggttgg agatcgttat cctgatgaca atgatattcc 1740atttcccggg ccgatctatg atgaaaccca ccccaaccct tctgatgata atcctgatga 1800ttcacgtgat acaactatcc caggtggtgt tgttgacccg tatgatgatg agagtaataa 1860ttatcctgac tacgaggatt cggctgaagg caccacagga gatcttgatc tcttcaattt 1920ggacgacgac gatgacgaca gccaaccagg accaccagac agggggcaga gcaaggaaag 1980agcggctcgg acacatggcc tccaagatcc gaccttggac ggagcgaaaa aggtgccgga 2040gttgacccca ggttcccacc aaccaggcaa cctccacatc accaagccgg gttcaaacac 2100caaccaacca caaggcaata tgtcatctac tctccagagt atgaccccta tacaggaaga 2160atcagagccc gatgatcaga aagatgatga tgacgagagt ctcacatccc ttgactctga 2220aggtgacgaa gatgttgaga gcgtatcagg ggagaacaac ccaactgtag ctccaccagc 2280accagtctac aaagatactg gagtagacac taatcagcaa aatggaccaa gcaatgctgt 2340agatggtcaa ggttctgaaa gtgaagctct cccaatcaac cccgaaaagg gatctgcact 2400ggaagaaaca tattatcatc tcctaaaaac acagggtcca tttgaggcaa tcaattatta 2460tcacctaatg agtgatgagc ccattgcttt tagcactgaa agtggcaagg aatatatctt 2520cccagattct cttgaagaag cctacccgcc ttggttgagt gagaaggagg ccttagagaa 2580agaaaatcgt tatctggtca ttgatggcca gcaattcctc tggccagtaa tgagcctaca 2640ggacaagttc cttgctgttc ttcaacatga ctgaggaccc atgattagta gattttgttt 2700attctgagct tgattataat tgttttgata attcaagtat gagcaaccaa cccgaaatat 2760aaaccctatt ttagttatga ggaaattaaa taaataatct gtaagttgta ggactatgaa 2820gagctgcttg tgtcaattta tcacgggcta atacccatac cgcaagaata attatttagt 2880aattttgatc agcttatgat atgtaccaat aggaaaacat tatagcatta aaacataaag 2940tatccttcga tgagcttagg aggataatat cctgatgaat tctatagaac ttaggattaa 3000gaaaaaattc atgatgaaga ttaaaacctt catcatcctt taaaaagaga gctattcttt 3060atctgaatgt ccttattaat gtctaagagc tattattttg taccctctta gcctagacac 3120tgcccagcat ataagccatg cagcaggata ggacttatag acatcatgga cccgaagtgt 3180ctggctggtt ttctgagcaa ttaatgaccg gcaaaatacc gctaacagag gtgtttgttg 3240atgttgaaaa caaaccaagt cctgccccga taaccattat tagtaagaat cccaagacaa 3300cacgtaaaag tgataagcaa gtccaaacag atgatgccag tagcttattg acagaagaag 3360tcaaggctgc cataaattcg gtgatatcag ctgtgcgtcg gcaaaccaat gctattgaat 3420cactagaagg tcgagtaaca actcttgagg ccagcttaaa acccgttcaa gacatggcaa 3480agaccatatc atccctgaat cgcagctgtg ccgaaatggt tgcaaaatac gacctactgg 3540tgatgaccac tgggcgagca actgccactg ctgcagcaac agaagcatat tggaatgaac 3600atggacaagc acctccaggc ccatcattgt acgaggatga tgctattaag gctaaattga 3660aagatccgaa cgggaaggtt ccagaaagtg tcaaacaggc ctacataaat ctagatagca 3720caagtgccct caatgaggaa aatttcgggc gaccttacat ttcagcaaaa gatctcaagg 3780aaatcatcta tgaccatctc ccaggatttg ggacagcttt tcatcagttg gtgcaggtta 3840tctgcaaaat tggtaaggat aataatatcc tagacataat tcatgcagaa ttccaagcaa 3900gcttggctga gggagactcc ccccagtgtg cattaatcca gataacaaaa cggatccctg 3960ctttccaaga tgcctctcct ccaattgtgc atatcaagtc tcgaggagat atacccaaag 4020cctgtcagaa aagcctccgg ccggtcccac cgtcaccaaa gatcgataga ggttgggtct 4080gtatttttca attccaagac gggaaggccc ttgggctaaa aatatgatac agaagcaagg 4140taagctcatt ttgcgatggc caaatgatac ttatgactgt ttaaaatcaa gttagactaa 4200tagtctattg tgtcataagc ttataagtca gttttaaatt tcccctctat cctaatcaat 4260tgataatgct gtcaataggg aaattcccct gtattgtaat aagacctcat taacatattt 4320cccctgctta gtactatgca gaaacccccg agcaaattaa aattgatgaa gattaagaaa 4380aagagggatt ttctcaggaa aaatcttttt tcttaccttc atctcattta aacaaattta 4440ggactcagga aaaatgagaa gggtcactgt gccgactgca ccacctgcct atgctgacat 4500tggctatcct atgagcatgc ttcccatcaa gtcaagcagg gctgtgagtg gaattcaaca 4560gaaacaagag gtccttcctg gaatggatac accatcaaat tctatgagac ctgttgctga 4620tgataacatt gatcatacaa gtcatacccc gaacggagtg gcctcagcat tcatcttgga 4680ggcaactgtc aatgtgatct cggggcccaa agtcctcatg aaacaaatcc ctatttggtt 4740gccactcgga attgctgacc aaaaaacgta cagttttgac tcaacaacag cagcaattat 4800gctcgcatct tatacgatca cccattttgg aaaggccaac aaccccctcg ttagagtgaa 4860tcgacttggt cagggaatac cggatcaccc actcagattg ctcaggatgg ggaaccaggc 4920tttccttcaa gagtttgtgc taccaccagt tcaactgccg caatatttca cttttgatct 4980gactgcactc aaactagtga cacagcctct ccctgctgca acatggacag atgagactcc 5040gagcaacctt tcaggagccc ttcgtcccgg gctttcattt cacccaaagc tgagacccgt 5100tctacttcca ggcaagacgg gaaagaaagg gcatgtttct gatctgactg ccccagacaa 5160aattcagaca attgtgaacc tgatgcaaga tttcaaaatc gtgccaattg atccagctaa 5220gagtatcatt gggatcgagg ttccagaatt gctggtccac aagctcactg ggaagaaaat 5280gagtcagaag aatggacagc ctataattcc tgtcttactc ccaaaataca ttgggctaga 5340tccaatctca cctggagacc tgactatggt cataacacca gattatgatg attgtcattc 5400acctgccagt tgctcttatc tcagtgaaaa gtgattctca caaagtgaga gaaacacctc 5460cagtaaagaa atcaaatctt atctatagca actcaatcga cttttaggaa gctagcagtc 5520catatactat gggacaactc aaccctcttg ttaaaatgta ctaatcgggt caaggaactc 5580tcactgatca agcctgaatc caagatagaa ccagcccaaa gggcctcccc agagtctctt 5640acaagcttag ccaatcaatt aacatgcata agcgatccat acttcgccca atcagtgtcc 5700gatgttcacc ccttcaagcc tccttcctag caaattgacc tagctgtacc aagagattcc 5760ctcagcctcc ttctcaaata acctgatcct cgagggttac accttcacca ctctatgctc 5820atttcaccca aacataaaat gaaatgtctt aacatgattg caccattaag aaaaacaaat 5880ctgatgaaga ttaagcctga tgaaggccca accttcatct ttttaccata atcttgttct 5940cagtaccatt tgataagggt acacttgcca atacgccccc atcctaaggg tctcgcaatg 6000gggggtctta gcctactcca attgcccagg gacaaatttc ggaaaagctc tttctttgtt 6060tgggtcatca tcttattcca aaaggccttt tccatgcctt tgggtgttgt gactaacagc 6120actttagaag taacagagat tgaccagcta gtctgcaagg atcatcttgc atctactgac 6180cagctgaaat cagttggtct caacctcgag gggagcggag tatctactga tatcccatct 6240gcaacaaagc gttggggctt cagatctggt gttcctccca aggtggtcag ctatgaagcg 6300ggagaatggg ctgaaaattg ctacaatctt gaaataaaga agccggacgg gagcgaatgc 6360ttacccccac cgccagatgg tgtcagaggc tttccaaggt gccgctatgt tcacaaagcc 6420caaggaaccg ggccctgccc aggtgactac gcctttcaca aggatggagc tttcttcctc 6480tatgacaggc tggcttcaac tgtaatttac agaggagtca attttgctga gggggtaatt 6540gcattcttga tattggctaa accaaaagaa acgttccttc agtcaccccc cattcgagag 6600gcagtaaact acactgaaaa tacatcaagt tattatgcca catcctactt ggagtatgaa 6660atcgaaaatt ttggtgctca acactccacg acccttttca aaattgacaa taatactttt 6720gttcgtctgg acaggcccca cacgcctcag ttccttttcc agctgaatga taccattcac 6780cttcaccaac agttgagtaa tacaactggg agactaattt ggacactaga tgctaatatc 6840aatgctgata ttggtgaatg ggctttttgg gaaaataaaa aaatctctcc gaacaactac 6900gtggagaaga gctgtctttc gaagctttat cgctcaacga gacagaagac gatgatgcgg 6960catcgtcgag aattacaaag ggaagaatct ccgaccgggc caccaggaag tattcggacc 7020tggttccaaa gaattcccct gggatggttc cattgcacat accagaaggg gaaacaacat 7080tgccgtctca gaattcgaca gaaggtcgaa gagtaggtgt gaacactcag gagaccatta 7140cagagacagc tgcaacaatt ataggcacta acggcaacca tatgcagatc tccaccatcg 7200ggataagacc gagctccagc caaatcccga gttcctcacc gaccacggca ccaagccctg 7260aggctcagac ccccacaacc cacacatcag gtccatcagt gatggccacc gaggaaccaa 7320caacaccacc gggaagctcc cccggcccaa caacagaagc acccactctc accaccccag 7380aaaatataac aacagcggtt aaaactgtcc tgccacagga gtccacaagc aacggtctaa 7440taacttcaac agtaacaggg attcttggga gtcttgggct tcgaaaacgc agcagaagac 7500aaactaacac caaagccacg ggtaagtgca atcccaactt acactactgg actgcacaag 7560aacaacataa tgctgctggg attgcctgga tcccgtactt tggaccgggt gcggaaggca 7620tatacactga aggcctgatg cataaccaaa atgccttagt ctgtggactt aggcaacttg 7680caaatgaaac aactcaagct ctgcagcttt tcttaagagc cacaacggag ctgcggacat 7740ataccatact caataggaag gccatagatt tccttctgcg acgatggggc gggacatgca 7800ggatcctggg accagattgt tgcattgagc cacatgattg gacaaaaaac atcactgata 7860aaatcaacca aatcatccat gatttcatcg acaacccctt acctaatcag gataatgatg 7920ataattggtg gacgggctgg agacagtgga tccctgcagg aataggcatt actggaatta 7980ttattgcaat tattgctctt ctttgcgttt gcaagctgct ttgctgaata tcaatttgaa 8040tcatcaattt aagcttgata catttctagc attttaaatt ataaaccgat actgatactt 8100gaaaatcagg ctaatgccaa gttctgtgca aaacttgaaa gtaggtttac aaaaatcctt 8160tggactggaa tgctttgata ctctttctca atactatata agttccttcc caagaataat 8220attgatgaag attaagaaaa agtgacattg tgcccacttt tgtaatcttc atccacctac 8280acattcatat tcaggaatct ttgaattaac cctcacactt gcttaggaaa gagcctatcc 8340tctacaagaa tcccgaggcg gcaattcagt taatttcata tcaagataac atccatttcc 8400aagaccacag ataactatat tattaatctt taccacaaat atggagaggg gtcgtgagcg 8460cgggagatca aggaattcac gtgccgacca gcaaaattca acaggtcctc aatttaggac 8520aagatccatt tcccgggata agacaacaac agactaccgt agtagtcgaa gtacttcgca 8580agttagagtc cctacggttt tccataagaa aggtactggg acccttactg tccctccagc 8640acctaaggat gtttgtccta ctctcagaaa aggatttcta tgtgatagta atttctgtaa 8700aaaggaccat caacttgaaa gcctaaccga ccgggagctc ctacttctta tagcacggaa 8760gacctgtgga tcaactgatt catcgcttaa tatagctgct cctaaagacc taagactagc 8820aaatcctacg gctgatgact tcaagcaaga cggcagtcca aaattaaccc taaaattact 8880agtcgagact gctgagtttt gggccaatca gaatattaat gaagtagatg atgcaaaact 8940ccgtgctctc ttgacgttga gtgctgtctt agtgcggaaa ttctctaagt cacagcttag 9000tcaattatgt gagagtcatc ttaggaggga aaacttagga caagaccaag ctgaatcagt 9060tctcgaggtt tatcaacgtt tacatagtga caaaggaggt gcttttgagg cagcactatg 9120gcaacagtgg gatagacaat cattaactat gtttatatct gctttcctcc atgtagcatt 9180gcaactttcc tgtgagagct ccactgtagt gatatcaggc ctacgcttac ttgccccccc 9240aagcgttaat gaagggctcc ctcctgcacc aggggaatat acttggtcag aagatagtac 9300aacttagcct gtagggagga caagtaaaac aagatgccct tatcctctat agatggtatt 9360tttagagagg gggacaggat aggaataaag ataatgacta aagccaatat aaagatacga 9420acacaagtag aaattaaaat agaaatcaaa acaatctccc cttattcaat atgaaatata 9480atagtgagta tttgtttcat gatgtcaatc atttattgtt aaaaataaac aaagtcagta 9540agagtgttag gatcgttata ttgcaaggat cctccctaga agcgttgaat catctcaagt 9600agcctagaac aagaacagca gagcattaaa ttgaaataga taataaggat attgcttgtt 9660tttaagatag ttttaggaag tttaaaatta agaaaaagaa cccatggaca cactctagca 9720ttgaggatgg ggttcccttg atgatagtat agtcttaggt atagggtagt cctacacgta 9780ctatattata cagtctaaac ttgtaaaatt aaactacaag aacatgatga aaattaatga 9840gaaggttcca agattgactt caatccaaac accttgctct gccaattttc atctccttaa 9900gatatatgat tttgttcctg cgagataagg ttatcaaata gggtgtgtat ctcttttaca 9960tatttgggct cccactaggc tagggtttat agttaaggaa gactcatcac attttttatt 10020gaactagtct actcgcagaa tcctaccggg aatagaaatt agaacatttg tgatactttg 10080actataggaa ataattttca acactacctg agatcaggtt attcttccaa cttattctgc 10140aagtaattgt ttagcatcat aacaacaacg ttataattta agaatcaagt cttgtaacag 10200aaataaagat aacagaaaga acctttatta tacgggtcca ttaattttat aggagaagct 10260ccttttacaa gcctaagatt ccattagaga taaccagaat ggctaaagcc acaggccggt 10320acaacttggt aacaccaaaa cgggagctag agcaaggagt tgtgtttagc gacctatgca 10380acttcctagt gactccaact gtgcaaggat ggaaggttta ctgggctgga cttgagtttg 10440atgtcaacca aaagggtatt accctgttaa atcgtcttaa agtgaatgat tttgctcctg 10500catgggcgat gacccggaac ctcttcccac acttgttcaa aaaccaacag tctgaagtcc 10560aaactcccat ttgggccttg agggtaattc ttgccgccgg gattcttgac caattaatgg 10620atcattccct cattgagccg ctatcagggg ccctgaacct aattgctgat tggttactaa 10680caacatctac taatcacttc aacatgagaa ctcaacgagt aaaggaccaa ctgagcatga 10740ggatgttatc tcttataagg tcaaatatta ttaactttat aaataagctc gagactcttc 10800atgtcgttaa ttacaaggga cttctaagca gtgttgagat aggaacacca agctatgcaa 10860tcatcattac caggactaat atgggttatc ttgtcgaggt tcaggaacca gataaatctg 10920cgatggatat acgacaccct ggtcctgtca aattctcctt actacatgaa tcgacactta 10980aacctgttgc cactcctaaa ccatcaagca ttacttcatt gatcatggag ttcaacagtt 11040ctttggcaat ttaattgccg taataaaaat tgtacgatag ggctaacatt gattccataa 11100tccatcgtag gacagaatca ttttcctgta tgatcttagt ttaatctctc tttatacaat 11160gattaataag gagcctgttt aaaatgttac aaaagtatac tgtttgaacc cctagtatcc 11220ctgtaaatat cctcattcaa ttttttgctt ttacatgtgt agtcacctgt atagcatgac 11280cctagtcatg cctttaatta atacttaatc taacagttaa tataatgtat aactttccat 11340gttcaaagag tagtcaaaac aatgtgagat ccagtttcac tcacagcatc tattcactat 11400ttacagtatg atgagcccaa attaacacgg tagaggtcta gatttattaa tagaacgagg 11460aagattaaga aaaagtccat aatgctgggg aggcaatcct tgccaccata ggactttttc 11520aattcctcta ttttatgatg gctacccaac atacacaata tcctgatgca agattgtctt 11580ccccaattgt cttagaccaa tgtgacctag tgacaagagc atgtggactt tactctgagt 11640attcgctgaa ccctaaacta aagacatgcc gtttaccgaa acatatctat agattaaaat 11700atgacactat tgttttacga tttattagtg atgtccctgt agctacaatc ccaatagact 11760acattgctcc gatgttaata aatgttctgg cagatagtaa aaatgtacca ttggaacctc 11820cctgcttgag tttcttggat gaaatagtca attataccgt gcaggatgca gccttcctta 11880attattacat gaatcagatt aaaacacagg aaggagtaat tacagatcaa ttaaaacaga 11940acattcgtag ggtcattcac aaaaacagat atctatctgc tctattcttc tggcatgatc 12000ttgccatcct cacccgtcga gggagaatga accgaggaaa tgtgcgctcc acttggtttg 12060taacgaatga ggttgttgac attctaggat atggtgatta tatcttctgg aagatcccta 12120ttgctctatt accaatgaac acagctaatg ttccacatgc atcaactgac tggtaccaac 12180ctaatatctt caaggaggct attcaaggac acacacatat tatttcagtc tctacagccg 12240aggtccttat tatgtgtaag gatcttgtca caagtcgttt taataccctt ctgattgctg 12300agttagccag gttggaagat ccagtgtctg ctgattatcc actagtagat aatattcaat 12360ctctgtataa cgcaggagac tacctgttgt ccatattggg atcagagggg tacaaaataa 12420tcaaatatct cgaacctctg tgtttggcta agattcaact atgttcccaa tatacagaac 12480gaaaagggcg gtttttaacc cagatgcatc ttgcagttat tcagacattg cgtgaactcc 12540tccttaatag agggttgaaa aaatcacaat tgtctaaaat ccgcgagttt caccaactgt 12600tgctcagact ccgatctaca ccacaacaat tatgtgaatt attttcaatc caaaaacact 12660ggggccaccc agttctgcat agtgaaaagg ccatccaaaa ggttaaaaat catgcaacag 12720ttctaaaggc attgcggccg attatcatct ttgaaacgta ttgtgtattc aagtatagtg 12780ttgcaaaaca tttctttgat agtcaaggca cttggtacag tgtgatatca gaccgatgtt 12840taacgccggg attgaattcc tacattaggc gaaatcaatt ccctccactt ccaatgatca 12900aagatctttt atgggaattt taccatttgg atcatcctcc attattctcc acgaagatca 12960ttagtgacct cagcattttc attaaagacc gcgcaacagc agttgaacaa acctgttggg 13020atgcagtttt tgagcctaac gttttgggct acagtccacc ttatcgattc aataccaaac 13080gtgtacctga acaattcctg gagcaagagg atttttctat tgagagtgtc ttacaatacg 13140cccaagaact taggtactta ttgccccaga atcgaaattt ttctttttca ttgaaggaaa 13200aagaattaaa tgttggtagg acatttggaa aattgcctta tttaaccagg aatgtccaaa 13260ccctctgcga agcattactt gcagatggtt tggctaaagc ctttccaagc aatatgatgg 13320ttgtcacaga gagggaacaa aaggagagcc tccttcacca agcatcctgg caccatacaa 13380gtgatgattt cggagagcat gccacagttc gtggaagtag ttttgtcaca gacctggaaa 13440aatacaatct ggccttcagg tatgaattca cagctccctt catcaaatat tgcaaccaat 13500gctatggggt tcgcaatgtc tttgattgga tgcacttcct aattccgcaa tgttacatgc 13560atgttagtga ttattataac ccaccacata atgtaacctt agagaatagg gaatatcccc 13620ccgaaggacc aagtgcttat agaggccacc ttggcggtat tgaggggctt caacaaaagt 13680tatggactag tatctcatgt gctcaaatct cattggtaga gatcaagacc gggttcaaat 13740tgcgatcagc agtcatgggg gataatcaat gtattacagt attatcagtc tttccactag 13800aatctagtcc gaatgagcag gagagatgcg cagaagacaa tgcagccaga gtggctgcta 13860gcttggccaa agtcacaagt gcctgtggga tattcctcaa gcctgatgag actttcgtac 13920actcaggctt tatctatttt ggcaaaaagc aatacttgaa cggaattcaa ttacctcaat 13980cactcaagac agcagctagg atggcccctc tctcagatgc aatttttgat gacttgcaag 14040gtacacttgc cagtatagga actgcctttg agcgatcaat ctccgaaact agacatattt 14100taccatgccg tgttgcagct gcctttcata catatttctc tgttcggatc ttacaacatc 14160atcaccttgg tttccataag ggttcagacc ttggacaatt ggcaatcaat aaacctcttg 14220atttcgggac cattgcacta tccttagcag ttcctcaggt attgggtgga ttatccttcc 14280taaatccaga aaagtgcctt tatcgcaact tgggtgatcc tgtaacttca ggcctatttc 14340agttgaagca ttatctgtca atggtgggta tgagtgatat ctttcatgca cttattgcaa 14400aaagcccagg gaattgtagc gcaattgact ttgttctaaa cccaggcggg ttaaatgtcc 14460ctggatcaca ggatttaaca tctttccttc gtcagattgt cagaaggagt atcacacttt 14520cggcaaggaa caagttaatc aacacgttat ttcacgcttc tgcagatctt gaagacgaat 14580tagtatgtaa atggttactt tcttcaacgc ccgtgatgag ccgttttgca gccgatattt 14640tctcacgaac accaagcggg aaaagattac aaatcttggg atacctcgag ggaaccagaa 14700ctttattagc atccaaaatg ataagcaata atgcagagac accaatcttg gagaggctca 14760gaaaaataac acttcaaaga tggaatctat ggtttagtta cctagaccat tgtgacccag 14820ctttaatgga agcaattcaa ccaattaagt gtactgttga tattgctcaa attcttagag 14880aatactcctg ggctcatatc cttgatggta gacagttaat aggggcaaca ctgccatgta 14940tacctgagca gttccaaacc acatggttaa aaccttacga gcaatgtgtg gaatgttcat 15000ccacaaacaa ttctagtcca tatgtatcag ttgcattaaa

aaggaacgtg gttagtgctt 15060ggcctgatgc atctagattg gggtggacga ttggtgatgg gattccctac ataggctcaa 15120gaactgagga caaaataggt cagcccgcta ttaagccgag gtgcccatca gctgcattaa 15180gagaagctat tgaattgacc tctaggttga cctgggtcac tcaaggtagt gcaaacagcg 15240atcagttaat tcgccctttt cttgaggcaa gagtaaactt gagtgtacaa gagattcttc 15300aaatgacccc ctcacattac tccggtaata ttgtgcatcg gtataatgat cagtatagcc 15360ctcactcctt tatggctaac cgcatgagta acacagcaac gcgcttgatg gtatctacca 15420acacactagg agagttttcc ggagggggtc aggctgcacg tgatagcaac attatatttc 15480aaaatgtgat taactttgca gtggccttgt atgacattag gtttcggaac acttgtacat 15540cttctattca atatcacagg gcccatattc acctgacgaa ttgttgtacg agggaagtac 15600cggcccaata cttaacatac acaaccacgc taaatctaga tttgagtaag taccgtaata 15660atgaactgat ttatgattca gatccactaa gaggaggtct caactgcaac ttatcgattg 15720acagtccttt gatgaagggc ccacgtttaa atattattga ggatgactta atacggttgc 15780cacatttatc cggctgggaa ttagcaaaaa cagtcttgca atcaataatc tctgatagta 15840gcaattcatc aacagatccc attagcagcg gtgaaacaag atccttcaca acccacttct 15900taacgtatcc caaaataggg cttctataca gttttggagc cctcataagt ttttatttgg 15960gtaatactat tctatgcacg aaaaagatcg gactcacaga atttctatac tatctccaga 16020atcagatcca caacttatca catagatccc ttcgaatctt caaaccgaca tttagacact 16080caagtgtcat gtccaggttg atggatatag accccaactt ctcaatatat attggtggga 16140ctgcaggtga ccgtggatta tcggacgctg caagattatt tctccgaatt gcaatttcaa 16200ctttcttgag ctttgttgag gagtgggtta tctttaggaa ggcaaacatc ccactatggg 16260ttatctatcc tctcgaaggc caacgctctg atcctcctgg cgaatttttg aaccgagtaa 16320aatctctaat tgttgggact gaagatgata aaaataaagg ctctatactt tcaagatctg 16380gagagaaatg ctcttcaaat ctagtttata attgcaagag tacagcaagc aattttttcc 16440atgcatcatt ggcttactgg agaggtcgac atagacctaa gaagactata ggtgcaacta 16500acgcgacaac agctccacat atcattttgc cactgggaaa ttctgatcga ccgcctggcc 16560tagaccttaa taggaacaat gatactttca ttcctaccag aattaaacag atagtccaag 16620gagactctag aaacgacaga acgaccacca cgagatttcc acccaaaagt aggtccactc 16680caacatcagc aaccgagcct cctacaaaaa tgtatgaggg ttcgacaacc caccaaggga 16740aattaacaga tacacatttg gatgaggatc acaatgccaa agagttccca tccaatccgc 16800atcgtttagt agtaccattc tttaaattaa caaaagatgg ggaatacagc atcgaacctt 16860ctcctgaaga aagccgcagt aatataaaag ggttacttca acatttaaga accatggttg 16920atactaccat atattgtcgc ttcactggaa ttgtttcatc aatgcattat aagttagatg 16980aagtactatg ggaatataat aaatttgaat cagctgtaac cctagcagaa ggggagggtt 17040caggtgcctt actactgatc caaaaatacg gcgttaagaa gttatttttg aatacacttg 17100ctactgaaca tagtattgag agtgaagtga tatcaggtta caccactcca aggatgctac 17160tcccaattat gcctaaaaca catcgtggtg agctagaggt catattaaat aactcagcta 17220gtcaaataac tgatattaca catcgagatt ggttttcaaa tcaaaaaaat aggattccaa 17280atgatgctga tattattacc atggatgctg aaactacaga aaacttagat cgttccagat 17340tatatgaagc agtatatacg attatttgta atcatatcaa tcctaaaact ttgaaagtgg 17400tcatcttaaa agtcttcctc agcgatttgg atgggatgtg ctggattaac aattatcttg 17460ctcctatgtt tggatcagga tatttaatca aacctataac atcaagtgca aagtcaagtg 17520agtggtattt atgcttatct aatctacttt caaccttgag aactactcag catcaaaccc 17580aggcaaactg tctccatgtc gtacaatgtg ctcttcaaca gcaagtacaa agagggtcat 17640attggctaag tcatcttacc aaatacacca caagtagatt gcacaatagt tatattgcat 17700ttggttttcc ttcattagag aaggtcctat atcataggta taaccttgtt gattcgagaa 17760atggaccatt agtttctata acgagacacc ttgccctcct ccaaactgag atccgggagt 17820tggtaactga ttataatcag ctgcgacaaa gtcgaaccca gacttatcat ttcataaaaa 17880catccaaggg acggataact aaactagtga atgattatct aagatttgag ttggttatac 17940gggctcttaa aaataattct acatggcacc atgagttata cttgctacca gaacttatag 18000gtgtttgcca tcgatttaat catacacgta actgtacatg cagtgaaagg ttcctggttc 18060aaactttata tctacaccga atgagtgatg ctgagataaa acttatggac cggctcacca 18120gcctagtcaa tatgtttcct gaaggtttca ggtctagttc agtctaattc taactgcacc 18180aaaggctcta aaaatatttt aaataaccag gtgtatatca aagtcaatac aagtgtaaaa 18240acaatatgca agggaccaca tttaggatca gtttattgac tcttccaata cacagagttg 18300gaagcaccga ttcaaggttt ctaagacgcc ctatcgatta tgttgataat gtaaataata 18360gcttttcctg tctattatga cttaaataat catatctata acgaccatca cagctaagtc 18420gttgccctag ttcatatatt aaattaaaat ttagaagcta ggttgactct aattacataa 18480gtattaagaa aaaattacta agactaatac tctcatgcca agaactagta atgtgtttca 18540catgacagat tatttctaac actaaattgc aatttcaatt ttaaagctaa gtttaacacc 18600tatacagcca aaatatttca tagggccgat gggaataaca taagaggaac atgatcaatg 18660aaccctttat tccaactagg cagttgattg ataatctaca aattccataa gatgttctta 18720cgatattctt ttgtttttaa tctcaatgtc aatgatttaa taagtaataa taaaaaaatc 18780acattaaaga tgcaggaaga tcttgacctc gccaggaaaa ttaagcgcac acaaataaat 18840taaaaaatct gtattttctc ttttttgtgt gtcca 188752251PRTSudan ebolavirus 2Met Ala Lys Ala Thr Gly Arg Tyr Asn Leu Val Thr Pro Lys Arg Glu1 5 10 15 Leu Glu Gln Gly Val Val Phe Ser Asp Leu Cys Asn Phe Leu Val Thr 20 25 30 Pro Thr Val Gln Gly Trp Lys Val Tyr Trp Ala Gly Leu Glu Phe Asp 35 40 45 Val Asn Gln Lys Gly Ile Thr Leu Leu Asn Arg Leu Lys Val Asn Asp 50 55 60 Phe Ala Pro Ala Trp Ala Met Thr Arg Asn Leu Phe Pro His Leu Phe65 70 75 80 Lys Asn Gln Gln Ser Glu Val Gln Thr Pro Ile Trp Ala Leu Arg Val 85 90 95 Ile Leu Ala Ala Gly Ile Leu Asp Gln Leu Met Asp His Ser Leu Ile 100 105 110 Glu Pro Leu Ser Gly Ala Leu Asn Leu Ile Ala Asp Trp Leu Leu Thr 115 120 125 Thr Ser Thr Asn His Phe Asn Met Arg Thr Gln Arg Val Lys Asp Gln 130 135 140 Leu Ser Met Arg Met Leu Ser Leu Ile Arg Ser Asn Ile Ile Asn Phe145 150 155 160 Ile Asn Lys Leu Glu Thr Leu His Val Val Asn Tyr Lys Gly Leu Leu 165 170 175 Ser Ser Val Glu Ile Gly Thr Pro Ser Tyr Ala Ile Ile Ile Thr Arg 180 185 190 Thr Asn Met Gly Tyr Leu Val Glu Val Gln Glu Pro Asp Lys Ser Ala 195 200 205 Met Asp Ile Arg His Pro Gly Pro Val Lys Phe Ser Leu Leu His Glu 210 215 220 Ser Thr Leu Lys Pro Val Ala Thr Pro Lys Pro Ser Ser Ile Thr Ser225 230 235 240 Leu Ile Met Glu Phe Asn Ser Ser Leu Ala Ile 245 250 318891DNAReston ebolavirus 3cggacacaca aaaagaaaaa aggtttttta agactttttg tgtgcgagta actatgagga 60agattaacag ttttcctcag tttaagatat acactgaaat tgagattgag attctcctct 120ttgctattct gtaactttcc ctggttgtga caattgaatc agttttatct attaccaatt 180accatcaaca tggtatgtct agtgatcttg ggactcttct tcatctggtt tttcctagag 240ctctgaatcc attttgcgag aagttcatcc aaacgaccca gtgtctgaaa atacaaaagg 300ttcccctttc cgtcaagttt aaggggttgt tttgattgtg tgtagatttt ataatcctag 360agtgccaagg agttgcgtgt catcattgat tgggaagatc aaggaaacaa tttgttccaa 420taatatcgta catcttgact aagtcgaaca aggggaagtc gatatggatc gtgggaccag 480aagaatctgg gtgtcgcaaa atcaaggtga tactgattta gattatcata aaattttgac 540agctggcctt actgttcaac agggaattgt caggcagaaa ataatttctg tatatcttgt 600tgataacttg gaggctatgt gtcaattggt aatacaagcc tttgaggccg gaattgattt 660ccaagaaaat gccgacagct tccttctgat gctttgccta catcatgctt accaaggtga 720ctataaattg ttcttggaga gcaatgctgt acagtatttg gaaggtcatg gattcaaatt 780tgagctccgg aagaaggacg gtgtcaatcg gctcgaggaa ttgcttcctg ctgcaacgag 840tggaaaaaac atcaggcgta cgttggccgc actgcctgaa gaggagacta cagaagcaaa 900tgcagggcaa tttctctcat ttgcgagttt gtttcttccc aaactggttg tgggagagaa 960ggcttgcttg gaaaaagtcc agcgacaaat tcaggttcat gcagaacagg gtttaattca 1020atatcccact gcatggcaat cagttggaca catgatggta atcttcagat tgatgaggac 1080taatttcttg attaaatatt tactgatcca ccagggtatg catatggtag ctggccacga 1140tgccaatgat gctgtcattg ctaattcagt tgctcaggct cgcttttcag gactcctaat 1200tgtcaaaacc gttcttgatc atattctgca aaaaaccgac caaggagtaa gacttcaccc 1260tttggcccga acagccaaag tgcgtaatga ggttaatgca tttaaggccg ccctaagctc 1320acttgctaag catggggaat atgccccttt tgctcgcctt ctcaatctct cgggagttaa 1380caacctagaa catggtctct acccacagtt atcagcaatt gctcttggag ttgccacagc 1440acatggtagc acccttgcag gagttaatgt tggtgagcag tatcagcagc ttagagaggc 1500tgccactgaa gctgagaagc aactccaaca atatgctgag tccagagaac tcgacagcct 1560aggcctggac gatcaggaaa gaagaatact aatgaacttc catcagaaga aaaacgaaat 1620tagtttccag cagaccaatg caatggtaac ccttaggaaa gagcgactgg ctaaattaac 1680agaagctata acgctggcct caagacctaa cctcgggtct agacaagacg acggcaatga 1740aataccgttc cctgggccta taagcaacaa cccagaccaa gatcatctgg aggatgatcc 1800tagagactcc agagacacca tcattcctaa tggtgcaatt gaccccgagg atggtgattt 1860tgaaaattac aatggctatc atgatgatga agttgggacg gcaggtgact tggtcctgtt 1920cgatcttgac gatcatgagg atgacaataa agcttttgag ccacaggaca gctcgccaca 1980atcccaaagg gaaatagaga gagaaagatt aattcatcca cccccaggca acaacaagga 2040cgacaatcga gcctcagaca acaatcaaca atcagcagat tctgaggaac aaggaggtca 2100atacaactgg caccgaggcc cagaacgtac gaccgccaat cgaagactct caccagtgca 2160cgaagaggac acccttatgg atcaaggtga tgatgatccc tcaagcttac ctccgctgga 2220atctgatgat gacgatgcat caagtagcca acaagatccc gattatacag ctgttgcccc 2280tcctgctcct gtataccgca gtgcagaagc ccacgagcct ccccacaaat cctcgaacga 2340gccagctgaa acatcacaat tgaatgaaga ccctgatatc ggtcaatcaa agtctatgca 2400aaaattagaa gagacatatc accatctgct gagaactcaa ggtccatttg aagccatcaa 2460ttattatcac atgatgaagg atgagccggt aatatttagc actgatgatg ggaaggaata 2520cacctacccg gattcacttg aggaagccta tcctccatgg ctcaccgaga aagaacgact 2580ggacaaagag aatcgctaca tttacataaa taatcaacag ttcttctggc ctgtcatgag 2640tcccagagac aaatttcttg caatcttgca gcaccatcag taaccacagc acaaagcgcg 2700gtccacttcg taaagctaaa tacacttaag acttgaccga ttcatctaca aaaactaatc 2760cattataact tattagtgct acttttctat aagtgattct taatctaagg ccattaagag 2820tttaagtaat atacatatac acttacaccg gtctatccaa gatgtggctc aatgttcttg 2880atttgaacat agtcataagg ggataaataa tactttatat ttctgattgt ggattgaccc 2940attctgctta aaatgcttcg cccattgaaa atgtgatcta atagatagcc ctgactagac 3000aaattaagaa aaacatttga tgaagattaa aaccttcatc gccagtaaat gattatattg 3060tctgtaggca ggtgtttact ccaccttaaa tttggaaata tcctacctta ggaccattgt 3120caagaggtgc ataggcatta ccacccttga gaacatgtac aataataaat tgaaggtatg 3180ttcaggccca gaaacgactg gatggatttc tgagcaactt atgacaggta agattccagt 3240aactgatata ttcattgata ttgataacaa gccagatcaa atggaagtcc gactcaaacc 3300atcatcaagg agctcaacaa gaacttgtac aagtagcagt cagacggagg tcaactatgt 3360acctctcctt aaaaaggttg aggatacatt aactatgcta gtgaatgcca ccagtcgtca 3420gaatgctgca atcgaggccc ttgaaaaccg cctcagcaca cttgagagta gcttaaagcc 3480aatccaagac atgggtaaag tgatttcatc attgaatcgc agttgtgccg aaatggttgc 3540aaaatatgat cttctagtta tgacaactgg acgggctact tcaactgcag ctgcagtaga 3600tgcgtattgg aaagagcaca aacagccacc accagggcca gcgttgtatg aagagaatgc 3660gcttaaagga aaaatcgatg atccaaacag ctatgtacca gatgctgtgc aagaggctta 3720caagaacctt gacagtacat cgaccctgac cgaggaaaat tttgggaaac cttatatatc 3780tgctaaagac ctgaaggaga tcatgtatga tcatctacct ggttttggga ctgcctttca 3840ccaacttgtt caagtgattt gtaaaatagg aaaggataac aaccttttgg acacaatcca 3900tgctgagttc caggcaagtc tagcagatgg tgactctccc caatgtgcac tcatacagat 3960aaccaaaagg gtcccaatct ttcaggatgt gccgcccccg ataatccata ttagatcccg 4020tggtgacatc ccacgagcat gccaaaagag tctccgacca gcaccaccat cacccaaaat 4080tgatcgtggt tgggtttgtt tgtttaagat gcaagatggt aaaacgcttg gacttaagat 4140ctaagaatca agatttattt aacaaggcaa gccacaacct tagatggaac ctcagccaga 4200ctattgaact attgacgctg ttgatgataa tatataatta atggtcttat ttgaatatga 4260caacatcttg cttcttgttc tgccttgtag ctctttgaat tggaagatca ttccaaactt 4320acaaacatgc acaagatgtt atggtttagc aaagaattga taggagtact ggtatataat 4380gtaaatataa caagtgatga agattaagaa aaaccagtcg gtattttcca gacttggcat 4440ttcttatctt catcttctaa agtgagatat tttatcatca aaaaatgagg cgcggagtgt 4500taccaacggc tcctccagca tataatgata ttgcataccc tatgagcata ctcccaaccc 4560gaccaagtgt catagtcaat gagaccaaat cagatgtact ggcagtgcca ggggcagatg 4620ttccatcaaa ctccatgaga ccagtggctg atgataacat tgatcactca agccatactc 4680caagcggagt agcttctgcc tttatattgg aagctacagt gaatgtaatt tcgggaacaa 4740aagtcctgat gaagcaaata cctatttggc ttccactggg tgtagctgat cagaagatat 4800acagctttga ttcaacaaca gccgcaatta tgttggcttc ctacacagtg acacacttcg 4860ggaagatatc taacccgctg gtacgtgtca acaggctagg cccaggaata cccgatcatc 4920cgctacgact cctaaggttg ggcaatcagg cattccttca agagtttgtt cttccaccag 4980tccagcttcc ccagtatttc acatttgatc taacagctct aaagctcatc actcaaccat 5040tgccagctgc aacctggaca gacgaaactc cagcaggagc agtcaatgct cttcgtcctg 5100ggctctcact ccatcccaag cttcgtccaa ttctcctgcc ggggaagaca ggaaagaaag 5160gacatgcttc agacttaaca tcacctgaca agattcaaac aatcatgaat gcaataccgg 5220acctcaaaat tgtcccgatt gatccaacca agaacatagt tggaattgag gttccagaat 5280tactagttca aaggctgacc ggcaaaaaac cacaacccaa aaatggccaa ccaattattc 5340cagttcttct tccgaaatat gttggacttg atcctatatc gccaggggac ttaactatgg 5400ttatcaccca ggattgtgat tcatgccact ctccagccag ccatccgtat cacatggaca 5460agcagaatag ttaccaataa tttaaattcc attcgagcta ttattctgct agtaattccg 5520acgggatcaa tagactaaaa atctgattgt atagaattat aaaagaatca agcagaggca 5580acagactcac agcttacgcc tagataacta atattaagga gttttttaat ctaattttcc 5640agtcttgagt aataatcatt tcttttgtaa ttaattatgc atttgttaac ttatcggtgc 5700gagatttcct tgagaacccg gcggagcttc tactatctgc agtaaccaga agagaagttc 5760aacccagtca aaactaaacc aagcaatatt ctgaatgctc tatagtctat tctaatcaga 5820ggtataacaa tggctaagat ttcaatgact cgttaacaat cgctagtaat tttaatctcc 5880agattaagaa aaagatatac gatgaagatt aaggcgacaa cgagccgaaa cttcatctct 5940tttaaagatc taacattatc tgttccaaag tcatacaagg acacattcaa atcagggatt 6000gtaagctgct atttcttacc tccccaaatt acctatacaa catggggtca ggatatcaac 6060ttctccaatt gcctcgggaa cgttttcgta aaacttcgtt cttagtatgg gtaatcatcc 6120tcttccagcg agcaatctcc atgccgcttg gtatagtgac aaatagcact ctcaaagcaa 6180cagaaattga tcaattggtt tgtcgggaca aactgtcatc aaccagtcag ctcaagtctg 6240tggggctgaa tctggaagga aatggaattg caaccgatgt cccatcagca acaaaacgct 6300ggggatttcg ttcaggtgtg cctcccaagg tggtcagcta tgaagccgga gaatgggcag 6360aaaattgcta caatctggag atcaaaaagt cagacggaag tgaatgcctc cctctccctc 6420ccgacggtgt acgaggattc cctagatgtc gctatgtcca caaagttcaa ggaacaggtc 6480cttgtcctgg tgacttagct ttccataaaa atggggcttt tttcttgtat gatagattgg 6540cctcaactgt catctaccga gggacaactt ttgctgaagg tgtcgtagct tttttaattc 6600tgtcagagcc caagaagcat ttttggaagg ctacaccagc tcatgaaccg gtgaacacaa 6660cagatgattc cacaagctac tacatgaccc tgacactcag ctacgagatg tcaaattttg 6720ggggcaatga aagcaacacc ctttttaagg tagacaacca cacatatgtg caactagatc 6780gtccacacac tccgcagttc cttgttcagc tcaatgaaac acttcgaaga aataatcgcc 6840ttagcaacag tacagggaga ttgacttgga cattggatcc taaaattgaa ccagatgttg 6900gtgagtgggc cttctgggaa actaaaaaaa cttttcccaa caacttcatg gagaaaactt 6960gcatttccaa attccatcaa cccacaccaa caactcctca gatcagagcc cggcgggaac 7020tgtccaagga aaaattagct accacccacc cgccaacaac tccgagctgg ttccaacgga 7080ttcccctcca gtggtttcag tgctcactgc aggacggaca gaggaaatgt cgacccaagg 7140tctaaccaac ggagagacaa tcacaggttt caccgcgaac ccaatgacaa ccaccattgc 7200cccaagtcca accatgacaa gcgaggttga taacaatgta ccaagtgaac agccgaacaa 7260cacagcatcc attgaagact cccccccatc ggcaagcaac gagacaattt accactccga 7320gatggatccg atccaaggct cgaacaactc cgcccagagc ccacagacca agaccacgcc 7380agcacccaca acatccccga tgacccagga cccgcaagag acggccaaca gcagcaaacc 7440aggaaccagc ccaggaagcg cagccggacc aagtcagccc ggactcacta taaatacagt 7500aagtaaggta gctgattcac tgagtcccac caggaaacaa aagcgatcgg ttcgacaaaa 7560caccgctaat aaatgtaacc cagatcttta ctattggaca gctgttgatg agggggcagc 7620agtaggattg gcatggattc catatttcgg acctgcagca gaaggcatct acattgaggg 7680tgtaatgcat aatcagaatg ggcttatttg cgggctacgt cagctagcca atgaaactac 7740ccaggctctt caattatttc tgcgggccac aacagaactg aggacttact cacttcttaa 7800cagaaaagct attgattttc ttcttcaacg atggggaggt acctgtcgaa tcctaggacc 7860atcttgttgc attgagccac atgattggac aaaaaatatt actgatgaaa ttaaccaaat 7920taaacatgac tttattgaca atcccctacc agaccacgga gatgatctta atctatggac 7980aggttggaga caatggatcc cggctggaat tgggattatt ggagttataa ttgctataat 8040agccctactt tgtatatgta agattttgtg ttgatttatt ctgagatctg agagagaaaa 8100atctcagggt tactctaagg agaaatatta tttttaaaat ttacttgaat gctgaccact 8160tatcttaaat gagcaattaa taatatgttt ttctgcttct ttgcttgatt tacaatatga 8220tatttctctt aataatgatt aatatattaa gaaaaactta tgacgaagat taaaggagag 8280gatcgttaac gggaaaacct cccatctcgt tcgtcgaagc cacgttggtg gtgcttgcag 8340ctgagaacaa ctccagagat tgtaggtaga aaggaccaac atttataggt aggggtcaga 8400aagcaacaat aaccataaaa ggagagcctg acattgctat ttaatatcct agaacctgat 8460ttctaggttc tagctttaaa atccggatga tggagcattc aagagaacgg ggtagatcta 8520gcaacatgcg acataatagc cgggaaccat acgaaaatcc atcaaggtct cgctcattat 8580ctcgggaccc taatcaggtt gatcgtagac agcctcgaag tgcatcccaa attcgtgttc 8640cgaatctgtt ccatcggaaa aagactgatg cactcatagt tcctccggct cctaaagata 8700tatgcccaac actcaaaaaa ggattcctct gcgatagcaa attttgcaaa aaagatcacc 8760aattggatag cttaaatgat catgaattac tactgctaat tgcaagaaga acatgtggaa 8820ttatcgagag caattcgcag attacatccc caaaagatat gcggttagcg aatccaacag 8880ctgaagactt ctcacaaggt aatagtccta aattaacact tgcagtcctt cttcaaattg 8940ctgaacattg ggcaaccaga gacctaaggc aaattgagga ctctaaactt agagctcttt 9000taaccctttg tgccgtatta acaaggaaat tttctaaatc ccaactgggt cttctatgtg 9060agacccacct acggcatgag ggcctcggac aggaccaagc tgattctgta ttagaggtct 9120accaaagact ccacagtgat aaaggaggga attttgaggc tgccctgtgg caacaatggg 9180accgacagtc attaataatg ttcatctctg cttttctcaa cattgctctc cagatacctt 9240gtgaaagttc tagtgtcgta gtctcaggtc ttgccacatt gtacccagca caagacaatt 9300ctacaccatc cgaggcaact aatgatacca cctggtcaag tacagttgaa tagaaaacca 9360ctggagctat ttttccacga ttgctctcag tcaataaatt aatatagata taatacgact 9420tcggtgtgca attgtcaaga gttccattta gtaataatga ttcttaaaac aatctactat 9480cgcaattatc gatggatcta ccctatttga cggtacatga

cttgaatgta ataaggtaag 9540ttggtatctg aggtattttg tctagagtat actcaaaatc gtatgtctag caaattatca 9600atagcaaagt taaattctcc taacctcata ttttgatcaa gtaatcatga ttttatgata 9660attcttttca gattatcggt ttaatcttta ttaagaaaaa atcatgattg tagacaattt 9720actggtagtc cttgggtatc caagtttatg aatagagcta gagagaattt gctacttccg 9780aggtataact ttattatttg ctacttcgaa tgcctaaaac cagtaatgca ggatgaagat 9840taattgcgga ggaatcagga attcaacttt agttccttaa ggcctcgtcc gaatcttcat 9900cagttcgtaa gttcttttat agaagtcatt agcttctaag gtgattatat tttagtatta 9960aattttgcta attgcttgct ataaagttga aatgtctaat gcttaaatga acactttttt 10020gaagctgaca tacgaataca tcatatcata tgaaaacatc gcaattagag cgtccttgaa 10080gtctggcatt gacagtcacc aggctgttct cagtagtctg tccttggaag ctcttgggga 10140gacaaaaaga ggtcccagag agtcccaaca ggttggcata aggtcattaa caccagcata 10200gtcggctcga ccaagactgt aagcgagtcg atttcaacta aaaagattat ttcttgttgt 10260ttaaacaaat tccttttgtg tgagacatcc tcaaggcaca agatggctaa agccacaggc 10320cgatacaatc tcgtgccccc aaagaaagat atggaaaagg gagtgatttt tagtgatctt 10380tgtaatttct tgattactca aaccctgcaa ggttggaagg tttattgggc aggaattgag 10440tttgatgtaa gtcaaaaagg catggctctt ctgacaagac tcaaaacaaa tgactttgct 10500cctgcctggg cgatgacaag aaatcttttc ccacatctgt tccagaaccc aaattcggtt 10560attcaatctc ccatctgggc tttgagggta attttggcag ccggattgca ggatcagttg 10620ttagaccatt cattggttga gccattgaca ggggctctcg gtctaatttc tgattggctc 10680ctaactacaa cgtcaacaca tttcaatctt cgtactagaa gcgtaaagga ccagcttagt 10740cttcgtatgt tatctttgat caggtcaaac atcttgcagt tcatcaacaa gcttgacgcc 10800ctgcatgttg tcaattacaa tggtttactc agtagtattg agatcgggac ttctacacac 10860acaatcatta taactcgtac aaatatgggt tttctcgtgg aagttcaaga gcctgacaaa 10920tcagctatga attctaagcg cccaggacca gtcaagttct cattacttca tgagtctgcc 10980ttcaaacctt tcactcgtgt tccacaatct gggatgcaat cattaataat ggagttcaac 11040agtttgttgg caatttaaca aggtaatctt aaaataagta catgaatgag aattagttgt 11100gggtcttatc tagcattgtt gagttaacta tctaatctat tttcgctaat tgcattgagc 11160actgctaata ggtttgtatc acgttaaaga tttagagtgt atgaattgtg cagatttaaa 11220cttgggtttt gccttatgct tcataggtgg tctttttgaa atggagatta tcagcatttc 11280ttaaatggga ggagttagca atcagaaatt ggagataaat ggacatcggg atagaacaat 11340gcctaactat tgggcggctt ccatttttac atgtgtatat aaccaatctt ttcctatctt 11400tgcttatatt ggtgtaactt tattttaata acatgtcaat gctatactgt taagagaagg 11460tctgaggaag attaagaaaa aggcctcgtg ttcacttggt tgccgtcaag tatcctgtgg 11520tttttttcta cctaacttcc tcatgccata tggctaccca gcatacccag tacccggatg 11580cacgtttatc ctcacctata gtcctggatc aatgtgattt ggtaactcga gcatgtgggt 11640tatattcatc ttattctcta aatcctcaac taaggcaatg taaattacca aaacatatat 11700atcgacttaa gttcgacaca atagtatcca aattcctaag tgatacacct gtagcaacac 11760tgccgataga ctatttagta ccaattctcc tgcgttccct aacggggcac ggtgataggc 11820cattgacccc gacttgcaat caattccttg atgaaattat taattacact cttcatgatg 11880cagcctttct tgattactat ctcaaggcaa caggtgcaca ggaccatttg acaaacattg 11940caactagaga gaagcttaaa aacgaaattc taaacaatga ttatgtccat caattgttct 12000tctggcatga cctttctatt ttggctcgac gtgggcgtct gaatcgcggg aacaaccgtt 12060caacctggtt tgttcatgat gaattcattg atattttagg atatggcgat tatatttttt 12120ggaaaatacc tttatcatta ttaccagtta ctatagacgg ggtcccacac gcagcaactg 12180actggtatca accgactctt tttaaagaat ccatcctagg gcatagccaa atcctatctg 12240tgtcaacagc tgaaatacta attatgtgta aagatattat cacctgtagg tttaatacat 12300cactgattgc atccattgca aaattagagg atgtagatgt gtctgattat cctgacccga 12360gtgatattct taagatatac aatgctggag actatgtaat atctattctt ggctcagagg 12420gttataagat aataaagtac cttgaaccac tttgtttggc caaaatccaa ctttgctcta 12480aattcacaga aagaaaaggt cgtttcctca cacagatgca tttatcagta ataaatgatc 12540ttcgggagtt gatttctaac cgcaggttaa aggactatca gcaagagaag attagagatt 12600ttcacaaaat attattacaa ttgcaattat ctcctcaaca gttttgtgaa ttattctctg 12660ttcaaaaaca ttgggggcat ccaattttac atagtgagaa agctatacaa aaagtaaaac 12720ggcatgcaac catccttaag gctctcagac ctaatgtcat ctttgagaca tattgtgtat 12780tcaagtacaa tattgccaag cactatttcg acagccaagg aacttggtac agtgtaatct 12840cagacaggaa tttaactcca ggactcaact ccttcataaa acgtaatcac tttccttcac 12900tacccatgat taaggatctt ctatgggaat tctatcatct taatcaccct ccgttattct 12960ctacaaaggt gattagtgac ttaagtattt tcatcaaaga tagggccaca gctgttgaac 13020agacatgttg ggatgcagtc tttgaaccca atgtgctagg ttacaatcct ccaaacaaat 13080tctccactaa aagggtgccg gaacaatttc tagaacaaga ggatttttca atcgaaagtg 13140tcctgaatta tgcacaggaa ttacattatt tattaccaca gaataggaat ttttcctttt 13200ctctcaaaga aaaggaatta aatattggac gaacatttgg gaagctacca tatctcacac 13260ggaatgtcca aactttatgt gaggctctgt tagcagatgg actggccaag gccttcccca 13320gtaacatgat ggtagtaact gaacgtgaac aaaaagagag ccttcttcat caggcatcat 13380ggcaccacac cagtgatgat tttggagaga atgctaccgt tcgagggagt agttttgtaa 13440ctgatttaga gaagtacaat cttgcatttc gctatgagtt cactgcacca tttattgagt 13500actgcaacca ttgctatggt gtgcgtaatg tctttaattg gatgcattat ttaatcccgc 13560agtgttacat gcatgtaagt gattattata atccgcctca caatgttaat cttagcaatc 13620gagaatatcc tcctgaaggc ccgagttcgt accgagggca cttaggaggc atagagggat 13680tacaacaaaa actgtggacg agtatatcct gtgcacaaat ctccttagtg gaaattaaaa 13740ctggttttaa gttacgatca gcggtcatgg gagacaatca gtgtataacc gtattgtctg 13800tttttccact taaaacagac cctgaagagc aggagcaaag cgccgaagac aatgctgcaa 13860gagtagcagc aagtcttgca aaagtaacca gtgcatgtgg gatctttctt aaaccagatg 13920agacatttgt acactcaggt ttcatttatt tcggaaaaaa acaatatctc aatggtgtac 13980aattaccgca atcactcaaa acagcagcaa gaatggcacc actctctgat gctatattcg 14040atgatctaca aggaacactt gccagtattg gaactgcctt cgaacgtgct atatcggaaa 14100cgcgacatat cctcccatgt cgtattgtag cagctttcca tacgtatttc gccgttcgga 14160ttttacaata tcaccatctt ggatttaata aaggcatcga tttaggacag ttgtcactta 14220gtaaaccatt agactatggg actattactc taacattggc ggttccacaa gtccttgggg 14280gattgtcttt tctaaatcca gaaaagtgtt tttatcgaaa cttcggagat cctgtgactt 14340ctggactttt ccagctacgg gtgtacctag aaatggttaa catgaaagac ctattttgtc 14400cattaatatc gaaaaatcca ggaaattgta gtgccattga ttttgtctta aatccatccg 14460gattaaatgt tccaggatca caagacttga catccttttt gcgacaaatc gttaggcgta 14520gtattaccct aactgctaga aataagttaa ttaacactct cttccatgcc tctgctgatt 14580tggaagatga gatggtttgt aagtggctcc tttcatcaaa ccctgtcatg agtcgctttg 14640cagcggatat tttttccagg acaccgagtg gtaaacgtct ccaaatatta ggttatcttg 14700aagggaccag gactctattg gcctccaaaa tcataaacaa caacagtgag acacctgtac 14760ttgataagct gaggaagatc accctacaaa gatggaatct gtggttcagt tatttggacc 14820attgtgacca attactagca gatgctctac agaaaattag ttgcacggtg gatttggccc 14880agattttgcg tgagtataca tggtcacaca tcttagaggg tagatcattg atcggagcga 14940cattaccatg tatggtggag caattcaaag ttaagtggct aggacaatat gaaccttgtc 15000cagaatgtct caacaaaaaa ggctcaaatg cttatgtctc agttgcagtc aaagatcaag 15060tggtcagtgc ttggcctaat acttctcgaa taagttggac aatagggagt ggtgtcccct 15120atatagggtc aagaaccgag gataaaatcg gacagcctgc tatcaagccg cgatgccctt 15180catctgccct caaggaggct atagaattag catcaaggct cacttgggtt acacaaggag 15240gttctaatag tgaacaatta atccggcctt tcttggaagc gagagtcaac cttagtgtca 15300gtgaagtcct gcaaatgaca ccatcacatt attcaggaaa tattgtccat cgatataacg 15360accaatacag cccgcactca tttatggcga atcgcatgag caatactgcg acccgtctca 15420tagtgtcaac taatacactt ggagaatttt caggtggagg gcaggccgcc agggatagca 15480atataatttt ccagaatgtt ataaatttag cagttgccct ttatgatatt agattccgga 15540atacaaacac ctctgatata aggcataata gggctcatct tcacctgaca gagtgctgta 15600ctaaagaggt cccggcccag tatttgacat atacaagtgc acttaatctg gatttaagcc 15660gttatcgtga taatgaacta atatatgact caaatccact gaagggagga ttgaactgca 15720atttaacaat agatagtcct ttagtgaagg gtcctaggct taacatgatt gaagatgatc 15780ttctccgctt tccacacctt tctggatggg agttagcgaa aacggtggta caatccatca 15840tctcagacaa tagcaactca tcaacagatc caatcagtag cggagaaaca cgctctttca 15900caactcattt tctcacttac cctcagattg gccttcttta cagtttcgga gcagtattat 15960gcttttatct aggcaatact atcctatgga ctaaaaaact tgattacgaa cagtttctat 16020attatttgca taaccagctg cacaacttac ctcatcgagc actccgtgtt tttaaaccaa 16080catttaagca tgccagtgtg atgtcccgat taatggaaat tgattctaac ttctcaattt 16140atattggcgg gacatctgga gatcgagggc tgtctgatgc tgctcgactg tttcttcgga 16200cagcaatcgc gagtttttta caatttctta aaagctggat catcgatcgc caaaagacaa 16260ttcctttatg gatagtatat ccgcttgaag gtcaacagcc ggaatccatc aatgaatttc 16320tacataaaat tttgggtctg ctcaaacaag gccccaaaag tattccaaag gaggtcagca 16380tccaaaatga tggacatttg gatttggcag aaaataatta tgtttacaat agtaagagca 16440ctgctagtaa tttcttccat gcatccttag cttactggag aagtaggaaa tctcggaaaa 16500ctcaagacca taatgatttc tcaagagggg atggaacact tacagaaccc gtgcgtaagt 16560tttcaagcaa tcatcagtca gatgaaaagt actacaatgt gacatgtgga aagtcaccga 16620agccgcaaga acgcaaagac ttctcgcaat acagactcag caataacggg caaacaatga 16680gtaatcatcg taagaaaggg aagttccaca agtggaatcc ctgcaaaatg ttaatggaga 16740gtcaaagggg aactgttcta acagagggtg actactttca aaacaatact ccaccaacag 16800atgatgtatc aagtcctcac cgactcattc taccattttt taaattggga aatcacaacc 16860atgcacatga tcaagatgcc caagaattga tgaatcaaaa tattaaacag tacctacatc 16920agctaaggtc tatgttggac accactatat attgtagatt cacagggatt gtctcatcca 16980tgcattacaa attggacgaa gttcttctag aatacaatag tttcgattca gctatcacat 17040tagctgaagg tgaggggtca ggggctctat tacttttgca aaaatatagt acaaggttat 17100tatttttgaa cacattggca acagaacaca gtatagaatc agaagttgta tcaggttttt 17160ctactccgag aatgttgtta ccaataatgc aaaaggttca tgaaggacaa gtcactgtta 17220tcttaaataa ttcagcaagt cagataactg acataactag ctcaatgtgg ttaagtaatc 17280aaaaatataa tctaccttgt caagttgaaa tcattatgat ggatgctgaa acaacagaga 17340acttaaacag gtcccaactc taccgagcag tatataactt aatacttgat cacattgatc 17400cgcagtatct caaggtggtg gtactcaaag tatttctgag tgatatagaa ggaatattat 17460ggattaatga ttacttggct ccattattcg gggctggtta cttgattaaa ccgattacat 17520caagtgcccg gtcaagtgaa tggtaccttt gcttatcaaa tttgatatct actaacagga 17580gatcggccca tcagactcac aaggcatgtc ttggtgttat cagagatgct ttgcaagcac 17640aagtccagcg aggcgtgtac tggttgagtc acatcgcaca gtatgctaca aagaatctcc 17700attgtgaata cataggcctt ggtttcccat ctctagaaaa ggtcctatat cacaggtata 17760atctagttga tactggactc ggtccattgt cgtcagttat tagacattta actaacctcc 17820aggcagagat acgagactta gtattagatt ataacctgat gagggagagt cgcactcaaa 17880cgtaccattt tattaagact gcaaaaggca gaatcacaaa gttagtcaat gactttctga 17940agttttcttt aattgtccag gcactcaaaa ataattcttc ttggtatact gagcttaaaa 18000aattacctga ggttattaat gtgtgtaatc gattttatca tactcacaat tgcgaatgtc 18060aggaaaaatt ctttgtccag acgctttatt tacaacgcct acgcgatgca gaaatcaagc 18120taattgaacg ccttaccggg ttaatgcgat tttatccaga agggttaata tattccaatc 18180acacataggt actaaatcat catagtatga ggaataagat aatgataatt cctgacgaca 18240gttttagttc cgattctaag tatatcggaa gagagtatgc caatcttaat tgttagaggt 18300aacaagctat tagttattac ttattgataa gaatacactt tatcatagcg taacacatca 18360taactttata acgattttgc atttctaatc ctagtattta ttagaatgta ctaccagaga 18420aatgacccca gttcctatct ttaaataatg attgtgtgta ttaaattatt agtttattag 18480gtttatgagt tggttacaca gtgagtatta gtaattgagg attatgtaga taggtaatct 18540aacactgaat cacccatctg atgtcaccat atccaaatgt tgtgctagtc gcatttaaac 18600atgctatctt cagttaagta acatagactg aaaatgctaa gaagagattg gagtaaaagt 18660ataaaataaa tttaattaaa cttcaaagtg attaaatgat aatgatcttg ggaactcgat 18720atgacctcaa gtcaaaaata atgtcaatat aattgtttag taatatgagt gataatgtaa 18780attttgataa ctaactagct ttagtagtta agatcaaatg caaacattat aagaatgtta 18840agcgcacaca aaaacattat aaaaaaccaa ttttttcctt tttgtgtgtc c 188914287PRTReston ebolavirus 4Met Glu His Ser Arg Glu Arg Gly Arg Ser Ser Asn Met Arg His Asn1 5 10 15 Ser Arg Glu Pro Tyr Glu Asn Pro Ser Arg Ser Arg Ser Leu Ser Arg 20 25 30 Asp Pro Asn Gln Val Asp Arg Arg Gln Pro Arg Ser Ala Ser Gln Ile 35 40 45 Arg Val Pro Asn Leu Phe His Arg Lys Lys Thr Asp Ala Leu Ile Val 50 55 60 Pro Pro Ala Pro Lys Asp Ile Cys Pro Thr Leu Lys Lys Gly Phe Leu65 70 75 80 Cys Asp Ser Lys Phe Cys Lys Lys Asp His Gln Leu Asp Ser Leu Asn 85 90 95 Asp His Glu Leu Leu Leu Leu Ile Ala Arg Arg Thr Cys Gly Ile Ile 100 105 110 Glu Ser Asn Ser Gln Ile Thr Ser Pro Lys Asp Met Arg Leu Ala Asn 115 120 125 Pro Thr Ala Glu Asp Phe Ser Gln Gly Asn Ser Pro Lys Leu Thr Leu 130 135 140 Ala Val Leu Leu Gln Ile Ala Glu His Trp Ala Thr Arg Asp Leu Arg145 150 155 160 Gln Ile Glu Asp Ser Lys Leu Arg Ala Leu Leu Thr Leu Cys Ala Val 165 170 175 Leu Thr Arg Lys Phe Ser Lys Ser Gln Leu Gly Leu Leu Cys Glu Thr 180 185 190 His Leu Arg His Glu Gly Leu Gly Gln Asp Gln Ala Asp Ser Val Leu 195 200 205 Glu Val Tyr Gln Arg Leu His Ser Asp Lys Gly Gly Asn Phe Glu Ala 210 215 220 Ala Leu Trp Gln Gln Trp Asp Arg Gln Ser Leu Ile Met Phe Ile Ser225 230 235 240 Ala Phe Leu Asn Ile Ala Leu Gln Ile Pro Cys Glu Ser Ser Ser Val 245 250 255 Val Val Ser Gly Leu Ala Thr Leu Tyr Pro Ala Gln Asp Asn Ser Thr 260 265 270 Pro Ser Glu Ala Thr Asn Asp Thr Thr Trp Ser Ser Thr Val Glu 275 280 285 518895DNAReston ebolavirus 5cggacacaca aaaagaaaaa aggtttttta agactttttg tgtgcgagta actatgagga 60agattaacag ttttcctcag tttaagatat acactgaaat tgagattgag attctcctct 120ttgctattct gtaactttcc ctggttgtga caattgaatc agttttatct attaccaatt 180accatcaaca tggtatgtct agtgatcttg ggactcttct tcatctggtt tttcctagag 240ctctgaatcc atttcgcgag aagttcatcc aaacgaccca gtgtctgaaa atacaaaagg 300ttcccctttc cgtcaagttt aaggggttgt tttgattgtg tgtagatttt ataatcctag 360agtgccaagg agttgcgtgt catcattgat tgggaagatc aaggaaacaa tttgttccaa 420taatatcgta catcttgact aagtcgaaca cggggaagtc gatatggatc gtgggaccag 480aagaatctgg gtgtcgcaaa atcaaggtga tactgattta gattatcata aaattttgac 540agctgtcctt actgttcaac agggaattgt caggcagaaa ataatttctg tatatcttgt 600tgataacttg gaggctatgt gtcaattggt aatacaagcc tttgaggccg gaattgattt 660ccgagaaaat gccgacagct tccttctgat gctttgccta catcatgctt accaaggtga 720ctataaattg ttcttggaga gcaatgctgt acagtatttg gaaggtcatg gattcaaatt 780tgagctccgg aagaaggacg gtgtcaatcg gctcgaggaa ttgcttcctg ctgcaacgag 840tggaaaaaac atcaggcgta cgttggccgc actgcctgaa gaggagacta cagaagcaaa 900tgcagggcaa tttctctcat ttgcgagtct gtttcttccc aaactggttg tgggagagaa 960ggcttgcttg gaaaaagtcc agcgacaaat tcaggttcat gcagaacagg gtttaattca 1020atatcccact gcatggcaat cagttggaca catgatggta atcttcagat tgatgaggac 1080taatttcttg attaaatatt tactgatcca ccagggtatg catatggtag ctggccacga 1140tgccaatgat gctgtcattg ctaattcagt tgctcaggct cgcttttcag gactcctaat 1200tgtcaaaacc gttcttgatc atattctgca aaaaaccgac caaggagtaa gacttcaccc 1260tttggcccga acagccaaag tgcgtaatga ggttaatgca tttaaggccg ccctaagctc 1320acttgctaag catggggaat atgccccttt tgctcgcctt ctcaatctct cgggagttaa 1380caacctagaa catggtctct acccacagtt atcagcaatt gctcttggag ttgccacagc 1440acatggtagc acccttgcag gagttaatgt tggtgagcag tatcagcagc ttagagaggc 1500tgccactgaa gctgagaagc aactccaaca atatgctgag tccagagaac tcgacagcct 1560aggcctggac gatcaggaag gaagaatact aatgaacttc catcagaaga aaaacgaaat 1620tagtttccag cagaccaatg caatggtaac ccttaggaaa gagcgactgg ctaaattaac 1680agaagctata acgctggcct caagacctaa cctcgggtct agacaagacg acggcaatga 1740aataccgttc cctgggccta taagcaacaa cccagaccaa gatcatctgg aggatgatcc 1800tagagactcc agagacacca tcattcctaa tggtgcaatt gaccccgagg atggtgattt 1860tgaaaattac aatggctatc atgatgatga agttgggacg gcaggtgact gggtcctgtt 1920cgatcttgac gatcatgagg atgacaataa agcttttgag ccacaggaca gctcgccaca 1980atcccaaagg gaaatagaga gagaaagatt aactcatcca cccccaggca acaacaagga 2040cgacaatcga gcctcagaca acaatcaaca atcagcagat tctgaggaac aaggaggtca 2100atacaactgg caccgaggcc cagaacgtac gaccgccaat cgaagactct caccagtgca 2160cgaagaggac acccttatgg atcaaggtga tgatgatccc tcaagcttac ctccgctgga 2220atctgatgat gacgatgcat caagtagcca acaagatccc gattatacag ctgttgcccc 2280tcctgctcct gtataccgca gtgcagaagc ccacgagcct ccccacaaat cctcgaacga 2340gccagctgaa acatcacaat tgaatgaaga ccctgatatc ggtcaatcaa agtctatgca 2400aaaattagaa gagacatatc accatctgct gagaactcaa ggtccatttg aagccatcaa 2460ttattatcac atgatgaagg atgagccggt aatatttagc actgatgatg ggaaggaata 2520cacctacccg gattcacttg aggaagccta tcctccatgg ctcaccgaga aagaacgact 2580ggacaaagag aatcgctaca tttacataaa taatcaacag ttctcctggc ctgtcatgag 2640tcccagagac aaatttcttg caatcttgca gcaccatcag taaccacagc acaaagcgcg 2700gtccacttcg taaagctaaa tacacttaag acttgaccga ttcatctaca aaaactaatc 2760cattataact tattagtgct acttttctat aagtgattct taatctaagg ccattaagag 2820tttaagtaat atacatatac acttacaccg gtctatccaa gatgtggctc aatgttcttg 2880atttgaacat agtcataagg ggataaataa tactttatat ttctgattgt ggattgaccc 2940attctgctta aaatgcttcg cccattgaaa atgtgatcta atagatagcc ctgactagac 3000aaattaagaa aaacatttga tgaagattaa aaccttcatc gccagtaaat gattatattg 3060tctgtaggca ggtgtttact ccaccttaaa tttggaaata tcctacctta ggaccattgt 3120caagaggtgc ataggcatta ccacccttga gaacatgtac aataataaat tgaaggtatg 3180ttcaggccca gaaacgactg gatggatttc tgagcaactt atgacaggta agattccagt 3240aactgatata ttcattgata ttgataacaa gccagatcaa atggaagtcc gactcaaacc 3300atcatcaagg agctcaacaa gaacttgtac aagtagcagt cagacggagg tcaactatgt 3360acctctcctt aaaaaggttg aggatacatt aactatgcta gtgaatgcca ccagtcgtca 3420gaatgctgca atcgaggccc ttgaaaaccg cctcagcaca cttgagagta gcttaaagcc 3480aatccaagac atgggtaaag tgatttcatc attgaatcgc agttgtgccg aaatggttgc 3540aaaatatgat cttctagtta tgacaactgg acgggctact tcaactgcag ctgcagtaga 3600tgcgtattgg aaagagcaca aacagccacc accagggcca gcgttgtatg aagagaatgc 3660gcttaaagga aaaatcgatg atcctaacag ctatgtacca gatgctgtgc aagaggctta 3720caagaacctt gacagtacat cgaccctgac cgaggaaaat tttgggaaac

cttatatatc 3780tgctaaagac ctgaaggaga tcatgtatga tcatctacct ggttttggga ctgcctttca 3840ccaacttgtt caagtgattt gtaaaatagg aaaggataac aaccttttgg acacaatcca 3900tgctgagttc caggcaagtc tagcagatgg tgactctccc caatgtgcac tcatacagat 3960aaccaaaagg gtcccaatct ttcaggatgt gccgcccccg ataatccata ttagatcccg 4020tggtgacatc ccacgagcat gccaaaagag tctccgacca gcaccaccat cacccaaaat 4080tgatcgtggt tgggtttgtt tgtttaagat gcaagatggt aaaacgcttg gacttaagat 4140ctaagaatca agatttattt aacaaggcaa gccacaacct tagatggaac ctcagccaga 4200ctattgaact attgacgctg ttgatgataa tatataatta atggtcttat ttgaatatga 4260caacatcttg cttcttgttc tgccttgtag ctctttgaat tggaagatca ttccaaactt 4320acaaacatgc acaagatgtt atggtttagc aaagaattga taggagtact ggtatataat 4380gtaaatataa caagtgatga agattaagaa aaaccagtcg gtattttcca gacttggcat 4440ttcttatctt catcttctaa agtgagatat tttatcatca aaaaatgagg cgcggagtgt 4500taccaacggc tcctccagca tataatgata ttgcataccc tatgagcata ctcccaaccc 4560gaccaagtgt catagtcaat gagaccaaat cagatgtact ggcagtgcca ggggcagatg 4620ttccatcaaa ctccatgaga ccagtggctg atgataacat tgatcactca agccatactc 4680caagcggagt agcttctgcc tttatattgg aagctacagt gaatgtaatt tcgggaacaa 4740aagtcctgat gaagcaaata cctatttggc ttccactggg tgtagctgat cagaagatat 4800acagctttga ttcaacaaca gccgcaatta tgttggcttc ctacacagtg acacacttcg 4860ggaagatatc taacccgctg gtacgtgtca acaggctagg cccaggaata cccgatcatc 4920cgttccggct cctgaggttg ggcaaaaaag cgttccttcc cgggtttgtt cttccaccag 4980tccagcttcc ccagtatttc acatttgatc taacagctct aaagctcatc actcaaccat 5040tgccagctgc aacctggaca gacgaaactc cagcaggagc agtcaatgct cttcgtcctg 5100ggctctcact ccatcccaag cttcgtccaa ttctcctgcc ggggaagaca ggaaagaaag 5160gacatgcttc agacttaaca tcacctgaca agattcaaac aatcatgaat gcaataccgg 5220acctcaaaat tgtcccgatt gatccaacca agaacatagt tggaattgag gttccagaat 5280tactagttca aaggctgacc ggcaaaaaac cacaacccaa aaatggccaa ccaattattc 5340cagttcttct tccgaaatat gttggacttg atcctatatc gccaggggac ttaactatgg 5400ttatcaccca ggattgtgat tcatgccact ctccagccag ccatacgtat cacatggaca 5460agcagaatag ttaccaataa tttaaattcc attcgagcta ttattctgct agtaattccg 5520acgggatcaa tagactaaaa atctgattgt atagaattat aaaagaatca agcagaggca 5580acagactcac agcttacgcc tagataacta atattaagga gttttttaat ctaattttcc 5640agtcttgagt aataatcatt tcttttgtaa ttaattatgc atttgttaac ttatcggtgc 5700gagatttcct tgagaacccg gcggagcttc tactatctgc agtaaccaga agagaagttc 5760aacccagtca aaactaaacc aagcaatatt ctgaatgctc tatagtctat tctaatcaga 5820ggtataacaa tggctaagat ttcaatgact cgttaacaat cgctagtaat tttaatctcc 5880agattaagaa aaagatatac gatgaagatt aaggcgacaa cgagccgaaa cttcatctct 5940tttaaagatc taacattatc tgttccaaag tcatacaagg acacattcaa atcagggatt 6000gtaagctgct atttcttacc tccccaaatt acctatacaa catggggtca ggatatcaac 6060ttctccaatt gcctcgggaa cgttttcgta aaacttcgtt cttagtatgg gtaatcatcc 6120tcttccagcg agcaatctcc atgccgcttg gtatagtgac aaatagcact ctcaaagcaa 6180cagaaattga tcaattggtt tgtcgggaca aactgtcatc aaccagtcag ctcaagtctg 6240tggggctgaa tctggaagga aatggaattg caaccgatgt cccatcagca acaaaacgct 6300ggggatttcg ttcaggtgtg cctcccaagg tggtcagcta tgaagccgga gaatgggcag 6360aaaattgcta caatctggag atcaaaaagt cagacggaag tgaatgcctc cctctccctc 6420ccgacggtgt acgaggattc cctagatgtc gctatgtcca caaagttcaa ggaacaggtc 6480cttgtcctgg tgacttagct ttccataaaa atggggcttt tttcttgtat gatagattgg 6540cctcaactgt catctaccga gggacaactt ttgctgaagg tgtcgtagct tttttaattc 6600tgtcagagcc caagaagcat ttttggaagg ctacaccagc tcatgaaccg gtgaacacaa 6660cagatgattc cacaagctac tacatgaccc tgacactcag ctacgagatg tcaaattttg 6720ggggcaatga aagtaacacc ctttttaagg tagacaacca cacatatgtg caactagatc 6780gtccacacac tccgcagttc cttgttcagc tcaatgaaac acttcgaaga aataatcgcc 6840ttagcaacag tacagggaga ttgacttgga cattggatcc taaaattgaa ccagatgttg 6900gtgagtgggc cttctgggaa actaaaaaaa cttttcccaa caacttcatg gagaaaactt 6960gcatttccaa attctatcaa cccacaccaa caactcctca gatcagagcc cggcgggaac 7020tgtccaagga aaaattagct accacccacc cgccaacaac tccgagctgg ttccaacgga 7080ttcccctcca gtggtttcag tgctcactgc aggacggaca gaggaaatgt cgacccaagg 7140tctaaccaac ggagagacaa tcacaggttt caccgcgaac ccaatgacaa ccaccattgc 7200cccaagtcca accatgacaa gcgaggttga taacaatgta ccaagtgaac aaccgaacaa 7260cacagcatcc attgaagact cccccccatc ggcaagcaac gagacaattt accactccga 7320gatggatccg atccaaggct cgaacaactc cgcccagagc ccacagacca agaccacgcc 7380agcacccaca acatccccga tgacccagga cccgcaagag acggccaaca gcagcaaacc 7440aggaaccagc ccaggaagcg cagccggacc aagtcagccc ggactcacta taaatacagt 7500aagtaaggta gctgattcac tgagtcccac caggaaacaa aagcgatcgg ttcgacaaaa 7560caccgctaat aaatgtaacc cagatcttta ctattggaca gctgttgatg agggggcagc 7620agtaggattg gcatggattc catatttcgg acctgcagca gaaggcatct acattgaggg 7680tgtaatgcat aatcagaatg ggcttatttg cgggctacgt cagctagcca atgaaactac 7740ccaggctctt caattatttc tgcgggccac aacagaactg aggacttact cacttcttaa 7800cagaaaagct attgattttc ttcttcaacg atggggaggt acctgtcgaa tcctaggacc 7860atcttgttgc attgagccac atgattggac aaaaaatatt actgatgaaa ttaaccaaat 7920taaacatgac tttattgaca atcccctacc agaccacgga gatgatctta atctatggac 7980aggttggaga caatggatcc cggctggaat tgggattatt ggagttataa ttgctataat 8040agccctactt tgtatatgta agattttgtg ttgatttatt ctgagatctg agagagaaaa 8100atctcagggt tactctaagg agaaatatta tttttaaaat ttacttgaat gctgaccact 8160tatcttaaat gagcaattaa taatatgttt ttctgcttct ttgcttgatt tacaatatga 8220tatttctctt aataatgatt aatatattaa gaaaaactta tgacgaagat taaaggagag 8280gatcgttaac gggaaaacct cccatctcgt tcgtcgaagc cacgttggtg gtgcttgcag 8340ctgagaacaa ctccagagat tgtaggtaga aaggaccaac atttataggt aggggtcaga 8400aagcaacaat aaccataaaa ggagagcctg acattgctat ttaatatcct agaacctgat 8460ttctaggttc tagctttaaa atccggatga tggagcattc aagagaacgg ggtagatcta 8520gcaacatgcg acataatagc cgggaaccat acgaaaatcc atcaaggtct cgctcattat 8580ctcgggaccc taatcaggtt gatcgtagac agcctcgaag tgcatcccaa attcgtgttc 8640cgaatctgtt ccatcggaaa aagactgatg cactcatagt tcctccgact cctaaagata 8700tatgcccaac actcaaaaaa ggattcctct gcgatagcaa attttgcaaa aaagatcacc 8760aattggatag cttaaatgat catgaattac tactgctaat tgcaagaaga acatgtggaa 8820ttatcgagag caattcgcag attacatccc caaaagatat gcggttagcg aatccaacag 8880ctgaagactt ctcacatggt aatagtccta aattaacact tgcagtcctt cttcaaattg 8940ctgaacattg ggcaaccaga gacctaaggc aaattgagga ctctaaactt agagctcttt 9000taaccctttg tgccgtatta acaaggaaat tttctaaatc ccaactgggt cttctatgtg 9060agacccacct acggcatgag ggcctcggac aggaccaagc tgattctgta ttagaggtct 9120accaaagact ccacagtgat aaaggaggga attttgaggc tgccctgtgg caacaatggg 9180accgacagtc attaataatg ttcatctctg cttttctcaa cattgctctc cagatacctt 9240gtgaaagttc tagtgtcgta gtctcaggtc ttgccacatt gtacccagca caagacaatt 9300ctacaccatc cgaggcaact aatgatacca cctggtcaag tacagttgaa tagaaaacca 9360ctggagctat ttttccacga ttgctctcag tcaataaatt aatatagata taatacgact 9420tcggtgtgca attgtcaaga gttccattta gtaataatga ttcttaaaac aatctactat 9480cgcaattatc gatggatcta ccctatttga cggtacatga cttgaatgta ataaggtaag 9540ttggtatctg aggtattttg tctagagtat actcaaaatc gtatgtctag caaattatca 9600atagcaaagt taaattctcc taacctcata ttttgatcaa gtaatcatga ttttatgata 9660attcttttca gattatcggt ttaatcttta ttaagaaaaa atcatgattg tagacaattt 9720actggtagtc cttgggtatc caagtttatg aatagagcta gagagaattt gctacttccg 9780aggtataact ttattatttg ctacttcgaa tgcctaaaac cagtaatgca ggatgaagat 9840taattgcgga ggaatcagga attcaacttt agttccttaa ggcctcgtcc gaatcttcat 9900cagttcgtaa gttcttttat agaagtcatt agcttctaag gtgattatat tttagtatta 9960aattttgcta attgcttgct ataaagttga aatgtctaat gcttaaatga acactttttt 10020gaagctgaca tacgaataca tcatatcata tgaaaacatc gcaattagag cgtccttgaa 10080gtctggcatt gacagtcacc aggctgttct cagtagtctg tccttggaag ctcttgggga 10140gacaaaaaga ggtcccagag agtcccaaca ggttggcata aggtcattaa caccagcata 10200gtcggctcga ccaagactgt aagcgggtcg atttcaacta aaaagattat ttcttgttgt 10260ttaaacaaat tccttttgtg tgagacatcc tcaaggcaca agatggctaa agccacaggc 10320cgatacaatc tcgtgccccc aaagaaagat atggaaaagg gagtgatttt tagtgatctt 10380tgtaatttct tgattactca aaccctgcaa ggttggaagg tttattgggc aggaattgag 10440tttgatgtaa gtcaaaaagg catggctctt ctgacaagac tcaaaacaaa tgactttgct 10500cctgcctggg cgatgacaag aaatcttttc ccacatctgt tccagaaccc aaattcggtt 10560attcaatctc ccatctgggc tttgagggta attttggcag ccggattgca ggatcagttg 10620ttagaccatt cattggttga gccattgaca ggggctctcg gtctaatttc tgattggctc 10680ctaactacaa cgtcaacaca tttcaatctt cgtactagaa gcgtaaagga ccagcttagt 10740cttcgtatgt tatctttgat caggtcaaac atcttgcagt tcatcaacaa gcttgacgcc 10800ctgcatgttg tcaattacaa tggtttactc agtagtattg agatcgggac ttctacacac 10860acaatcatta taactcgtac aaatatgggt tttctcgtgg aagttcaaga gcctgacaaa 10920tcagctatga attctaagcg cccaggacca gtcaagttct cattacttca tgagtctgcc 10980ttcaaacctt tcactcgtgt tccacaatct gggatgcaat cattaataat ggagttcaac 11040agtttgttgg caatttaaca aggtaatctt aaaataagta catgaatgag aattagttgt 11100gggtcttatc tagcattgtt gagttaacta tctaatctat tttcgctaat tgcattgagc 11160actgctaata ggtttgtatc acgttaaaga tttggagtgt atgaattgtg cagatttaaa 11220cttgggtttt gccttatgct tcataggtgg tctttttgaa atggagatta tcagcatttc 11280ttaaatggga ggagttagca atcagaaatt ggagataaat ggacatcggg atagaacaat 11340gcctaactat tgggcggctt ccatttttac atgtgtatat aaccaatctt ttcctatctt 11400tgcttatatt ggtgtaactt tattttaata acatgtcaat gctatactgt taagagaagg 11460tctgaggaag attaagaaaa aggcctcgtg ttcacttggt tgccgtcaag tatcctgtgg 11520tttttttcta cctaacttcc tcatgccata tggctaccca gcatacccag tacccggatg 11580cacgtttatc ctcacctata gtcctggatc aatgtgattt ggtaactcga gcatgtgggt 11640tatattcatc ttattctcta aatcctcaac taaggcaatg taaattacca aaacatatat 11700atcgacttaa gttcgacaca atagtatcca aattcctaag tgatacacct gtagcaacac 11760tgccgataga ctatttagta ccaattctcc tgcgttccct aacggggcac ggtgataggc 11820cattgacccc gacttgcaat caattccttg atgaaattat taattacact cttcatgatg 11880cagcctttct tgattactat ctcaaggcaa caggtgcaca ggaccatttg acaaacattg 11940caactagaga gaagcttaaa aacgaaattc taaacaatga ttatgtccat caattgttct 12000tctggcatga cctttctatt ttggctcgac gtgggcgtct gaatcgcggg aacaaccgtt 12060caacctggtt tgttcatgat gaattcattg atattttagg atatggcgat tatatttttt 12120ggagaatacc tttatcatta ttaccagtta ctatagacgg ggtcccacac gcagcaactg 12180actggtatca accgactctt tttaaagaat ccatcctagg gcatagccaa atcctatctg 12240tgtcaacagc tgaaatacta attatgtgta aagatattat cacctgtagg tttaatacat 12300cactgattgc atccattgca aaattagagg atgtagatgt gtctgattat cctgacccga 12360gtgatattct taagatatac aatgctggag actatgtaat atctattctt ggctcagagg 12420gttataagat aataaagtac cttgaaccac tttgtttggc caaaatccaa ctttgctcta 12480aattcacaga aagaaaaggt cgtttcctca cacagatgcg tttatcagta ataaatgatc 12540ttcgggagtt gatttctaac cgcaggttaa aggactatca gcaagagaag attagagatt 12600ttcacaaaat attattacaa ttgcaattat ctcctcaaca gttttgtgaa ttattctctg 12660ttcaaaaaca ttgggggcat ccaattttac atagtgagaa agctatacaa aaagtaaaac 12720ggcatgcaac catccttaag gctctcagac ctaatgtcat ctttgagaca tattgtgtat 12780tcaagtacaa tattgccaag cactatttcg acagccaagg aacttggtac agtgtaatct 12840cagacaggaa tttaactcca ggactcaact ccgtcataaa acgtaatcac tttccttcac 12900tacccatgat taaggatctt ctatgggaat tctatcatct taatcaccct ccgttattct 12960ctacaaaggt gattagtgac ttaagtattt tcatcaaaga tagggccaca gctgttgaac 13020agacatgttg ggatgcagtc tttgaaccca atgtgctagg ttacaatcct ccaaacaaat 13080tctccactaa aagggtgccg gaacaatttc tagaacaaga ggatttttca atcgaaagtg 13140tcctgaatta tgcacaggaa ttacattatt tattaccaca gaataggaat ttttcctttt 13200ctctcaaaga aaaggaatta aatattggac gaacatttgg gaagctacca tatctcacac 13260ggaatgtcca aactttatgt gaggctctgt tagcagatgg actggccaag gccttcccca 13320gtaacatgat ggtagtaact gaacgtggac aaaaagagag ccttcttcat caggcatcat 13380ggcaccacac cagtgatgat tttggagaga atgctaccgt tcgagggagt agttttgtaa 13440ctgatttaga gaagtacaat cttgcatttc gctatgagtt cactgcacca tttattgagt 13500actgcaacca ttgctatggt gtgcgtaatg tctttaattg gatgcattat ttaatcccgc 13560agtgttacat gcatgtaagt gattattata atccgcctca caatgttaat cttagcaatc 13620gagaatatcc tcctgaaggc ccgagttcgt accgagggca cttaggaggc atagagggat 13680tacaacaaaa actgtggacg agtatatcct gtgcacaaat ctccttagtg gaaattaaaa 13740ctggttttaa gttacgatca gcggtcatgg gagacaatca gtgtataacc gtattgtctg 13800tttttccact tgaaacagac cctgaagagc aggagcaaag cgccgaagac aatgctgcaa 13860gagtagcagc aagtcttgca aaagtaacca gtgcatgtgg gatctttctt aaaccagatg 13920agacatttgt acactcaggt ttcatttatt tcggaaaaaa acaatatctc aatggtgtac 13980aattaccgca atcactcaaa acagcagcaa gaatggcacc actctctgat gctatattcg 14040atgatctaca aggaacactt gccagtattg gaactgcctt cgaacgtgct atatcggaaa 14100cgcgacatat cctcccatgt cgtattgtag cagctttcca tacgtatttc gccgttcgga 14160ttttacaata tcaccatctt ggatttaata aaggcatcga tttaggacag ttgtcactta 14220gtaaaccatt agactatggg actattactc taacattggc ggttccacaa gtccttgggg 14280gattgtcttt tctaaatcca gaaaagtgtt tttatcgaaa cttcggagat cctgtgactt 14340ctggactttt ccagctacgg gtgtacctag aaatggttaa catgaaagac ctattttgtc 14400cattaatatc gaaaaatcca ggaaattgta gtgccattga ttttgtctta aatccatccg 14460gattaaatgt tccaggatca caagacttga catccttttt gcgacaaatc gttaggcgta 14520gtattaccct aactgctaga aataagttaa ttaacactct cttccatgcc tctgctgatt 14580tggaagatga gatggtttgt aagtggctcc tttcatcaaa ccctgtcatg agtcgctttg 14640cagcggatat tttttccagg acaccgagtg gtaaacgtct ccaaatatta ggttatcttg 14700aagggaccag gactctattg gcctccaaaa tcataaacaa caacagtgag acacctgtac 14760ttgataagct gaggaagatc accctacaaa gatggaatct gtggttcagt tatttggacc 14820attgtgacca attactagca gatgctctac agaaaattag ttgcacggtg gatttggccc 14880agattttgcg tgagtataca tggtcacaca tcttagaggg tagatcattg atcggagcga 14940cattaccatg tatggtggag caattcaaag ttaagtggct aggacaatat gaaccttgtc 15000cagaatgtct caacaaaaaa ggctcaaatg cttatgtctc agttgcagtc aaagatcaag 15060tggtcagtgc ttggcctaat acttctcgaa taagttggac aatagggagt ggtgtcccct 15120atatagggtc aagaaccgag gataaaatcg gacagcctgc tatcaagccg cgatgccctt 15180catctgccct caaggaggct atagaattag catcaaggct cacttgggtt acacaaggag 15240gttctaatag tgaacaatta atccggcctt tcttggaagc gagagtcaac cttagtgtca 15300gtgaagtcct gcaaatgaca ccatcacatt attcaggaaa tattgtccat cgatataacg 15360accaatacag cccgcactca tttatggcga atcgcatgag caatactgcg acccgtctca 15420tagtgtcaac taatacactt ggagaatttt caggtggagg gcaggccgcc agggatagca 15480atataatttt ccagaatgtt ataaatttag cagttgccct ttatgatatt agattccgga 15540atacaaacac ctctgatata aggcataata gggctcatct tcacctgaca gagtgctgta 15600ctaaagaggt cccggcccag tatttgacat atacaagtgc acttaatctg gatttgagcc 15660gttatcgtgg taatgaacta atatatgact caaatccact gaagggagga ttgaactgca 15720atttaacaat agatagtcct ttagtgaagg gtcctaggct taacatgatt gaagatgatc 15780ttctccgctt tccacacctt tctggatggg agttagcgaa aacggtggta caatccatca 15840tctcagacaa tagcaactca tcaacagatc caatcagtag cggagaaaca cgctctttca 15900caactcattt tctcacttac cctcagattg gccttcttta cagtttcgga gcagtattat 15960gcttttatct aggcaatact atcctatgga ctaaaaaact tgattacgac cagtttctat 16020attatttgca taaccagctg cacaacttac ctcatcgagc actccgtgtt tttaaaccaa 16080catttaagca tgccagtgtg atgtcccgat taatggaaat tgattctaac ttctcaattt 16140atattggcgg gacatctgga gatcgagggc tgtctgatgc tgctcgactg tttcttcgga 16200cagcaatcgc gagtttttta caatttctta aaagctggat catcgatcgc caaaagacaa 16260ttcctttatg gatagtatat ccgcttgaag gtcaacagcc ggaatccatc aatgaatttc 16320tacataaaat tttgggtctg ctcaaacaag gccccaaaag tattccaaag gaggtcagca 16380tccaaaatga tggacatttg gatttggcag aaaataatta tgtttacaat agtaagagca 16440ctgctagtaa tttcttccat gcatccttag cttactggag aagtaggaaa tctcggaaaa 16500ctcaagacca taatgatttc tcaagagggg atggaacact tacagaaccc gtgcgtaagt 16560tttcaagcaa tcatcagtca gatgcaaagt actacaatgt gacatgtgga aagtcaccga 16620agccgcaaga acgcaaagac ttctcgcaat acagactcag caataacggg caaacaatga 16680gtaatcatcg taagaaaggg aagttccaca agtggaatcc ctgcaaaatg ttaatggaga 16740gtcaaagggg aactgttcta acagagggtg actactttca aaacaatact ccaccaacag 16800atgatgtatc aagtcctcac cgactcattc taccattttt taaattggga aatcacaacc 16860atgcacatga tcaagatgcc caagaattga tgaatcaaaa tattaagcag tacctacatc 16920agctaaggtc tatgttggac accactatat attgtagatt cacagggata gtctcatcca 16980tgcattacaa attggacgaa gttcttctag aatacaatag tttcgattca gctatcacat 17040tagctgaagg tgaggggtca ggggctctat tacttttgca aaaatatagt acaaggttat 17100tatttttgaa cacattggca acagaacaca gtatagaatc agaagttgta tcaggttttt 17160ctactccgag aatgttgtta ccaataatgc aaaaggttca tgaaggacaa gtcactgtta 17220tcttaaataa ttcatcaagt cacataactg acataactag ctcaatgtgg ctaagtagta 17280atcaaaaata taatctacct tgtcaagttg aaatcattat gatggatgct gaaacaacag 17340agaacttaaa caggtcccaa ctctaccgag cagtatataa cttaatactt gatcacattg 17400atccgcagta tctcaaggtg gtggtactca aagtatttct gagtgatata gaaggaatat 17460tatggattaa tgattacttg gctccattat tcggggctgg ttacttgatt aaaccgatta 17520catcaagtgc ccggtcaagt gaatggtacc tttgcttatc aaatttgata tctactaaca 17580ggagatcggc ccatcagact cacaaggcat gtcttggtgt tatcagagat gctttgcaag 17640cacaagtcca gcgaggcgtg tactggttga gtcacatcgc acagtatgct acaaagaatc 17700tccattgtga atacataggc cttggtttcc catctctaga aaaggtccta tatcacaggt 17760ataatctagt tgatactgga ctcggtccat tgtcgtcagt tattagacat ttaactaacc 17820tccaggcaga gatacgagac ttagtattag attataacct gatgagggag agtcgcactc 17880aaacgtacca ttttattaag actgcaaaag gcagaatcac aaagttagtc aatgactttc 17940tgaagttttc tttaattgtc caggcactca aaaataattc ttcttggtat actgagctta 18000aaaaattacc tgaggttatt aatgtgtgta atcgatttta tcatactcac aattgcgaat 18060gtcaggaaaa attctttgtc cagacgcttt atttacaacg cctacgcgat gcagaaatca 18120agctaattga acgccttacc gggttaatgc gattttatcc agaagggtta atatattcca 18180atcacacata ggtactaaat catcatagta tgaggaataa gataatgata attcctgacg 18240acagttttag ttccgattct aagtatatcg gaagagagta tgccaatctt aattgttaga 18300ggtaacaagc tattagttat tacttattga taagaataca ctttatcata gcgtaacaca 18360tcataacttt ataacgattt tgcatttcta atcctagtat ttattagaat gtactaccag 18420agaaatgacc ccagttccta tctttaaata atgattgtgt gtattaaatt attagtttat 18480taggtttatg agttggttac acagtgagta ttagtaattg aggattatgt agataggtaa 18540tctaacactg aatcacccat ctgatgtcac catatccaaa tgttgtgcta gtcgcattta 18600aacatgctat cttcagttaa gtaacatagg ctgaaaatgc taagaagaga ttggagtaaa 18660agtataaaat aaatttaatt aaacttcaaa gtgattaaat gataatgatc ttgggaactc 18720gatatgacct caagtcaaaa ataatgtcaa tataattgtt tagtaatatg agtgataatg 18780taaattttga taactaacta gctttagtag ttaagatcaa atgcaaacat

tataagaatg 18840ttaagcgcac acaaaaacat tataaaaaac caattttttc ctttttgtgt gtcca 188956288PRTReston ebolavirus 6Met Met Glu His Ser Arg Glu Arg Gly Arg Ser Ser Asn Met Arg His1 5 10 15 Asn Ser Arg Glu Pro Tyr Glu Asn Pro Ser Arg Ser Arg Ser Leu Ser 20 25 30 Arg Asp Pro Asn Gln Val Asp Arg Arg Gln Pro Arg Ser Ala Ser Gln 35 40 45 Ile Arg Val Pro Asn Leu Phe His Arg Lys Lys Thr Asp Ala Leu Ile 50 55 60 Val Pro Pro Thr Pro Lys Asp Ile Cys Pro Thr Leu Lys Lys Gly Phe65 70 75 80 Leu Cys Asp Ser Lys Phe Cys Lys Lys Asp His Gln Leu Asp Ser Leu 85 90 95 Asn Asp His Glu Leu Leu Leu Leu Ile Ala Arg Arg Thr Cys Gly Ile 100 105 110 Ile Glu Ser Asn Ser Gln Ile Thr Ser Pro Lys Asp Met Arg Leu Ala 115 120 125 Asn Pro Thr Ala Glu Asp Phe Ser His Gly Asn Ser Pro Lys Leu Thr 130 135 140 Leu Ala Val Leu Leu Gln Ile Ala Glu His Trp Ala Thr Arg Asp Leu145 150 155 160 Arg Gln Ile Glu Asp Ser Lys Leu Arg Ala Leu Leu Thr Leu Cys Ala 165 170 175 Val Leu Thr Arg Lys Phe Ser Lys Ser Gln Leu Gly Leu Leu Cys Glu 180 185 190 Thr His Leu Arg His Glu Gly Leu Gly Gln Asp Gln Ala Asp Ser Val 195 200 205 Leu Glu Val Tyr Gln Arg Leu His Ser Asp Lys Gly Gly Asn Phe Glu 210 215 220 Ala Ala Leu Trp Gln Gln Trp Asp Arg Gln Ser Leu Ile Met Phe Ile225 230 235 240 Ser Ala Phe Leu Asn Ile Ala Leu Gln Ile Pro Cys Glu Ser Ser Ser 245 250 255 Val Val Val Ser Gly Leu Ala Thr Leu Tyr Pro Ala Gln Asp Asn Ser 260 265 270 Thr Pro Ser Glu Ala Thr Asn Asp Thr Thr Trp Ser Ser Thr Val Glu 275 280 285 718959DNAZaire ebolavirus 7cggacacaca aaaagaaaga agaattttta ggatcttttg tgtgcgaata actatgagga 60agattaataa ttttcctctc attgaaattt atatcggaat ttaaattgaa attgttactg 120taatcacacc tggtttgttt cagagccaca tcacaaagat agagaacaac ctaggtctcc 180gaagggagca agggcatcag tgtgctcagt tgaaaatccc ttgtcaacac ctaggtctta 240tcacatcaca agttccacct cagactctgc agggtgatcc aacaacctta atagaaacat 300tattgttaaa ggacagcatt agttcacagt caaacaagca agattgagaa ttaaccttgg 360ttttgaactt gaacacttag gggattgaag attcaacaac cctaaagctt ggggtaaaac 420attggaaata gttaaaagac aaattgctcg gaatcacaaa attccgagta tggattctcg 480tcctcagaaa atctggatgg cgccgagtct cactgaatct gacatggatt accacaagat 540cttgacagca ggtctgtccg ttcaacaggg gattgttcgg caaagagtca tcccagtgta 600tcaagtaaac aatcttgaag aaatttgcca acttatcata caggcctttg aagcaggtgt 660tgattttcaa gagagtgcgg acagtttcct tctcatgctt tgtcttcatc atgcgtacca 720gggagattac aaacttttct tggaaagtgg cgcagtcaag tatttggaag ggcacgggtt 780ccgttttgaa gtcaagaagc gtgatggagt gaagcgcctt gaggaattgc tgccagcagt 840atctagtgga aaaaacatta agagaacact tgctgccatg ccggaagagg agacaactga 900agctaatgcc ggtcagtttc tctcctttgc aagtctattc cttccgaaat tggtagtagg 960agaaaaggct tgccttgaga aggttcaaag gcaaattcaa gtacatgcag agcaaggact 1020gatacaatat ccaacagctt ggcaatcagt aggacacatg atggtgattt tccgtttgat 1080gcgaacaaat tttctgatca aatttctcct aatacaccaa gggatgcaca tggttgccgg 1140gcatgatgcc aacgatgctg tgatttcaaa ttcagtggct caagctcgtt tttcaggctt 1200attgattgtc aaaacagtac ttgatcatat cctacaaaag acagaacgag gagttcgtct 1260ccatcctctt gcaaggaccg ccaaggtaaa aaatgaggtg aactccttta aggctgcact 1320cagctccctg gccaagcatg gagagtatgc tcctttcgcc cgacttttga acctttctgg 1380agtaaataat cttgagcatg gtcttttccc tcaactatcg gcaattgcac tcggagtcgc 1440cacagcacac gggagtaccc tcgcaggagt aaatgttgga gaacagtatc aacaactcag 1500agaggctgcc actgaggctg agaagcaact ccaacaatat gcagagtctc gcgaacttga 1560ccatcttgga cttgatgatc aggaaaagaa aattcttatg aacttccatc agaaaaagaa 1620cgaaatcagc ttccagcaaa caaacgctat ggtaactcta agaaaagagc gcctggccaa 1680gctgacagaa gctatcactg ctgcgtcact gcccaaaaca agtggacatt acgatgatga 1740tgacgacatt ccctttccag gacccatcaa tgatgacgac aatcctggcc atcaagatga 1800tgatccgact gactcacagg atacgaccat tcccgatgtg gtggttgatc ccgatgatgg 1860aagctacggc gaataccaga gttactcgga aaacggcatg aatgcaccag atgacttggt 1920cctattcgat ctagacgagg acgacgagga cactaagcca gtgcctaata gatcgaccaa 1980gggtggacaa cagaagaaca gtcaaaaggg ccagcatata gagggcagac agacacaatc 2040caggccaatt caaaatgtcc caggccctca cagaacaatc caccacgcca gtgcgccact 2100cacggacaat gacagaagaa atgaaccctc cggctcaacc agccctcgca tgctgacacc 2160aattaacgaa gaggcagacc cactggacga tgccgacgac gagacgtcta gccttccgcc 2220cttggagtca gatgatgaag agcaggacag ggacggaact tccaaccgca cacccactgt 2280cgccccaccg gctcccgtat acagagatca ctctgaaaag aaagaactcc cgcaagacga 2340gcaacaagat caggaccaca ctcaagaggc caggaaccag gacagtgaca acacccagtc 2400agaacactct tttgaggaga tgtatcgcca cattctaaga tcacaggggc catttgatgc 2460tgttttgtat tatcatatga tgaaggatga gcctgtagtt ttcagtacca gtgatggcaa 2520agagtacacg tatccagact cccttgaaga ggaatatcca ccatggctca ctgaaaaaga 2580ggctatgaat gaagagaata gatttgttac attggatggt caacaatttt attggccggt 2640gatgaatcac aagaataaat tcatggcaat cctgcaacat catcagtgaa tgagcatgga 2700acaatgggat gattcaaccg acaaatagct aacattaagt agtcaaggaa cgaaaacagg 2760aagaattttt gatgtctaag gtgtgaatta ttatcacaat aaaagtgatt cttatttttg 2820aatttaaagc tagcttatta ttactagccg tttttcaaag ttcaatttga gtcttaatgc 2880aaataggcgt taagccacag ttatagccat aattgtaact caatattcta actagcgatt 2940tatctaaatt aaattacatt atgcttttat aacttaccta ctagcctgcc caacatttac 3000acgatcgttt tataattaag aaaaaactaa tgatgaagat taaaaccttc atcatcctta 3060cgtcaattga attctctagc actcgaagct tattgtcttc aatgtaaaag aaaagctggt 3120ctaacaagat gacaactaga acaaagggca ggggccatac tgcggccacg actcaaaacg 3180acagaatgcc aggccctgag ctttcgggct ggatctctga gcagctaatg accggaagaa 3240ttcctgtaag cgacatcttc tgtgatattg agaacaatcc aggattatgc tacgcatccc 3300aaatgcaaca aacgaagcca aacccgaaga cgcgcaacag tcaaacccaa acggacccaa 3360tttgcaatca tagttttgag gaggtagtac aaacattggc ttcattggct actgttgtgc 3420aacaacaaac catcgcatca gaatcattag aacaacgcat tacgagtctt gagaatggtc 3480taaagccagt ttatgatatg gcaaaaacaa tctcctcatt gaacagggtt tgtgctgaga 3540tggttgcaaa atatgatctt ctggtgatga caaccggtcg ggcaacagca accgctgcgg 3600caactgaggc ttattgggcc gaacatggtc aaccaccacc tggaccatca ctttatgaag 3660aaagtgcgat tcggggtaag attgaatcta gagatgagac cgtccctcaa agtgttaggg 3720aggcattcaa caatctaaac agtaccactt cactaactga ggaaaatttt gggaaacctg 3780acatttcggc aaaggatttg agaaacatta tgtatgatca cttgcctggt tttggaactg 3840ctttccacca attagtacaa gtgatttgta aattgggaaa agatagcaac tcattggaca 3900tcattcatgc tgagttccag gccagcctgg ctgaaggaga ctctcctcaa tgtgccctaa 3960ttcaaattac aaaaagagtt ccaatcttcc aagatgctgc tccacctgtc atccacatcc 4020gctctcgagg tgacattccc cgagcttgcc agaaaagctt gcgtccagtc ccaccatcgc 4080ccaagattga tcgaggttgg gtatgtgttt ttcagcttca agatggtaaa acacttggac 4140tcaaaatttg agccaatctc ccttccctcc gaaagaggcg aataatagca gaggcttcaa 4200ctgctgaact atagggtacg ttacattaat gatacacttg tgagtatcag ccctggataa 4260tataagtcaa ttaaacgacc aagataaaat tgttcatatc tcgctagcag cttaaaatat 4320aaatgtaata ggagctatat ctctgacagt attataatca attgttatta agtaacccaa 4380accaaaagtg atgaagatta agaaaaacct acctcggctg agagagtgtt ttttcattaa 4440ccttcatctt gtaaacgttg agcaaaattg ttaaaaatat gaggcgggtt atattgccta 4500ctgctcctcc tgaatatatg gaggccatat accctgtcag gtcaaattca acaattgcta 4560gaggtggcaa cagcaataca ggcttcctga caccggagtc agtcaatggg gacactccat 4620cgaatccact caggccaatt gccgatgaca ccatcgacca tgccagccac acaccaggca 4680gtgtgtcatc agcattcatc cttgaagcta tggtgaatgt catatcgggc cccaaagtgc 4740taatgaagca aattccaatt tggcttcctc taggtgtcgc tgatcaaaag acctacagct 4800ttgactcaac tacggccgcc atcatgcttg cttcatacac tatcacccat ttcggcaagg 4860caaccaatcc acttgtcaga gtcaatcggc tgggtcctgg aatcccggat catcccctca 4920ggctcctgcg aattggaaac caggctttcc tccaggagtt cgttcttccg ccagtccaac 4980taccccagta tttcaccttt gatttgacag cactcaaact gatcacccaa ccactgcctg 5040ctgcaacatg gaccgatgac actccaacag gatcaaatgg agcgttgcgt ccaggaattt 5100catttcatcc aaaacttcgc cccattcttt tacccaacaa aagtgggaag aaggggaaca 5160gtgccgatct aacatctccg gagaaaatcc aagcaataat gacttcactc caggacttta 5220agatcgttcc aattgatcca accaaaaata tcatgggaat cgaagtgcca gaaactctgg 5280tccacaagct gaccggtaag aaggtgactt ctaaaaatgg acaaccaatc atccctgttc 5340ttttgccaaa gtacattggg ttggacccgg tggctccagg agacctcacc atggtaatca 5400cacaggattg tgacacgtgt cattctcctg caagtcttcc agctgtgatt gagaagtaat 5460tgcaataatt gactcagatc cagttttata gaatcttctc agggatagtg ataacatcta 5520tttagtaatc cgtccattag aggagacact tttaattgat caatatacta aaggtgcttt 5580acaccattgt cttttttctc tcctaaatgt agaacttaac aaaagactca taatatactt 5640gtttttaaag gattgattga tgaaagatca taactaataa cattacaaat aatcctacta 5700taatcaatac ggtgattcaa atgttaatct ttctcattgc acatactttt tgcccttatc 5760ctcaaattgc ctgcatgctt acatctgagg atagccagtg tgacttggat tggaaatgtg 5820gagaaaaaat cgggacccat ttctaggttg ttcacaatcc aagtacagac attgcccttc 5880taattaagaa aaaatcggcg atgaagatta agccgacagt gagcgtaatc ttcatctctc 5940ttagattatt tgttttccag agtaggggtc gtcaggtcct tttcaatcgt gtaaccaaaa 6000taaactccac tagaaggata ttgtggggca acaacacaat gggcgttaca ggaatattgc 6060agttacctcg tgatcgattc aagaggacat cattctttct ttgggtaatt atccttttcc 6120aaagaacatt ttccatccca cttggagtca tccacaatag cacattacag gttagtgatg 6180tcgacaaact agtttgtcgt gacaaactgt catccacaaa tcaattgaga tcagttggac 6240tgaatctcga agggaatgga gtggcaactg acgtgccatc tgcaactaaa agatggggct 6300tcaggtccgg tgtcccacca aaggtggtca attatgaagc tggtgaatgg gctgaaaact 6360gctacaatct tgaaatcaaa aaacctgacg ggagtgagtg tctaccagca gcgccagacg 6420ggattcgggg cttcccccgg tgccggtatg tgcacaaagt atcaggaacg ggaccgtgtg 6480ccggagactt tgccttccat aaagagggtg ctttcttcct gtatgatcga cttgcttcca 6540cagttatcta ccgaggaacg actttcgctg aaggtgtcgt tgcatttctg atactgcccc 6600aagctaagaa ggacttcttc agctcacacc ccttgagaga gccggtcaat gcaacggagg 6660acccgtctag tggctactat tctaccacaa ttagatatca ggctaccggt tttggaacca 6720atgagacaga gtacttgttc gaggttgaca atttgaccta cgtccaactt gaatcaagat 6780tcacaccaca gtttctgctc cagctgaatg agacaatata tacaagtggg aaaaggagca 6840ataccacggg aaaactaatt tggaaggtca accccgaaat tgatacaaca atcggggagt 6900gggccttctg ggaaactaaa aaaacctcac tagaaaaatt cgcagtgaag agttgtcttt 6960cacagttgta tcaaacggag ccaaaaacat cagtggtcag agtccggcgc gaacttcttc 7020cgacccaggg accaacacaa caactgaaga ccacaaaatc atggcttcag aaaattcctc 7080tgcaatggtt caagtgcaca gtcaaggaag ggaagctgca gtgtcgcatc taacaaccct 7140tgccacaatc tccacgagtc cccaatccct cacaaccaaa ccaggtccgg acaacagcac 7200ccataataca cccgtgtata aacttgacat ctctgaggca actcaagttg aacaacatca 7260ccgcagaaca gacaacgaca gcacagcctc cgacactccc tctgccacga ccgcagccgg 7320acccccaaaa gcagagaaca ccaacacgag caagagcact gacttcctgg accccgccac 7380cacaacaagt ccccaaaacc acagcgagac cgctggcaac aacaacactc atcaccaaga 7440taccggagaa gagagtgcca gcagcgggaa gctaggctta attaccaata ctattgctgg 7500agtcgcagga ctgatcacag gcgggagaag aactcgaaga gaagcaattg tcaatgctca 7560acccaaatgc aaccctaatt tacattactg gactactcag gatgaaggtg ctgcaatcgg 7620actggcctgg ataccatatt tcgggccagc agccgaggga atttacacag aggggctaat 7680gcacaatcaa gatggtttaa tctgtgggtt gagacagctg gccaacgaga cgactcaagc 7740tcttcaactg ttcctgagag ccacaactga gctacgcacc ttttcaatcc tcaaccgtaa 7800ggcaattgat ttcttgctgc agcgatgggg cggcacatgc cacattctgg gaccggactg 7860ctgtatcgaa ccacatgatt ggaccaagaa cataacagac aaaattgatc agattattca 7920tgattttgtt gataaaaccc ttccggacca gggggacaat gacaattggt ggacaggatg 7980gagacaatgg ataccggcag gtattggagt tacaggcgtt ataattgcag ttatcgcttt 8040attctgtata tgcaaatttg tcttttagtt tttcttcaga ttgcttcatg gaaaagctca 8100gcctcaaatc aatgaaacca ggatttaatt atatggatta cttgaatcta agattacttg 8160acaaatgata atataataca ctggagcttt aaacatagcc aatgtgattc taactccttt 8220aaactcacag ttaatcataa acaaggtttg acatcaatct agttatctct ttgagaatga 8280taaacttgat gaagattaag aaaaaggtaa tctttcgatt atctttaatc ttcatccttg 8340attctacaat catgacagtt gtctttagtg acaagggaaa gaagcctttt tattaagttg 8400taataatcag atctgcgaac cggtagagtt tagttgcaac ctaacacaca taaagcattg 8460gtcaaaaagt caatagaaat ttaaacagtg agtggagaca acttttaaat ggaagcttca 8520tatgagagag gacgcccacg agctgccaga cagcattcaa gggatggaca cgaccaccat 8580gttcgagcac gatcatcatc cagagagaat tatcgaggtg agtaccgtca atcaaggagc 8640gcctcacaag tgcgcgttcc tactgtattt cataagaaga gagttgaacc attaacagtt 8700cctccagcac ctaaagacat atgtccgacc ttgaaaaaag gatttttgtg tgacagtagt 8760ttttgcaaaa aagatcacca gttggagagt ttaactgata gggaattact cctactaatc 8820gcccgtaaga cttgtggatc agtagaacaa caattaaata taactgcacc caaggactcg 8880cgcttagcaa atccaacggc tgatgatttc cagcaagagg aaggtccaaa aattaccttg 8940ttgacactga tcaagacggc agaacactgg gcgagacaag acatcagaac catagaggat 9000tcaaaattaa gagcattgtt gactctatgt gctgtgatga cgaggaaatt ctcaaaatcc 9060cagctgagtc ttttatgtga gacacaccta aggcgcgagg ggcttgggca agatcaggca 9120gaacccgttc tcgaagtata tcaacgatta cacagtgata aaggaggcag ttttgaagct 9180gcactatggc aacaatggga ccgacaatcc ctaattatgt ttatcactgc attcttgaat 9240attgctctcc agttaccgtg tgaaagttct gctgtcgttg tttcagggtt aagaacattg 9300gttcctcaat cagataatga ggaagcttca accaacccgg ggacatgctc atggtctgat 9360gagggtaccc cttaataagg ctgactaaaa cactatataa ccttctactt gatcacaata 9420ctccgtatac ctatcatcat atatttaatc aagacgatat cctttaaaac ttattcagta 9480ctataatcac tctcgtttca aattaataag atgtgcatga ttgccctaat atatgaagag 9540gtatgataca accctaacag tgatcaaaga aaatcataat ctcgtatcgc tcgtaatata 9600acctgccaag catacctctt gcacaaagtg attcttgtac acaaataatg ttttactcta 9660caggaggtag caacgatcca tcccatcaaa aaataagtat ttcatgactt actaatgatc 9720tcttaaaata ttaagaaaaa ctgacggaac ataaattctt tatgcttcaa gctgtggagg 9780aggtgtttgg tattggctat tgttatatta caatcaataa caagcttgta aaaatattgt 9840tcttgtttca agaggtagat tgtgaccgga aatgctaaac taatgatgaa gattaatgcg 9900gaggtctgat aagaataaac cttattattc agattaggcc ccaagaggca ttcttcatct 9960ccttttagca aagtactatt tcagggtagt ccaattagtg gcacgtcttt tagctgtata 10020tcagtcgccc ctgagatacg ccacaaaagt gtctctaagc taaattggtc tgtacacatc 10080ccatacattg tattaggggc aataatatct aattgaactt agccgtttaa aatttagtgc 10140ataaatctgg gctaacacca ccaggtcaac tccattggct gaaaagaagc ttacctacaa 10200cgaacatcac tttgagcgcc ctcacaatta aaaaatagga acgtcgttcc aacaatcgag 10260cgcaaggttt caaggttgaa ctgagagtgt ctagacaaca aaatattgat actccagaca 10320ccaagcaaga cctgagaaaa aaccatggct aaagctacgg gacgatacaa tctaatatcg 10380cccaaaaagg acctggagaa aggggttgtc ttaagcgacc tctgtaactt cttagttagc 10440caaactattc aggggtggaa ggtttattgg gctggtattg agtttgatgt gactcacaaa 10500ggaatggccc tattgcatag actgaaaact aatgactttg cccctgcatg gtcaatgaca 10560aggaatctct ttcctcattt atttcaaaat ccgaattcca caattgaatc accgctgtgg 10620gcattgagag tcatccttgc agcagggata caggaccagc tgattgacca gtctttgatt 10680gaacccttag caggagccct tggtctgatc tctgattggc tgctaacaac caacactaac 10740catttcaaca tgcgaacaca acgtgtcaag gaacaattga gcctaaaaat gctgtcgttg 10800attcgatcca atattctcaa gtttattaac aaattggatg ctctacatgt cgtgaactac 10860aacggattgt tgagcagtat tgaaattgga actcaaaatc atacaatcat cataactcga 10920actaacatgg gttttctggt ggagctccaa gaacccgaca aatcggcaat gaaccgcatg 10980aagcctgggc cggcgaaatt ttccctcctt catgagtcca cactgaaagc atttacacaa 11040ggatcctcga cacgaatgca aagtttgatt cttgaattta atagctctct tgctatctaa 11100ctaaggtaga atacttcata ttgagctaac tcatatatgc tgactcaata gttatcttga 11160catctctgct ttcataatca gatatataag cataataaat aaatactcat atttcttgat 11220aatttgttta accacagata aatcctcact gtaagccagc ttccaagttg acacccttac 11280aaaaaccagg actcagaatc cctcaaacaa gagattccaa gacaacatca tagaattgct 11340ttattatatg aataagcatt ttatcaccag aaatcctata tactaaatgg ttaattgtaa 11400ctgaacccgc aggtcacatg tgttaggttt cacagattct atatattact aactctatac 11460tcgtaattaa cattagataa gtagattaag aaaaaagcct gaggaagatt aagaaaaact 11520gcttattggg tctttccgtg ttttagatga agcagttgaa attcttcctc ttgatattaa 11580atggctacac aacataccca atacccagac gctaggttat catcaccaat tgtattggac 11640caatgtgacc tagtcactag agcttgcggg ttatattcat catactccct taatccgcaa 11700ctacgcaact gtaaactccc gaaacatatc taccgtttga aatacgatgt aactgttacc 11760aagttcttga gtgatgtacc agtggcgaca ttgcccatag atttcatagt cccagttctt 11820ctcaaggcac tgtcaggcaa tggattctgt cctgttgagc cgcggtgcca acagttctta 11880gatgaaatca ttaagtacac aatgcaagat gctctcttct tgaaatatta tctcaaaaat 11940gtgggtgctc aagaagactg tgttgatgaa cactttcaag agaaaatctt atcttcaatt 12000cagggcaatg aatttttaca tcaaatgttt ttctggtatg atctggctat tttaactcga 12060aggggtagat taaatcgagg aaactctaga tcaacatggt ttgttcatga tgatttaata 12120gacatcttag gctatgggga ctatgttttt tggaagatcc caatttcaat gttaccactg 12180aacacacaag gaatccccca tgctgctatg gactggtatc aggcatcagt attcaaagaa 12240gcggttcaag ggcatacaca cattgtttct gtttctactg ccgacgtctt gataatgtgc 12300aaagatttaa ttacatgtcg attcaacaca actctaatct caaaaatagc agagattgag 12360gatccagttt gttctgatta tcccaatttt aagattgtgt ctatgcttta ccagagcgga 12420gattacttac tctccatatt agggtctgat gggtataaaa ttattaagtt cctcgaacca 12480ttgtgcttgg ccaaaattca attatgctca aagtacactg agaggaaggg ccgattctta 12540acacaaatgc atttagctgt aaatcacacc ctagaagaaa ttacagaaat gcgtgcacta 12600aagccttcac aggctcaaaa gatccgtgaa ttccatagaa cattgataag gctggagatg 12660acgccacaac aactttgtga gctattttcc attcaaaaac actgggggca tcctgtgcta 12720catagtgaaa cagcaatcca aaaagttaaa aaacatgcta cggtgctaaa agcattacgc 12780cctatagtga ttttcgagac atactgtgtt tttaaatata gtattgccaa acattatttt 12840gatagtcaag gatcttggta cagtgttact tcagatagga atctaacacc gggtcttaat 12900tcttatatca aaagaaatca attccctccg ttgccaatga ttaaagaact actatgggaa 12960ttttaccacc ttgaccaccc tccacttttc tcaaccaaaa ttattagtga cttaagtatt 13020tttataaaag acagagctac cgcagtagaa aggacatgct

gggatgcagt attcgagcct 13080aatgttctag gatataatcc acctcacaaa tttagtacta aacgtgtacc ggaacaattt 13140ttagagcaag aaaacttttc tattgagaat gttctttcct acgcacaaaa actcgagtat 13200ctactaccac aatatcggaa cttttctttc tcattgaaag agaaagagtt gaatgtaggt 13260agaaccttcg gaaaattgcc ttatccgact cgcaatgttc aaacactttg tgaagctctg 13320ttagctgatg gtcttgctaa agcatttcct agcaatatga tggtagttac ggaacgtgag 13380caaaaagaaa gcttattgca tcaagcatca tggcaccaca caagtgatga ttttggtgaa 13440catgccacag ttagagggag tagctttgta actgatttag agaaatacaa tcttgcattt 13500agatatgagt ttacagcacc ttttatagaa tattgcaacc gttgctatgg tgttaagaat 13560gtttttaatt ggatgcatta tacaatccca cagtgttata tgcatgtcag tgattattat 13620aatccaccac ataacctcac actggagaat cgagacaacc cccccgaagg gcctagttca 13680tacaggggtc atatgggagg gattgaagga ctgcaacaaa aactctggac aagtatttca 13740tgtgctcaaa tttctttagt tgaaattaag actggtttta agttacgctc agctgtgatg 13800ggtgacaatc agtgcattac tgttttatca gtcttcccct tagagactga cgcagacgag 13860caggaacaga gcgccgaaga caatgcagcg agggtggccg ccagcctagc aaaagttaca 13920agtgcctgtg gaatcttttt aaaacctgat gaaacatttg tacattcagg ttttatctat 13980tttggaaaaa aacaatattt gaatggggtc caattgcctc agtcccttaa aacggctaca 14040agaatggcac cattgtctga tgcaattttt gatgatcttc aagggaccct ggctagtata 14100ggcactgctt ttgagcgatc catctctgag acacgacata tctttccttg caggataacc 14160gcagctttcc atacgttttt ttcggtgaga atcttgcaat atcatcatct cgggttcaat 14220aaaggttttg accttggaca gttaacactc ggcaaacctc tggatttcgg aacaatatca 14280ttggcactag cggtaccgca ggtgcttgga gggttatcct tcttgaatcc tgagaaatgt 14340ttctaccgga atctaggaga tccagttacc tcaggcttat tccagttaaa aacttatctc 14400cgaatgattg agatggatga tttattctta cctttaattg cgaagaaccc tgggaactgc 14460actgccattg actttgtgct aaatcctagc ggattaaatg tccctgggtc gcaagactta 14520acttcatttc tgcgccagat tgtacgcagg accatcaccc taagtgcgaa aaacaaactt 14580attaatacct tatttcatgc gtcagctgac ttcgaagacg aaatggtttg taaatggcta 14640ttatcatcaa ctcctgttat gagtcgtttt gcggccgata tcttttcacg cacgccgagc 14700gggaagcgat tgcaaattct aggatacctg gaaggaacac gcacattatt agcctctaag 14760atcatcaaca ataatacaga gacaccggtt ttggacagac tgaggaaaat aacattgcaa 14820aggtggagcc tatggtttag ttatcttgat cattgtgata atatcctggc ggaggcttta 14880acccaaataa cttgcacagt tgatttagca cagattctga gggaatattc atgggctcat 14940attttagagg gaagacctct tattggagcc acactcccat gtatgattga gcaattcaaa 15000gtgttttggc tgaaacccta cgaacaatgt ccgcagtgtt caaatgcaaa gcaaccaggt 15060gggaaaccat tcgtgtcagt ggcagtcaag aaacatattg ttagtgcatg gccgaacgca 15120tcccgaataa gctggactat cggggatgga atcccataca ttggatcaag gacagaagat 15180aagataggac aacctgctat taaaccaaaa tgtccttccg cagccttaag agaggccatt 15240gaattggcgt cccgtttaac atgggtaact caaggcagtt cgaacagtga cttgctaata 15300aaaccatttt tggaagcacg agtaaattta agtgttcaag aaatacttca aatgacccct 15360tcacattact caggaaatat tgttcacagg tacaacgatc aatacagtcc tcattctttc 15420atggccaatc gtatgagtaa ttcagcaacg cgattgattg tttctacaaa cactttaggt 15480gagttttcag gaggtggcca gtctgcacgc gacagcaata ttattttcca gaatgttata 15540aattatgcag ttgcactgtt cgatattaaa tttagaaaca ctgaggctac agatatccaa 15600tataatcgtg ctcaccttca tctaactaag tgttgcaccc gggaagtacc agctcagtat 15660ttaacataca catctacatt ggatttagat ttaacaagat accgagaaaa cgaattgatt 15720tatgacagta atcctctaaa aggaggactc aattgcaata tctcattcga taatccattt 15780ttccaaggta aacggctgaa cattatagaa gatgatctta ttcgactgcc tcacttatct 15840ggatgggagc tagccaagac catcatgcaa tcaattattt cagatagcaa caattcatct 15900acagacccaa ttagcagtgg agaaacaaga tcattcacta cccatttctt aacttatccc 15960aagataggac ttctgtacag ttttggggcc tttgtaagtt attatcttgg caatacaatt 16020cttcggacta agaaattaac acttgacaat tttttatatt acttaactac tcaaattcat 16080aatctaccac atcgctcatt gcgaatactt aagccaacat tcaaacatgc aagcgttatg 16140tcacggttaa tgagtattga tcctcatttt tctatttaca taggcggtgc tgcaggtgac 16200agaggactct cagatgcggc caggttattt ttgagaacgt ccatttcatc ttttcttaca 16260tttgtaaaag aatggataat taatcgcgga acaattgtcc ctttatggat agtatatccg 16320ctagagggtc aaaacccaac acctgtgaat aattttctct atcagatcgt agaactgctg 16380gtgcatgatt catcaagaca acaggctttt aaaactacca taagtgatca tgtacatcct 16440cacgacaatc ttgtttacac atgtaagagt acagccagca atttcttcca tgcatcattg 16500gcgtactgga ggagcagaca cagaaacagc aaccgaaaat acttggcaag agactcttca 16560actggatcaa gcacaaacaa cagtgatggt catattgaga gaagtcaaga acaaaccacc 16620agagatccac atgatggcac tgaacggaat ctagtcctac aaatgagcca tgaaataaaa 16680agaacgacaa ttccacaaga aaacacgcac cagggtccgt cgttccagtc ctttctaagt 16740gactctgctt gtggtacagc aaatccaaaa ctaaatttcg atcgatcgag acacaatgtg 16800aaatttcagg atcataactc ggcatccaag agggaaggtc atcaaataat ctcacaccgt 16860ctagtcctac ctttctttac attatctcaa gggacacgcc aattaacgtc atccaatgag 16920tcacaaaccc aagacgagat atcaaagtac ttacggcaat tgagatccgt cattgatacc 16980acagtttatt gtagatttac cggtatagtc tcgtccatgc attacaaact tgatgaggtc 17040ctttgggaaa tagagagttt caagtcggct gtgacgctag cagagggaga aggtgctggt 17100gccttactat tgattcagaa ataccaagtt aagaccttat ttttcaacac gctagctact 17160gagtccagta tagagtcaga aatagtatca ggaatgacta ctcctaggat gcttctacct 17220gttatgtcaa aattccataa tgaccaaatt gagattattc ttaacaactc agcaagccaa 17280ataacagaca taacaaatcc tacttggttt aaagaccaaa gagcaaggct acctaagcaa 17340gtcgaggtta taaccatgga tgcagagaca acagagaata taaacagatc gaaattgtac 17400gaagctgtat ataaattgat cttacaccat attgatccta gcgtattgaa agcagtggtc 17460cttaaagtct ttctaagtga tactgagggt atgttatggc taaatgataa tttagccccg 17520ttttttgcca ctggttattt aattaagcca ataacgtcaa gtgctagatc tagtgagtgg 17580tatctttgtc tgacgaactt cttatcaact acacgtaaga tgccacacca aaaccatctc 17640agttgtaaac aggtaatact tacggcattg caactgcaaa ttcaacgaag cccatactgg 17700ctaagtcatt taactcagta tgctgactgt gagttacatt taagttatat ccgccttggt 17760tttccatcat tagagaaagt actataccac aggtataacc tcgtcgattc aaaaagaggt 17820ccactagtct ctatcactca gcacttagca catcttagag cagagattcg agaattaact 17880aatgattata atcaacagcg acaaagtcgg actcaaacat atcactttat tcgtactgca 17940aaaggacgaa tcacaaaact agtcaatgat tatttaaaat tctttcttat tgtgcaagca 18000ttaaaacata atgggacatg gcaagctgag tttaagaaat taccagagtt gattagtgtg 18060tgcaataggt tctaccatat tagagattgc aattgtgaag aacgtttctt agttcaaacc 18120ttatatttac atagaatgca ggattctgaa gttaagctta tcgaaaggct gacagggctt 18180ctgagtttat ttccggatgg tctctacagg tttgattgaa ttaccgtgca tagtatcctg 18240atacttgcaa aggttggtta ttaacataca gattataaaa aactcataaa ttgctctcat 18300acatcatatt gatctaatct caataaacaa ctatttaaat aacgaaagga gtccctatat 18360tatatactat atttagcctc tctccctgcg tgataatcaa aaaattcaca atgcagcatg 18420tgtgacatat tactgccgca atgaatttaa cgcaacataa taaactctgc actctttata 18480attaagcttt aacgaaaggt ctgggctcat attgttattg atataataat gttgtatcaa 18540tatcctgtca gatggaatag tgttttggtt gataacacaa cttcttaaaa caaaattgat 18600ctttaagatt aagtttttta taattatcat tactttaatt tgtcgtttta aaaacggtga 18660tagccttaat ctttgtgtaa aataagagat taggtgtaat aaccttaaca tttttgtcta 18720gtaagctact atttcataca gaatgataaa attaaaagaa aaggcaggac tgtaaaatca 18780gaaatacctt ctttacaata tagcagacta gataataatc ttcgtgttaa tgataattaa 18840gacattgacc acgctcatca gaaggctcgc cagaataaac gttgcaaaaa ggattcctgg 18900aaaaatggtc gcacacaaaa atttaaaaat aaatctattt cttctttttt gtgtgtcca 189598288PRTZaire ebolavirus 8Met Glu Ala Ser Tyr Glu Arg Gly Arg Pro Arg Ala Ala Arg Gln His1 5 10 15 Ser Arg Asp Gly His Asp His His Val Arg Ala Arg Ser Ser Ser Arg 20 25 30 Glu Asn Tyr Arg Gly Glu Tyr Arg Gln Ser Arg Ser Ala Ser Gln Val 35 40 45 Arg Val Pro Thr Val Phe His Lys Lys Arg Val Glu Pro Leu Thr Val 50 55 60 Pro Pro Ala Pro Lys Asp Ile Cys Pro Thr Leu Lys Lys Gly Phe Leu65 70 75 80 Cys Asp Ser Ser Phe Cys Lys Lys Asp His Gln Leu Glu Ser Leu Thr 85 90 95 Asp Arg Glu Leu Leu Leu Leu Ile Ala Arg Lys Thr Cys Gly Ser Val 100 105 110 Glu Gln Gln Leu Asn Ile Thr Ala Pro Lys Asp Ser Arg Leu Ala Asn 115 120 125 Pro Thr Ala Asp Asp Phe Gln Gln Glu Glu Gly Pro Lys Ile Thr Leu 130 135 140 Leu Thr Leu Ile Lys Thr Ala Glu His Trp Ala Arg Gln Asp Ile Arg145 150 155 160 Thr Ile Glu Asp Ser Lys Leu Arg Ala Leu Leu Thr Leu Cys Ala Val 165 170 175 Met Thr Arg Lys Phe Ser Lys Ser Gln Leu Ser Leu Leu Cys Glu Thr 180 185 190 His Leu Arg Arg Glu Gly Leu Gly Gln Asp Gln Ala Glu Pro Val Leu 195 200 205 Glu Val Tyr Gln Arg Leu His Ser Asp Lys Gly Gly Ser Phe Glu Ala 210 215 220 Ala Leu Trp Gln Gln Trp Asp Arg Gln Ser Leu Ile Met Phe Ile Thr225 230 235 240 Ala Phe Leu Asn Ile Ala Leu Gln Leu Pro Cys Glu Ser Ser Ala Val 245 250 255 Val Val Ser Gly Leu Arg Thr Leu Val Pro Gln Ser Asp Asn Glu Glu 260 265 270 Ala Ser Thr Asn Pro Gly Thr Cys Ser Trp Ser Asp Glu Gly Thr Pro 275 280 285 918891DNAReston ebolavirus 9cggacacaca aaaagaaaaa aggtttttta agactttttg tgtgcgagta actatgagga 60agattaacag ttttcctcag tttaagatat acactgaaat tgagattgag attctcctct 120ttgctattct gtaactttcc ctggttgtga caattgaatc agttttatct attaccaatt 180accatcaaca tggtatgtct agtgatcttg ggactcttct tcatctggtt tttcctagag 240ctctgaatcc attttgcgag aagttcatcc aaacgaccca gtgtctgaaa atacaaaagg 300ttcccctttc cgtcaagttt aaggggttgt tttgattgtg tgtagatttt ataatcctag 360agtgccaagg agttgcgtgt catcattgat tgggaagatc aaggaaacaa tttgttccaa 420taatatcgta catcttgact aagtcgaaca aggggaagtc gatatggatc gtgggaccag 480aagaatctgg gtgtcgcaaa atcaaggtga tactgattta gattatcata aaattttgac 540agctggcctt actgttcaac agggaattgt caggcagaaa ataatttctg tatatcttgt 600tgataacttg gaggctatgt gtcaattggt aatacaagcc tttgaggccg gaattgattt 660ccaagaaaat gccgacagct tccttctgat gctttgccta catcatgctt accaaggtga 720ctataaattg ttcttggaga gcaatgctgt acagtatttg gaaggtcatg gattcaaatt 780tgagctccgg aagaaggacg gtgtcaatcg gctcgaggaa ttgcttcctg ctgcaacgag 840tggaaaaaac atcaggcgta cgttggccgc actgcctgaa gaggagacta cagaagcaaa 900tgcagggcaa tttctctcat ttgcgagttt gtttcttccc aaactggttg tgggagagaa 960ggcttgcttg gaaaaagtcc agcgacaaat tcaggttcat gcagaacagg gtttaattca 1020atatcccact gcatggcaat cagttggaca catgatggta atcttcagat tgatgaggac 1080taatttcttg attaaatatt tactgatcca ccagggtatg catatggtag ctggccacga 1140tgccaatgat gctgtcattg ctaattcagt tgctcaggct cgcttttcag gactcctaat 1200tgtcaaaacc gttcttgatc atattctgca aaaaaccgac caaggagtaa gacttcaccc 1260tttggcccga acagccaaag tgcgtaatga ggttaatgca tttaaggccg ccctaagctc 1320acttgctaag catggggaat atgccccttt tgctcgcctt ctcaatctct cgggagttaa 1380caacctagaa catggtctct acccacagtt atcagcaatt gctcttggag ttgccacagc 1440acatggtagc acccttgcag gagttaatgt tggtgagcag tatcagcagc ttagagaggc 1500tgccactgaa gctgagaagc aactccaaca atatgctgag tccagagaac tcgacagcct 1560aggcctggac gatcaggaaa gaagaatact aatgaacttc catcagaaga aaaacgaaat 1620tagtttccag cagaccaatg caatggtaac ccttaggaaa gagcgactgg ctaaattaac 1680agaagctata acgctggcct caagacctaa cctcgggtct agacaagacg acggcaatga 1740aataccgttc cctgggccta taagcaacaa cccagaccaa gatcatctgg aggatgatcc 1800tagagactcc agagacacca tcattcctaa tggtgcaatt gaccccgagg atggtgattt 1860tgaaaattac aatggctatc atgatgatga agttgggacg gcaggtgact tggtcctgtt 1920cgatcttgac gatcatgagg atgacaataa agcttttgag ccacaggaca gctcgccaca 1980atcccaaagg gaaatagaga gagaaagatt aattcatcca cccccaggca acaacaagga 2040cgacaatcga gcctcagaca acaatcaaca atcagcagat tctgaggaac aaggaggtca 2100atacaactgg caccgaggcc cagaacgtac gaccgccaat cgaagactct caccagtgca 2160cgaagaggac acccttatgg atcaaggtga tgatgatccc tcaagcttac ctccgctgga 2220atctgatgat gacgatgcat caagtagcca acaagatccc gattatacag ctgttgcccc 2280tcctgctcct gtataccgca gtgcagaagc ccacgagcct ccccacaaat cctcgaacga 2340gccagctgaa acatcacaat tgaatgaaga ccctgatatc ggtcaatcaa agtctatgca 2400aaaattagaa gagacatatc accatctgct gagaactcaa ggtccatttg aagccatcaa 2460ttattatcac atgatgaagg atgagccggt aatatttagc actgatgatg ggaaggaata 2520cacctacccg gattcacttg aggaagccta tcctccatgg ctcaccgaga aagaacgact 2580ggacaaagag aatcgctaca tttacataaa taatcaacag ttcttctggc ctgtcatgag 2640tcccagagac aaatttcttg caatcttgca gcaccatcag taaccacagc acaaagcgcg 2700gtccacttcg taaagctaaa tacacttaag acttgaccga ttcatctaca aaaactaatc 2760cattataact tattagtgct acttttctat aagtgattct taatctaagg ccattaagag 2820tttaagtaat atacatatac acttacaccg gtctatccaa gatgtggctc aatgttcttg 2880atttgaacat agtcataagg ggataaataa tactttatat ttctgattgt ggattgaccc 2940attctgctta aaatgcttcg cccattgaaa atgtgatcta atagatagcc ctgactagac 3000aaattaagaa aaacatttga tgaagattaa aaccttcatc gccagtaaat gattatattg 3060tctgtaggca ggtgtttact ccaccttaaa tttggaaata tcctacctta ggaccattgt 3120caagaggtgc ataggcatta ccacccttga gaacatgtac aataataaat tgaaggtatg 3180ttcaggccca gaaacgactg gatggatttc tgagcaactt atgacaggta agattccagt 3240aactgatata ttcattgata ttgataacaa gccagatcaa atggaagtcc gactcaaacc 3300atcatcaagg agctcaacaa gaacttgtac aagtagcagt cagacggagg tcaactatgt 3360acctctcctt aaaaaggttg aggatacatt aactatgcta gtgaatgcca ccagtcgtca 3420gaatgctgca atcgaggccc ttgaaaaccg cctcagcaca cttgagagta gcttaaagcc 3480aatccaagac atgggtaaag tgatttcatc attgaatcgc agttgtgccg aaatggttgc 3540aaaatatgat cttctagtta tgacaactgg acgggctact tcaactgcag ctgcagtaga 3600tgcgtattgg aaagagcaca aacagccacc accagggcca gcgttgtatg aagagaatgc 3660gcttaaagga aaaatcgatg atccaaacag ctatgtacca gatgctgtgc aagaggctta 3720caagaacctt gacagtacat cgaccctgac cgaggaaaat tttgggaaac cttatatatc 3780tgctaaagac ctgaaggaga tcatgtatga tcatctacct ggttttggga ctgcctttca 3840ccaacttgtt caagtgattt gtaaaatagg aaaggataac aaccttttgg acacaatcca 3900tgctgagttc caggcaagtc tagcagatgg tgactctccc caatgtgcac tcatacagat 3960aaccaaaagg gtcccaatct ttcaggatgt gccgcccccg ataatccata ttagatcccg 4020tggtgacatc ccacgagcat gccaaaagag tctccgacca gcaccaccat cacccaaaat 4080tgatcgtggt tgggtttgtt tgtttaagat gcaagatggt aaaacgcttg gacttaagat 4140ctaagaatca agatttattt aacaaggcaa gccacaacct tagatggaac ctcagccaga 4200ctattgaact attgacgctg ttgatgataa tatataatta atggtcttat ttgaatatga 4260caacatcttg cttcttgttc tgccttgtag ctctttgaat tggaagatca ttccaaactt 4320acaaacatgc acaagatgtt atggtttagc aaagaattga taggagtact ggtatataat 4380gtaaatataa caagtgatga agattaagaa aaaccagtcg gtattttcca gacttggcat 4440ttcttatctt catcttctaa agtgagatat tttatcatca aaaaatgagg cgcggagtgt 4500taccaacggc tcctccagca tataatgata ttgcataccc tatgagcata ctcccaaccc 4560gaccaagtgt catagtcaat gagaccaaat cagatgtact ggcagtgcca ggggcagatg 4620ttccatcaaa ctccatgaga ccagtggctg atgataacat tgatcactca agccatactc 4680caagcggagt agcttctgcc tttatattgg aagctacagt gaatgtaatt tcgggaacaa 4740aagtcctgat gaagcaaata cctatttggc ttccactggg tgtagctgat cagaagatat 4800acagctttga ttcaacaaca gccgcaatta tgttggcttc ctacacagtg acacacttcg 4860ggaagatatc taacccgctg gtacgtgtca acaggctagg cccaggaata cccgatcatc 4920cgctacgact cctaaggttg ggcaatcagg cattccttca agagtttgtt cttccaccag 4980tccagcttcc ccagtatttc acatttgatc taacagctct aaagctcatc actcaaccat 5040tgccagctgc aacctggaca gacgaaactc cagcaggagc agtcaatgct cttcgtcctg 5100ggctctcact ccatcccaag cttcgtccaa ttctcctgcc ggggaagaca ggaaagaaag 5160gacatgcttc agacttaaca tcacctgaca agattcaaac aatcatgaat gcaataccgg 5220acctcaaaat tgtcccgatt gatccaacca agaacatagt tggaattgag gttccagaat 5280tactagttca aaggctgacc ggcaaaaaac cacaacccaa aaatggccaa ccaattattc 5340cagttcttct tccgaaatat gttggacttg atcctatatc gccaggggac ttaactatgg 5400ttatcaccca ggattgtgat tcatgccact ctccagccag ccatccgtat cacatggaca 5460agcagaatag ttaccaataa tttaaattcc attcgagcta ttattctgct agtaattccg 5520acgggatcaa tagactaaaa atctgattgt atagaattat aaaagaatca agcagaggca 5580acagactcac agcttacgcc tagataacta atattaagga gttttttaat ctaattttcc 5640agtcttgagt aataatcatt tcttttgtaa ttaattatgc atttgttaac ttatcggtgc 5700gagatttcct tgagaacccg gcggagcttc tactatctgc agtaaccaga agagaagttc 5760aacccagtca aaactaaacc aagcaatatt ctgaatgctc tatagtctat tctaatcaga 5820ggtataacaa tggctaagat ttcaatgact cgttaacaat cgctagtaat tttaatctcc 5880agattaagaa aaagatatac gatgaagatt aaggcgacaa cgagccgaaa cttcatctct 5940tttaaagatc taacattatc tgttccaaag tcatacaagg acacattcaa atcagggatt 6000gtaagctgct atttcttacc tccccaaatt acctatacaa catggggtca ggatatcaac 6060ttctccaatt gcctcgggaa cgttttcgta aaacttcgtt cttagtatgg gtaatcatcc 6120tcttccagcg agcaatctcc atgccgcttg gtatagtgac aaatagcact ctcaaagcaa 6180cagaaattga tcaattggtt tgtcgggaca aactgtcatc aaccagtcag ctcaagtctg 6240tggggctgaa tctggaagga aatggaattg caaccgatgt cccatcagca acaaaacgct 6300ggggatttcg ttcaggtgtg cctcccaagg tggtcagcta tgaagccgga gaatgggcag 6360aaaattgcta caatctggag atcaaaaagt cagacggaag tgaatgcctc cctctccctc 6420ccgacggtgt acgaggattc cctagatgtc gctatgtcca caaagttcaa ggaacaggtc 6480cttgtcctgg tgacttagct ttccataaaa atggggcttt tttcttgtat gatagattgg 6540cctcaactgt catctaccga gggacaactt ttgctgaagg tgtcgtagct tttttaattc 6600tgtcagagcc caagaagcat ttttggaagg ctacaccagc tcatgaaccg gtgaacacaa 6660cagatgattc cacaagctac tacatgaccc tgacactcag ctacgagatg tcaaattttg 6720ggggcaatga aagcaacacc ctttttaagg tagacaacca cacatatgtg caactagatc 6780gtccacacac tccgcagttc cttgttcagc tcaatgaaac acttcgaaga aataatcgcc 6840ttagcaacag tacagggaga ttgacttgga cattggatcc taaaattgaa ccagatgttg 6900gtgagtgggc cttctgggaa actaaaaaaa cttttcccaa caacttcatg gagaaaactt 6960gcatttccaa attccatcaa cccacaccaa caactcctca gatcagagcc cggcgggaac 7020tgtccaagga aaaattagct accacccacc cgccaacaac tccgagctgg ttccaacgga 7080ttcccctcca gtggtttcag tgctcactgc aggacggaca gaggaaatgt cgacccaagg 7140tctaaccaac ggagagacaa tcacaggttt caccgcgaac ccaatgacaa ccaccattgc 7200cccaagtcca accatgacaa gcgaggttga taacaatgta

ccaagtgaac agccgaacaa 7260cacagcatcc attgaagact cccccccatc ggcaagcaac gagacaattt accactccga 7320gatggatccg atccaaggct cgaacaactc cgcccagagc ccacagacca agaccacgcc 7380agcacccaca acatccccga tgacccagga cccgcaagag acggccaaca gcagcaaacc 7440aggaaccagc ccaggaagcg cagccggacc aagtcagccc ggactcacta taaatacagt 7500aagtaaggta gctgattcac tgagtcccac caggaaacaa aagcgatcgg ttcgacaaaa 7560caccgctaat aaatgtaacc cagatcttta ctattggaca gctgttgatg agggggcagc 7620agtaggattg gcatggattc catatttcgg acctgcagca gaaggcatct acattgaggg 7680tgtaatgcat aatcagaatg ggcttatttg cgggctacgt cagctagcca atgaaactac 7740ccaggctctt caattatttc tgcgggccac aacagaactg aggacttact cacttcttaa 7800cagaaaagct attgattttc ttcttcaacg atggggaggt acctgtcgaa tcctaggacc 7860atcttgttgc attgagccac atgattggac aaaaaatatt actgatgaaa ttaaccaaat 7920taaacatgac tttattgaca atcccctacc agaccacgga gatgatctta atctatggac 7980aggttggaga caatggatcc cggctggaat tgggattatt ggagttataa ttgctataat 8040agccctactt tgtatatgta agattttgtg ttgatttatt ctgagatctg agagagaaaa 8100atctcagggt tactctaagg agaaatatta tttttaaaat ttacttgaat gctgaccact 8160tatcttaaat gagcaattaa taatatgttt ttctgcttct ttgcttgatt tacaatatga 8220tatttctctt aataatgatt aatatattaa gaaaaactta tgacgaagat taaaggagag 8280gatcgttaac gggaaaacct cccatctcgt tcgtcgaagc cacgttggtg gtgcttgcag 8340ctgagaacaa ctccagagat tgtaggtaga aaggaccaac atttataggt aggggtcaga 8400aagcaacaat aaccataaaa ggagagcctg acattgctat ttaatatcct agaacctgat 8460ttctaggttc tagctttaaa atccggatga tggagcattc aagagaacgg ggtagatcta 8520gcaacatgcg acataatagc cgggaaccat acgaaaatcc atcaaggtct cgctcattat 8580ctcgggaccc taatcaggtt gatcgtagac agcctcgaag tgcatcccaa attcgtgttc 8640cgaatctgtt ccatcggaaa aagactgatg cactcatagt tcctccggct cctaaagata 8700tatgcccaac actcaaaaaa ggattcctct gcgatagcaa attttgcaaa aaagatcacc 8760aattggatag cttaaatgat catgaattac tactgctaat tgcaagaaga acatgtggaa 8820ttatcgagag caattcgcag attacatccc caaaagatat gcggttagcg aatccaacag 8880ctgaagactt ctcacaaggt aatagtccta aattaacact tgcagtcctt cttcaaattg 8940ctgaacattg ggcaaccaga gacctaaggc aaattgagga ctctaaactt agagctcttt 9000taaccctttg tgccgtatta acaaggaaat tttctaaatc ccaactgggt cttctatgtg 9060agacccacct acggcatgag ggcctcggac aggaccaagc tgattctgta ttagaggtct 9120accaaagact ccacagtgat aaaggaggga attttgaggc tgccctgtgg caacaatggg 9180accgacagtc attaataatg ttcatctctg cttttctcaa cattgctctc cagatacctt 9240gtgaaagttc tagtgtcgta gtctcaggtc ttgccacatt gtacccagca caagacaatt 9300ctacaccatc cgaggcaact aatgatacca cctggtcaag tacagttgaa tagaaaacca 9360ctggagctat ttttccacga ttgctctcag tcaataaatt aatatagata taatacgact 9420tcggtgtgca attgtcaaga gttccattta gtaataatga ttcttaaaac aatctactat 9480cgcaattatc gatggatcta ccctatttga cggtacatga cttgaatgta ataaggtaag 9540ttggtatctg aggtattttg tctagagtat actcaaaatc gtatgtctag caaattatca 9600atagcaaagt taaattctcc taacctcata ttttgatcaa gtaatcatga ttttatgata 9660attcttttca gattatcggt ttaatcttta ttaagaaaaa atcatgattg tagacaattt 9720actggtagtc cttgggtatc caagtttatg aatagagcta gagagaattt gctacttccg 9780aggtataact ttattatttg ctacttcgaa tgcctaaaac cagtaatgca ggatgaagat 9840taattgcgga ggaatcagga attcaacttt agttccttaa ggcctcgtcc gaatcttcat 9900cagttcgtaa gttcttttat agaagtcatt agcttctaag gtgattatat tttagtatta 9960aattttgcta attgcttgct ataaagttga aatgtctaat gcttaaatga acactttttt 10020gaagctgaca tacgaataca tcatatcata tgaaaacatc gcaattagag cgtccttgaa 10080gtctggcatt gacagtcacc aggctgttct cagtagtctg tccttggaag ctcttgggga 10140gacaaaaaga ggtcccagag agtcccaaca ggttggcata aggtcattaa caccagcata 10200gtcggctcga ccaagactgt aagcgagtcg atttcaacta aaaagattat ttcttgttgt 10260ttaaacaaat tccttttgtg tgagacatcc tcaaggcaca agatggctaa agccacaggc 10320cgatacaatc tcgtgccccc aaagaaagat atggaaaagg gagtgatttt tagtgatctt 10380tgtaatttct tgattactca aaccctgcaa ggttggaagg tttattgggc aggaattgag 10440tttgatgtaa gtcaaaaagg catggctctt ctgacaagac tcaaaacaaa tgactttgct 10500cctgcctggg cgatgacaag aaatcttttc ccacatctgt tccagaaccc aaattcggtt 10560attcaatctc ccatctgggc tttgagggta attttggcag ccggattgca ggatcagttg 10620ttagaccatt cattggttga gccattgaca ggggctctcg gtctaatttc tgattggctc 10680ctaactacaa cgtcaacaca tttcaatctt cgtactagaa gcgtaaagga ccagcttagt 10740cttcgtatgt tatctttgat caggtcaaac atcttgcagt tcatcaacaa gcttgacgcc 10800ctgcatgttg tcaattacaa tggtttactc agtagtattg agatcgggac ttctacacac 10860acaatcatta taactcgtac aaatatgggt tttctcgtgg aagttcaaga gcctgacaaa 10920tcagctatga attctaagcg cccaggacca gtcaagttct cattacttca tgagtctgcc 10980ttcaaacctt tcactcgtgt tccacaatct gggatgcaat cattaataat ggagttcaac 11040agtttgttgg caatttaaca aggtaatctt aaaataagta catgaatgag aattagttgt 11100gggtcttatc tagcattgtt gagttaacta tctaatctat tttcgctaat tgcattgagc 11160actgctaata ggtttgtatc acgttaaaga tttagagtgt atgaattgtg cagatttaaa 11220cttgggtttt gccttatgct tcataggtgg tctttttgaa atggagatta tcagcatttc 11280ttaaatggga ggagttagca atcagaaatt ggagataaat ggacatcggg atagaacaat 11340gcctaactat tgggcggctt ccatttttac atgtgtatat aaccaatctt ttcctatctt 11400tgcttatatt ggtgtaactt tattttaata acatgtcaat gctatactgt taagagaagg 11460tctgaggaag attaagaaaa aggcctcgtg ttcacttggt tgccgtcaag tatcctgtgg 11520tttttttcta cctaacttcc tcatgccata tggctaccca gcatacccag tacccggatg 11580cacgtttatc ctcacctata gtcctggatc aatgtgattt ggtaactcga gcatgtgggt 11640tatattcatc ttattctcta aatcctcaac taaggcaatg taaattacca aaacatatat 11700atcgacttaa gttcgacaca atagtatcca aattcctaag tgatacacct gtagcaacac 11760tgccgataga ctatttagta ccaattctcc tgcgttccct aacggggcac ggtgataggc 11820cattgacccc gacttgcaat caattccttg atgaaattat taattacact cttcatgatg 11880cagcctttct tgattactat ctcaaggcaa caggtgcaca ggaccatttg acaaacattg 11940caactagaga gaagcttaaa aacgaaattc taaacaatga ttatgtccat caattgttct 12000tctggcatga cctttctatt ttggctcgac gtgggcgtct gaatcgcggg aacaaccgtt 12060caacctggtt tgttcatgat gaattcattg atattttagg atatggcgat tatatttttt 12120ggaaaatacc tttatcatta ttaccagtta ctatagacgg ggtcccacac gcagcaactg 12180actggtatca accgactctt tttaaagaat ccatcctagg gcatagccaa atcctatctg 12240tgtcaacagc tgaaatacta attatgtgta aagatattat cacctgtagg tttaatacat 12300cactgattgc atccattgca aaattagagg atgtagatgt gtctgattat cctgacccga 12360gtgatattct taagatatac aatgctggag actatgtaat atctattctt ggctcagagg 12420gttataagat aataaagtac cttgaaccac tttgtttggc caaaatccaa ctttgctcta 12480aattcacaga aagaaaaggt cgtttcctca cacagatgca tttatcagta ataaatgatc 12540ttcgggagtt gatttctaac cgcaggttaa aggactatca gcaagagaag attagagatt 12600ttcacaaaat attattacaa ttgcaattat ctcctcaaca gttttgtgaa ttattctctg 12660ttcaaaaaca ttgggggcat ccaattttac atagtgagaa agctatacaa aaagtaaaac 12720ggcatgcaac catccttaag gctctcagac ctaatgtcat ctttgagaca tattgtgtat 12780tcaagtacaa tattgccaag cactatttcg acagccaagg aacttggtac agtgtaatct 12840cagacaggaa tttaactcca ggactcaact ccttcataaa acgtaatcac tttccttcac 12900tacccatgat taaggatctt ctatgggaat tctatcatct taatcaccct ccgttattct 12960ctacaaaggt gattagtgac ttaagtattt tcatcaaaga tagggccaca gctgttgaac 13020agacatgttg ggatgcagtc tttgaaccca atgtgctagg ttacaatcct ccaaacaaat 13080tctccactaa aagggtgccg gaacaatttc tagaacaaga ggatttttca atcgaaagtg 13140tcctgaatta tgcacaggaa ttacattatt tattaccaca gaataggaat ttttcctttt 13200ctctcaaaga aaaggaatta aatattggac gaacatttgg gaagctacca tatctcacac 13260ggaatgtcca aactttatgt gaggctctgt tagcagatgg actggccaag gccttcccca 13320gtaacatgat ggtagtaact gaacgtgaac aaaaagagag ccttcttcat caggcatcat 13380ggcaccacac cagtgatgat tttggagaga atgctaccgt tcgagggagt agttttgtaa 13440ctgatttaga gaagtacaat cttgcatttc gctatgagtt cactgcacca tttattgagt 13500actgcaacca ttgctatggt gtgcgtaatg tctttaattg gatgcattat ttaatcccgc 13560agtgttacat gcatgtaagt gattattata atccgcctca caatgttaat cttagcaatc 13620gagaatatcc tcctgaaggc ccgagttcgt accgagggca cttaggaggc atagagggat 13680tacaacaaaa actgtggacg agtatatcct gtgcacaaat ctccttagtg gaaattaaaa 13740ctggttttaa gttacgatca gcggtcatgg gagacaatca gtgtataacc gtattgtctg 13800tttttccact taaaacagac cctgaagagc aggagcaaag cgccgaagac aatgctgcaa 13860gagtagcagc aagtcttgca aaagtaacca gtgcatgtgg gatctttctt aaaccagatg 13920agacatttgt acactcaggt ttcatttatt tcggaaaaaa acaatatctc aatggtgtac 13980aattaccgca atcactcaaa acagcagcaa gaatggcacc actctctgat gctatattcg 14040atgatctaca aggaacactt gccagtattg gaactgcctt cgaacgtgct atatcggaaa 14100cgcgacatat cctcccatgt cgtattgtag cagctttcca tacgtatttc gccgttcgga 14160ttttacaata tcaccatctt ggatttaata aaggcatcga tttaggacag ttgtcactta 14220gtaaaccatt agactatggg actattactc taacattggc ggttccacaa gtccttgggg 14280gattgtcttt tctaaatcca gaaaagtgtt tttatcgaaa cttcggagat cctgtgactt 14340ctggactttt ccagctacgg gtgtacctag aaatggttaa catgaaagac ctattttgtc 14400cattaatatc gaaaaatcca ggaaattgta gtgccattga ttttgtctta aatccatccg 14460gattaaatgt tccaggatca caagacttga catccttttt gcgacaaatc gttaggcgta 14520gtattaccct aactgctaga aataagttaa ttaacactct cttccatgcc tctgctgatt 14580tggaagatga gatggtttgt aagtggctcc tttcatcaaa ccctgtcatg agtcgctttg 14640cagcggatat tttttccagg acaccgagtg gtaaacgtct ccaaatatta ggttatcttg 14700aagggaccag gactctattg gcctccaaaa tcataaacaa caacagtgag acacctgtac 14760ttgataagct gaggaagatc accctacaaa gatggaatct gtggttcagt tatttggacc 14820attgtgacca attactagca gatgctctac agaaaattag ttgcacggtg gatttggccc 14880agattttgcg tgagtataca tggtcacaca tcttagaggg tagatcattg atcggagcga 14940cattaccatg tatggtggag caattcaaag ttaagtggct aggacaatat gaaccttgtc 15000cagaatgtct caacaaaaaa ggctcaaatg cttatgtctc agttgcagtc aaagatcaag 15060tggtcagtgc ttggcctaat acttctcgaa taagttggac aatagggagt ggtgtcccct 15120atatagggtc aagaaccgag gataaaatcg gacagcctgc tatcaagccg cgatgccctt 15180catctgccct caaggaggct atagaattag catcaaggct cacttgggtt acacaaggag 15240gttctaatag tgaacaatta atccggcctt tcttggaagc gagagtcaac cttagtgtca 15300gtgaagtcct gcaaatgaca ccatcacatt attcaggaaa tattgtccat cgatataacg 15360accaatacag cccgcactca tttatggcga atcgcatgag caatactgcg acccgtctca 15420tagtgtcaac taatacactt ggagaatttt caggtggagg gcaggccgcc agggatagca 15480atataatttt ccagaatgtt ataaatttag cagttgccct ttatgatatt agattccgga 15540atacaaacac ctctgatata aggcataata gggctcatct tcacctgaca gagtgctgta 15600ctaaagaggt cccggcccag tatttgacat atacaagtgc acttaatctg gatttaagcc 15660gttatcgtga taatgaacta atatatgact caaatccact gaagggagga ttgaactgca 15720atttaacaat agatagtcct ttagtgaagg gtcctaggct taacatgatt gaagatgatc 15780ttctccgctt tccacacctt tctggatggg agttagcgaa aacggtggta caatccatca 15840tctcagacaa tagcaactca tcaacagatc caatcagtag cggagaaaca cgctctttca 15900caactcattt tctcacttac cctcagattg gccttcttta cagtttcgga gcagtattat 15960gcttttatct aggcaatact atcctatgga ctaaaaaact tgattacgaa cagtttctat 16020attatttgca taaccagctg cacaacttac ctcatcgagc actccgtgtt tttaaaccaa 16080catttaagca tgccagtgtg atgtcccgat taatggaaat tgattctaac ttctcaattt 16140atattggcgg gacatctgga gatcgagggc tgtctgatgc tgctcgactg tttcttcgga 16200cagcaatcgc gagtttttta caatttctta aaagctggat catcgatcgc caaaagacaa 16260ttcctttatg gatagtatat ccgcttgaag gtcaacagcc ggaatccatc aatgaatttc 16320tacataaaat tttgggtctg ctcaaacaag gccccaaaag tattccaaag gaggtcagca 16380tccaaaatga tggacatttg gatttggcag aaaataatta tgtttacaat agtaagagca 16440ctgctagtaa tttcttccat gcatccttag cttactggag aagtaggaaa tctcggaaaa 16500ctcaagacca taatgatttc tcaagagggg atggaacact tacagaaccc gtgcgtaagt 16560tttcaagcaa tcatcagtca gatgaaaagt actacaatgt gacatgtgga aagtcaccga 16620agccgcaaga acgcaaagac ttctcgcaat acagactcag caataacggg caaacaatga 16680gtaatcatcg taagaaaggg aagttccaca agtggaatcc ctgcaaaatg ttaatggaga 16740gtcaaagggg aactgttcta acagagggtg actactttca aaacaatact ccaccaacag 16800atgatgtatc aagtcctcac cgactcattc taccattttt taaattggga aatcacaacc 16860atgcacatga tcaagatgcc caagaattga tgaatcaaaa tattaaacag tacctacatc 16920agctaaggtc tatgttggac accactatat attgtagatt cacagggatt gtctcatcca 16980tgcattacaa attggacgaa gttcttctag aatacaatag tttcgattca gctatcacat 17040tagctgaagg tgaggggtca ggggctctat tacttttgca aaaatatagt acaaggttat 17100tatttttgaa cacattggca acagaacaca gtatagaatc agaagttgta tcaggttttt 17160ctactccgag aatgttgtta ccaataatgc aaaaggttca tgaaggacaa gtcactgtta 17220tcttaaataa ttcagcaagt cagataactg acataactag ctcaatgtgg ttaagtaatc 17280aaaaatataa tctaccttgt caagttgaaa tcattatgat ggatgctgaa acaacagaga 17340acttaaacag gtcccaactc taccgagcag tatataactt aatacttgat cacattgatc 17400cgcagtatct caaggtggtg gtactcaaag tatttctgag tgatatagaa ggaatattat 17460ggattaatga ttacttggct ccattattcg gggctggtta cttgattaaa ccgattacat 17520caagtgcccg gtcaagtgaa tggtaccttt gcttatcaaa tttgatatct actaacagga 17580gatcggccca tcagactcac aaggcatgtc ttggtgttat cagagatgct ttgcaagcac 17640aagtccagcg aggcgtgtac tggttgagtc acatcgcaca gtatgctaca aagaatctcc 17700attgtgaata cataggcctt ggtttcccat ctctagaaaa ggtcctatat cacaggtata 17760atctagttga tactggactc ggtccattgt cgtcagttat tagacattta actaacctcc 17820aggcagagat acgagactta gtattagatt ataacctgat gagggagagt cgcactcaaa 17880cgtaccattt tattaagact gcaaaaggca gaatcacaaa gttagtcaat gactttctga 17940agttttcttt aattgtccag gcactcaaaa ataattcttc ttggtatact gagcttaaaa 18000aattacctga ggttattaat gtgtgtaatc gattttatca tactcacaat tgcgaatgtc 18060aggaaaaatt ctttgtccag acgctttatt tacaacgcct acgcgatgca gaaatcaagc 18120taattgaacg ccttaccggg ttaatgcgat tttatccaga agggttaata tattccaatc 18180acacataggt actaaatcat catagtatga ggaataagat aatgataatt cctgacgaca 18240gttttagttc cgattctaag tatatcggaa gagagtatgc caatcttaat tgttagaggt 18300aacaagctat tagttattac ttattgataa gaatacactt tatcatagcg taacacatca 18360taactttata acgattttgc atttctaatc ctagtattta ttagaatgta ctaccagaga 18420aatgacccca gttcctatct ttaaataatg attgtgtgta ttaaattatt agtttattag 18480gtttatgagt tggttacaca gtgagtatta gtaattgagg attatgtaga taggtaatct 18540aacactgaat cacccatctg atgtcaccat atccaaatgt tgtgctagtc gcatttaaac 18600atgctatctt cagttaagta acatagactg aaaatgctaa gaagagattg gagtaaaagt 18660ataaaataaa tttaattaaa cttcaaagtg attaaatgat aatgatcttg ggaactcgat 18720atgacctcaa gtcaaaaata atgtcaatat aattgtttag taatatgagt gataatgtaa 18780attttgataa ctaactagct ttagtagtta agatcaaatg caaacattat aagaatgtta 18840agcgcacaca aaaacattat aaaaaaccaa ttttttcctt tttgtgtgtc c 1889110287PRTReston ebolavirus 10Met Glu His Ser Arg Glu Arg Gly Arg Ser Ser Asn Met Arg His Asn1 5 10 15 Ser Arg Glu Pro Tyr Glu Asn Pro Ser Arg Ser Arg Ser Leu Ser Arg 20 25 30 Asp Pro Asn Gln Val Asp Arg Arg Gln Pro Arg Ser Ala Ser Gln Ile 35 40 45 Arg Val Pro Asn Leu Phe His Arg Lys Lys Thr Asp Ala Leu Ile Val 50 55 60 Pro Pro Ala Pro Lys Asp Ile Cys Pro Thr Leu Lys Lys Gly Phe Leu65 70 75 80 Cys Asp Ser Lys Phe Cys Lys Lys Asp His Gln Leu Asp Ser Leu Asn 85 90 95 Asp His Glu Leu Leu Leu Leu Ile Ala Arg Arg Thr Cys Gly Ile Ile 100 105 110 Glu Ser Asn Ser Gln Ile Thr Ser Pro Lys Asp Met Arg Leu Ala Asn 115 120 125 Pro Thr Ala Glu Asp Phe Ser Gln Gly Asn Ser Pro Lys Leu Thr Leu 130 135 140 Ala Val Leu Leu Gln Ile Ala Glu His Trp Ala Thr Arg Asp Leu Arg145 150 155 160 Gln Ile Glu Asp Ser Lys Leu Arg Ala Leu Leu Thr Leu Cys Ala Val 165 170 175 Leu Thr Arg Lys Phe Ser Lys Ser Gln Leu Gly Leu Leu Cys Glu Thr 180 185 190 His Leu Arg His Glu Gly Leu Gly Gln Asp Gln Ala Asp Ser Val Leu 195 200 205 Glu Val Tyr Gln Arg Leu His Ser Asp Lys Gly Gly Asn Phe Glu Ala 210 215 220 Ala Leu Trp Gln Gln Trp Asp Arg Gln Ser Leu Ile Met Phe Ile Ser225 230 235 240 Ala Phe Leu Asn Ile Ala Leu Gln Ile Pro Cys Glu Ser Ser Ser Val 245 250 255 Val Val Ser Gly Leu Ala Thr Leu Tyr Pro Ala Gln Asp Asn Ser Thr 260 265 270 Pro Ser Glu Ala Thr Asn Asp Thr Thr Trp Ser Ser Thr Val Glu 275 280 285 1118960DNAZaire ebolavirus 11cggacacaca aaaagaaaga agaattttta ggatcttttg tgtgcgaata actatgagga 60agattaataa ttttcctctc attgaaattt atatcggaat ttaaattgaa attgttactg 120taatcacacc tggtttgttt cagagccaca tcacaaagat agagaacaac ctaggtctcc 180gaagggagca agggcatcag tgtgctcagt tgaaaatccc ttgtcaacac ctaggtctta 240tcacatcaca agttccacct cagactctgc agggtgatcc aacaacctta atagaaacat 300tattgttaaa ggacagcatt agttcacagt caaacaagca agattgagaa ttaaccttgg 360ttttgaactt gaacacttag gggattgaag attcaacaac cctaaagctt ggggtaaaac 420attggaaata gttaaaagac aaattgctcg gaatcacaaa attccgagta tggattctcg 480tcctcagaaa atctggatgg cgccgagtct cactgaatct gacatggatt accacaagat 540cttgacagca ggtctgtccg ttcaacaggg gattgttcgg caaagagtca tcccagtgta 600tcaagtaaac aatcttgaag aaatttgcca acttatcata caggcctttg aagcaggtgt 660tgattttcaa gagagtgcgg acggtttcct tctcatgctt tgtcttcatc atgcgtacca 720gggagattac aaacttttct tggaaagtgg cgcagtcaag tatttggaag ggcacgggtt 780ccgttttgaa gtcaagaagc gtgatggagt gaagcgcctt gaggaattgc tgccagcagt 840atctagtgga aaaaacatta agagaacact tgctgccatg ccggaagagg agacaactga 900agctaatgcc ggtcagtttc tctcctttgc aagtctattc cttccgaaat tggtagtagg 960agaaaaggct tgccttgaga aggttcaaag gcaaattcaa gtacatgcag agcaaggact 1020gatacaatat ccaacagctt ggcaatcagt aggacacatg atggtgattt tccgtttgat 1080gcgaacaaat tttctgatca aatttctcct aatacaccaa gggatgcaca tggttgccgg 1140gcatgatgcc aacgatgctg tgatttcaaa ttcagtggct caagctcgtt tttcaggctt 1200attgattgtc aaaacagtac ttgatcatat cctacaaaag acagaacgag gagttcgtct 1260ccatcctctt gcaaggaccg ccaaggtaaa aaatgaggtg aactccttta aggctgcact 1320cagctccctg gccaagcatg gagagtatgc tcctttcgcc cgacttttga acctttctgg 1380agtaaataat cttgagcatg gtcttttccc tcaactatcg gcaattgcac tcggagtcgc 1440cacagcacac gggagtaccc tcgcaggagt aaatgttgga gaacagtatc

aacaactcag 1500agaggctgcc actgaggctg agaagcaact ccaacaatat gcagagtctc gcgaacttga 1560ccatcttgga cttgatgatc aggaaaagaa aattcttatg aacttccatc agaaaaagaa 1620cgaaatcagc ttccagcaaa caaacgctat ggtaactcta agaaaagagc gcctggccaa 1680gctgacagaa gctatcactg ctgcgtcact gcccaaaaca agtggacatt acgatgatga 1740tgacgacatt ccctttccag gacccatcaa tgatgacgac aatcctggcc atcaagatga 1800tgatccgact gactcacagg atacgaccat tcccgatgtg gtggttgatc ccgatgatgg 1860aagctacggc gaataccaga gttactcgga aaacggcatg aatgcaccag atgacttggt 1920cctattcgat ctagacgagg acgacgagga cactaagcca gtgcctaata gatcgaccaa 1980gggtggacaa cagaagaaca gtcaaaaggg ccagcatata gagggcagac agacacaatc 2040caggccaatt caaaatgtcc caggccctca cagaacaatc caccacgcca gtgcgccact 2100cacggacaat gacagaagaa atgaaccctc cggctcaacc agccctcgca tgctgacacc 2160aattaacgaa gaggcagacc cactggacga tgccgacgac gagacgtcta gccttccgcc 2220cttggagtca gatgatgaag agcaggacag ggacggaact tccaaccgca cacccactgt 2280cgccccaccg gctcccgtat acagagatca ctctgaaaag aaagaactcc cgcaagacga 2340gcaacaagat caggaccaca ctcaagaggc caggaaccag gacagtgaca acacccagtc 2400agaacactct tttgaggaga tgtaccgcca cattctaaga tcacaggggc catttgatgc 2460tgttttgtat tatcatatga tgaaggatga gcctgtagtt ttcagtacca gtgatggcaa 2520agagtacacg tatccagact cccttgaaga ggaatatcca ccatggctca ctgaaaaaga 2580ggctatgaat gaagagaata gatttgttac attggatggt caacaatttt attggccggt 2640gatgaatcac aagaataaat tcatggcaat cctgcaacat catcagtgaa tgagcatgga 2700acaatgggat gattcaaccg acaaatagct aacattaagt agtcaaggaa cgaaaacagg 2760aagaattttt gatgtctaag gtgtgaatta ttatcacaat aaaagtgatt cttatttttg 2820aatttaaagc tagcttatta ttactagccg tttttcaaag ttcaatttga gtcttaatgc 2880aaataggcgt taagccacag ttatagccat aattgtaact caatattcta actagcgatt 2940tatctaaatt aaattacatt atgcttttat aacttaccta ctagcctgcc caacatttac 3000acgatcgttt tataattaag aaaaaactaa tgatgaagat taaaaccttc atcatcctta 3060cgtcaattga attctctagc actcgaagct tattgtcttc aatgtaaaag aaaagctggt 3120ctaacaagat gacaactaga acaaagggca ggggccatac tgtggccacg actcaaaacg 3180acagaatgcc aggccctgag ctttcgggct ggatctctga gcagctaatg accggaagaa 3240ttcctgtaag cgacatcttc tgtgatattg agaacaatcc aggattatgc tacgcatccc 3300aaatgcaaca aacgaagcca aacccgaaga cgcgcaacag tcaaacccaa acggacccaa 3360tttgcaatca tagttttgag gaggtagtac aaacattggc ttcattggct actgttgtgc 3420aacaacaaac catcgcatca gaatcattag aacaacgcat tacgagtctt gagaatggtc 3480taaagccagt ttatgatatg gcaaaaacaa tctcctcatt gaacagggtt tgtgctgaga 3540tggttgcaaa atatgatctt ctggtgatga caaccggtcg ggcaacagca accgctgcgg 3600caactgaggc ttattgggcc gaacatggtc aaccaccacc tggaccatca ctttatgaag 3660aaagtgcgat tcggggtaag attgaatcta gagatgagac cgtccctcaa agtgttaggg 3720aggcattcaa caatctaaac agtaccactt cactaactga ggaaaatttt gggaaacctg 3780acatttcggc aaaggatttg agaaacatta tgtatgatca cttgcctggt tttggaactg 3840ctttccacca attagtacaa gtgatttgta aattgggaaa agatagcaac tcattggaca 3900tcattcatgc tgagttccag gccagcctgg ctgaaggaga ctctcctcaa tgtgccctaa 3960ttcaaattac aaaaagagtt ccaatcttcc aagatgctgc tccacctgtc atccacatcc 4020gctctcgagg tgacattccc cgagcttgcc agaaaagctt gcgtccagtc ccaccatcgc 4080ccaagattga tcgaggttgg gtatgtgttt ttcagcttca agatggtaaa acacttggac 4140tcaaaatttg agccaatctc ccttccctcc gaaagaggcg aataatagca gaggcttcaa 4200ctgctgaact atagggtacg ttacattaat gatacacttg tgagtatcag ccctggataa 4260tataagtcaa ttaaacgacc aagataaaat tgttcatatc tcgctagcag cttaaaatat 4320aaatgtaata ggagctatat ctctgacagt attataatca attgttatta agtaacccaa 4380accaaaagtg atgaagatta agaaaaacct acctcggctg agagagtgtt ttttcattaa 4440ccttcatctt gtaaacgttg agcaaaattg ttaaaaatat gaggcgggtt atattgccta 4500ctgctcctcc tgaatatatg gaggccatat accctgtcag gtcaaattca acaattgcta 4560gaggtggcaa cagcaataca ggcttcctga caccggagtc agtcaatggg gacactccat 4620cgaatccact caggccaatt gccgatgaca ccatcgacca tgccagccac acaccaggca 4680gtgtgtcatc agcattcatc cttgaagcta tggtgaatgt catatcgggc cccaaagtgc 4740taatgaagca aattccaatt tggcttcctc taggtgtcgc tgatcaaaag acctacagct 4800ttgactcaac tacggccgcc atcatgcttg cttcatacac tatcacccat ttcggcaagg 4860caaccaatcc acttgtcaga gtcaatcggc tgggtcctgg aatcccggat catcccctca 4920ggctcctgcg aattggaaac caggctttcc tccaggagtt cgttcttccg ccagtccaac 4980taccccagta tttcaccttt gatttgacag cactcaaact gatcacccaa ccactgcctg 5040ctgcaacatg gaccgatgac actccaacag gatcaaatgg agcgttgcgt ccaggaattt 5100catttcatcc aaaacttcgc cccattcttt tacccaacaa aagtgggaag aaggggaaca 5160gtgccgatct aacatctccg gagaaaatcc aagcaataat gacttcactc caggacttca 5220agatcgttcc aattgatcca accaaaaata tcatgggaat cgaagtgcca gaaactctgg 5280tccacaagct gaccggtaag aaggtgactt ctaaaaatgg acaaccaatc atccctgttc 5340ttttgccaaa gtacattggg ttggacccgg tggctccagg agacctcacc atggtaatca 5400cacaggattg tgacacgtgt cattctcctg caagtcttcc agctgtgatt gagaagtaat 5460tgcaataatt gactcagatc cagttttata gaatcttctc agggatagtg ataacatcta 5520tttagtaatc cgtccattag aggagacact tttaattgat caatatacta aaggtgcttt 5580acaccattgt cttttttctc tcctaaatgt agaacttaac aaaagactca taatatactt 5640gtttttaaag gattgattga tgaaagatca taactaataa cattacaaat aatcctacta 5700taatcaatac ggtgattcaa atgttaatct ttctcattgc acatactttt tgcccttatc 5760ctcaaattgc ctgcatgctt acatctgagg atagccagtg tgacttggat tggaaatgtg 5820gagaaaaaat cgggacccat ttctaggttg ttcacaatcc aagtacagac attgcccttc 5880taattaagaa aaaatcggcg atgaagatta agccgacagt gagcgtaatc ttcatctctc 5940ttagattatt tgttttccag agtaggggtc gtcaggtcct tttcaatcgt gtaaccaaaa 6000taaactccac tagaaggata ttgtggggca acaacacaat gggcgttaca ggaatattgc 6060agttacctcg tgatcgattc aagaggacat cattctttct ttgggtaatt atccttttcc 6120aaagaacatt ttccatccca cttggagtca tccacaatag cacattacag gttagtgatg 6180tcgacaaact agtttgtcgt gacaaactgt catccacaaa tcaattgaga ccagttggac 6240tgaatctcga agggaatgga gtggcaactg acgtgccatc tgcaactaaa agatggggct 6300tcaggtccgg tgtcccacca aaggtggtca attatgaagc tggtgaatgg gctgaaaact 6360gctacaatct tgaaatcaaa aaacctgacg ggagtgagtg tctaccagca gcgccagacg 6420ggattcgggg cttcccccgg tgccggtatg tgcacaaagt atcaggaacg ggaccgtgtg 6480ccggagactt tgccttccat aaagagggtg ctttcttcct gtatgatcga cttgcttcca 6540cagttatcta ccgaggaacg actttcgctg aaggtgtcgt tgcatttctg atactgcccc 6600aagctaagaa ggacttcttc agctcacacc ccttgagaga gccggtcaat gcaacggagg 6660acccgtctag tggctactat tctaccacaa ttagatatca ggctaccggt tttggaacca 6720atgagacaga gtacttgttc gaggttgaca atttgaccta cgtccaactt gaaccaagat 6780tcacaccaca gtttctgctc cagctgaatg agacaatata tacaagtggg aaaaggagca 6840ataccacggg aaaactaatt tggaaggtca accccgaaat tgatacaaca atcggggagt 6900gggccttctg ggaaactaaa aaaacctcac tagaaaaatt cgcagtgaag agttgtcttt 6960cacagttgta tcaaacggag ccaaaaacat cagtggtcag agtccggcgc gaacttcttc 7020cgacccaggg accaacacaa caactgaaga ccacaaaatc atggcttcag aaaattcctc 7080tgcaatggtt caagtgcaca gtcaaggaag ggaagctgca gtgtcgcatc taacaaccct 7140tgccacaatc tccacgagtc cccaatccct cacaaccaaa ccaggtccgg acaacagcac 7200ccataataca cccgtgtata aacttgacat ctctgaggca actcaagttg aacaacatca 7260ccgcagaaca gacaacgaca gcacagcctc cgacactccc tctgccacga ccgcagccgg 7320acccccaaaa gcagagaaca ccaacacgag caagagcact gacttcctgg accccgccac 7380cacaacaagt ccccaaaacc acagcgagac cgctggcaac aacaacactc atcaccaaga 7440taccggagaa gagagtgcca gcagcgggaa gctaggctta attaccaata ctattgctgg 7500agtcgcagga ctgatcacag gcgggagaag aactcgaaga gaagcaattg tcaatgctca 7560acccaaatgc aaccctaatt tacattactg gactactcag gatgaaggtg ctgcaatcgg 7620actggcctgg ataccatatt tcgggccagc agccgaggga atttacacag aggggctaat 7680gcacaatcaa gatggtttaa tctgtgggtt gagacagctg gccaacgaga cgactcaagc 7740tcttcaactg ttcctgagag ccacaactga gctacgcacc ttttcaatcc tcaaccgtaa 7800ggcaattgat ttcttgctgc agcgatgggg cggcacatgc cacattctgg gaccggactg 7860ctgtatcgaa ccacatgatt ggaccaagaa cataacagac aaaattgatc agattattca 7920tgattttgtt gataaaaccc ttccggacca gggggacaat gacaattggt ggacaggatg 7980gagacaatgg ataccggcag gtattggagt tacaggcgtt ataattgcag ttatcgcttt 8040attctgtata tgcaaatttg tcttttagtt tttcttcaga ttgcttcatg gaaaagctca 8100gcctcaaatc aatgaaacca ggatttaatt atatggatta cttgaatcta agattacttg 8160acaaatgata atataataca ctggagcttt aaacatagcc aatgtgattc taactccttt 8220aaactcacag ttaatcataa acaaggtttg acatcaatct agttatctct ttgagaatga 8280taaacttgat gaagattaag aaaaaggtaa tctttcgatt atctttaatc ttcatccttg 8340attctacaat catgacagtt gtctttagtg acaagggaaa gaagcctttt tattaagttg 8400taataatcag atctgcgaac cggtagagtt tagttgcaac ctaacacaca taaagcattg 8460gtcaaaaagt caatagaaat ttaaacagtg agtggagaca acttttaaat ggaagcttca 8520tatgagagag gacgcccacg agctgccaga cagcattcaa gggatggaca cgaccaccat 8580gttcgagcac gatcatcatc cagagagaat tatcgaggtg agtaccgtca atcaaggagc 8640gcctcacaag tgcgcgttcc tactgtattt cataagaaga gagttgaacc attaacagtt 8700cctccagcac ctaaagacat atgtccgacc ttgaaaaaag gatttttgtg tgacagtagt 8760ttttgcaaaa aagatcacca gttggagagt ttaactgata gggaattact cctactaatc 8820gcccgtaaga cttgtggatc agtagaacaa caattaaata taactgcacc caaggactcg 8880cgcttagcaa atccaacggc tgatgatttc cagcaagagg aaggtccaaa aattaccttg 8940ttgacactga tcaagacggc agaacactgg gcgagacaag acatcagaac catagaggat 9000tcaaaattaa gagcattgtt gactctatgt gctgtgatga cgaggaaatt ctcaaaatcc 9060cagctgagtc ttttatgtga gacacaccta aggcgcgagg ggcttgggca agatcaggca 9120gaacccgttc tcgaagtata tcaacgatta cacagtgata aaggaggcag ttttgaagct 9180gcactatggc aacaatggga ccgacaatcc ctaattatgt ttatcactgc attcttgaat 9240attgctctcc agttaccgtg tgaaagttct gctgtcgttg tttcagggtt aagaacattg 9300gttcctcaat cagataatga ggaagcttca accaacccgg ggacatgctc atggtctgat 9360gagggtaccc cttaataagg ctgactaaaa cactatataa ccttctactt gatcacaata 9420ctccgtatac ctatcatcat atatttaatc aagacgatat cctttaaaac ttattcagta 9480ctataatcac tctcgtttca aattaataag atgtgcatga ttgccctaat atatgaagag 9540gtatgataca accctaacag tggtcaaaga aaatcataat ctcgtatcgc tcgtaatata 9600acctgccaag catacctctt gcacaaagtg attcttgtac acaaataatg ttttactcta 9660caggaggtag caacgatcca tcccatcaaa aaataagtat ttcatgactt actaatgatc 9720tcttaaaata ttaagaaaaa ctgacggaac ataaattctt tatgcttcaa gctgtggagg 9780aggtgtttgg tattggctat tgttatatta caatcaataa caagcttgta aaaatattgt 9840tcttgtttca agaggtagat tgtgaccgga aatgctaaac taatgatgaa gattaatgcg 9900gaggtctgat aagaataaac cttattattc agattaggcc ccaagaggca ttcttcatct 9960ccttttagca aagtactatt tcagggtagt ccaattagtg gcacgtcttt tagctgtata 10020tcagtcgccc ctgagatacg ccacaaaagt gtctctaagc taaattggtc tgtacacatc 10080ccatacattg tattaggggc aataatatct aattgaactt agccgtttaa aatttagtgc 10140ataaatctgg gctaacacca ccaggtcaac tccattggct gaaaagaagc ttacctacaa 10200cgaacatcac tttgagcgcc ctcacaatta aaaaatagga acgtcgttcc aacaatcgag 10260cgcaaggttt caaggttgaa ctgagagtgt ctagacaaca aaatattgat actccagaca 10320ccaagcaaga cctgagaaaa aaaccatggc taaagctacg ggacgataca atctaatatc 10380gcccaaaaag gacctggaga aaggggttgt cttaagcgac ctctgtaact tcttagttag 10440ccaaactatt caggggtgga aggtttattg ggctggtatt gagtttgatg tgattcacaa 10500aggaatggcc ctattgcata gactgaaaac taatgacttt gcccctgcat ggtcaatgac 10560aaggaatctc tttcctcatt tatttcaaaa tccgaattcc acaattgaat caccgctgtg 10620ggcattgaga gtcatccttg cagcagggat acaggaccag ctgattgacc agtctttgat 10680tgaaccctta gcaggagccc ttggtctgat ctctgattgg ctgctaacaa ccaacactaa 10740ccatttcaac atgcgaacac aacgtgtcaa ggaacaattg agcctaaaaa tgctgtcgtt 10800gattcgatcc aatattctca agtttattaa caaattggat gctctacatg tcgtgaacta 10860caacggattg ttgagcagta ttgaaattgg aactcaaaat catacaatca tcataactcg 10920aactaacatg ggttttctgg tggagctcca agaacccgac aaatcggcaa tgaaccgcat 10980gaagcctggg ccggcgaaat tttccctcct tcatgagtcc acactgaaag catttacaca 11040aggatcctcg acacgaatgc aaagtttgat tcttgaattt aatagctctc ttgctatcta 11100actaaggtag aatacttcat attgagctaa ctcatatatg ctgactcaat agttatcttg 11160acatctctgc tttcataatc agatatataa gcataataaa taaatactca tatttcttga 11220taatttgttt aaccacagat aaatcctcac tgtaagccag cttccaagtt gacaccctta 11280caaaaaccag gactcagaat ccctcaaaca agagattcca agacaacatc atagaattgc 11340tttattatat gaataagcat tttatcacca gaaatcctat atactaaatg gttaattgta 11400actgaacccg caggtcacat gtgttaggtt tcacagattc tatatattac taactctata 11460ctcgtaatta acattagata agtagattaa gaaaaaagcc tgaggaagat taagaaaaac 11520tgcttattgg gtctttccgt gttttagatg aagcagttga aattcttcct cttgatatta 11580aatggctaca caacataccc aatacccaga cgctaggtta tcatcaccaa ttgtattgga 11640ccaatgtgac ctagtcacta gagcttgcgg gttatattca tcatactccc ttaatccgca 11700actacgcaac tgtaaactcc cgaaacatat ctaccgtttg aaatacgatg taactgttac 11760caagttcttg agtgatgtac cagtggcgac attgcccata gatttcatag tcccagttct 11820tctcaaggca ctgtcaggca atggattctg tcctgttgag ccgcggtgcc aacagttctt 11880agatgaaatc attaagtaca caatgcaaga tgctctcttc ttgaaatatt atctcaaaaa 11940tgtgggtgct caagaagact gtgttgatga acactttcaa gagaaaatct tatcttcaat 12000tcagggcaat gaatttttac atcaaatgtt tttctggtat gatctggcta ttttaactcg 12060aaggggtaga ttaaatcgag gaaactctag atcaacatgg tttgttcatg atgatttaat 12120agacatctta ggctatgggg actatgtttt ttggaagatc ccaatttcaa tgttaccact 12180gaacacacaa ggaatccccc atgctgctat ggactggtat caggcatcag tattcaaaga 12240agcggttcaa gggcatacac acattgtttc tgtttctact gccgacgtct tgataatgtg 12300caaagattta attacatgtc gattcaacac aactctaatc tcaaaaatag cagagattga 12360ggatccagtt tgttctgatt atcccaattt taagattgtg tctatgcttt accagagcgg 12420agattactta ctctccatat tagggtctga tgggtataaa attattaagt tcctcgaacc 12480attgtgcttg gccaaaattc aattatgctc aaagtacact gagaggaagg gccgattctt 12540aacacaaatg catttagctg taaatcacac cctagaagaa attacagaaa tgcgtgcact 12600aaagccttca caggctcaaa agatccgtga attccataga acattgataa ggctggagat 12660gacgccacaa caactttgtg agctattttc cattcaaaaa cactgggggc atcctgtgct 12720acatagtgaa acagcaatcc aaaaagttaa aaaacatgct acggtgctaa aagcattacg 12780ccctatagtg attttcgaga catactgtgt ttttaaatat agtattgcca aacattattt 12840tgatagtcaa ggatcttggt acagtgttac ttcagatagg aatctaacac cgggtcttaa 12900ttcttatatc aaaagaaatc aattccctcc gttgccaatg attaaagaac tactatggga 12960attttaccac cttgaccacc ctccactttt ctcaaccaaa attattagtg acttaagtat 13020ttttataaaa gacagagcta ccgcagtaga aaggacatgc tgggatgcag tattcgagcc 13080taatgttcta ggatataatc cacctcacaa atttagtact aaacgtgtac cggaacaatt 13140tttagagcaa gaaaactttt ctattgagaa tgttctttcc tacgcacaaa aactcgagta 13200tctactacca caatatcgga acttttcttt ctcattgaaa gagaaagagt tgaatgtagg 13260tagaaccttc ggaaaattgc cttatccgac tcgcaatgtt caaacacttt gtgaagctct 13320gttagctgat ggtcttgcta aagcatttcc tagcaatatg atggtagtta cggaacgtga 13380gcaaaaagaa agcttattgc atcaagcatc atggcaccac acaagtgatg attttggtga 13440acatgccaca gttagaggga gtagctttgt aactgattta gagaaataca atcttgcatt 13500tagatatgag tttacagcac cttttataga atattgcaac cgttgctatg gtgttaagaa 13560tgtttttaat tggatgcatt atacaatccc acagtgttat atgcatgtca gtgattatta 13620taatccacca cataacctca cactggagaa tcgagacaac ccccccgaag ggcctagttc 13680atacaggggt catatgggag ggattgaagg actgcaacaa aaactctgga caagtatttc 13740atgtgctcaa atttctttag ttgaaattaa gactggtttt aagttacgct cagctgtgat 13800gggtgacaat cagtgcatta ctgttttatc agtcttcccc ttagagactg acgcagacga 13860gcaggaacag agcgccgaag acaatgcagc gagggtggcc gccagcctag caaaagttac 13920aagtgcctgt ggaatctttt taaaacctga tgaaacattt gtacattcag gttttatcta 13980ttttggaaaa aaacaatatt tgaatggggt ccaattgcct cagtccctta aaacggctac 14040aagaatggca ccattgtctg atgcaatttt tgatgatctt caagggaccc tggctagtat 14100aggcactgct tttgagcgat ccatctctga gacacgacat atctttcctt gcaggataac 14160cgcagctttc catacgtttt tttcggtgag aatcttgcaa tatcatcatc tcgggttcaa 14220taaaggtttt gaccttggac agttaacact cggcaaacct ctggatttcg gaacaatatc 14280attggcacta gcggtaccgc aggtgcttgg agggttatcc ttcttgaatc ctgagaaatg 14340tttctaccgg aatctaggag atccagttac ctcaggctta ctccagttaa aaacttatct 14400ccgaatgatt gagatggatg atttattctt acctttaatt gcgaagaacc ctgggaactg 14460cactgccatt gactttgtgc taaatcctag cggattaaat gtccctgggt cgcaagactt 14520aacttcattt ctgcgccaga ttgtacgcag gaccatcacc ctaagtgcga aaaacaaact 14580tattaatacc ttatttcatg cgtcagctga cttcgaagac gaaatggttt gtaaatggct 14640attatcatca actcctgtta tgagtcgttt tgcggccgat atcttttcac gcacgccgag 14700cgggaagcga ttgcaaattc taggatacct ggaaggaaca cgcacattat tagcctctaa 14760gatcatcaac aataatacag agacaccggt tttggacaga ctgaggaaaa taacattgca 14820aaggtggagc ctatggttta gttatcttga tcattgtgat aatatcctgg cggaggcttt 14880aacccaaata acttgcacag ttgatttagc acagattctg agggaatatt catgggctca 14940tattttagag ggaagacctc ttattggagc cacactccca tgtatgattg agcaattcaa 15000agtgttttgg ctgaaaccct acgaacaatg tccgcagtgt tcaaatgcaa agcaaccagg 15060tgggaaacca ttcgtgtcag tggcagtcaa gaaacatatt gttagtgcat ggccgaacgc 15120atcccgaata agctggacta tcggggatgg aatcccatac attggatcaa ggacagaaga 15180taagatagga caacctgcta ttaaaccaaa atgtccttcc gcagccttaa gagaggccat 15240tgaattggcg tcccgtttaa catgggtaac tcaaggcagt tcgaacagtg acttgctaat 15300aaaaccattt ttggaagcac gagtaaattt aagtgttcaa gaaatacttc aaatgacccc 15360ttcacattac tcaggaaata ttgttcacag gtacaacgat caatacagtc ctcattcttt 15420catggccaat cgtatgagta attcagcaac gcgattgatt gtttctacaa acactttagg 15480tgagttttca ggaggtggcc agtctgcacg cgacagcaat attattttcc agaatgttat 15540aaattatgca gttgcactgt tcgatattaa atttagaaac actgaggcta cagatatcca 15600atataatcgt gctcaccttc atctaactaa gtgttgcacc cgggaagtac cagctcagta 15660tttaacatac acatctacat tggatttaga tttaacaaga taccgagaaa acgaattgat 15720ttatgacagt aatcctctaa aaggaggact caattgcaat atctcattcg ataatccatt 15780tttccaaggt aaacggctga acattataga agatgatctt attcgactgc ctcacttatc 15840tggatgggag ctagccaaga ccatcatgca atcaattatt tcagatagca acaattcatc 15900tacagaccca attagcagtg gagaaacaag atcattcact acccatttct taacttatcc 15960caagatagga cttctgtaca gttttggggc ctttgtaagt tattatcttg gcaatacaat 16020tcttcggact aagaaattaa cacttgacaa ttttttatat tacttaacta ctcaaattca 16080taatctacca catcgctcat tgcgaatact taagccaaca ttcaaacatg caagcgttat 16140gtcacggtta atgagtattg atcctcattt ttctgtttac ataggcggtg ctgcaggtga 16200cagaggactc tcagatgcgg ccaggttatt tttgagaacg tccatttcat cttttcttac 16260atttgtaaaa gaatggataa ttaatcgcgg aacaattgtc cctttatgga tagtatatcc 16320gctagagggt caaaacccaa cacctgtgaa taattttctc tatcagatcg tagaactgct 16380ggtgcatgat tcatcaagac aacaggcttt taaaactacc ataagtgatc atgtacatcc 16440tcacgacaat cttgtttaca catgtaagag tacagccagc aatttcttcc atgcatcatt 16500ggcgtactgg aggagcagac acagaaacag caaccgaaaa tacttggcaa

gagactcttc 16560aactggatca agcacaaaca acagtgatgg tcatattgag agaagtcaag aacaaaccac 16620cagagatcca catgatggca ctgaacggaa tctagtccta caaatgagcc atgaaataaa 16680aagaacgaca attccacaag aaaacacgca ccagggtccg tcgttccagt cctttctaag 16740tgactctgct tgtgggacag caaatccaaa actaaatttc gatcgatcga gacacaatgt 16800gaaatttcag gatcataact cggcatccaa gagggaaggt catcaaataa tctcacaccg 16860tctagtccta cctttcttta cattatctca agggacacgc caattaacgt catccaatga 16920gtcacaaacc caagacgaga tatcaaagta cttacggcaa ttgagatccg tcattgatac 16980cacagtttat tgtagattta ccggtatagt ctcgtccatg cattacaaac ttgatgaggt 17040cctttgggaa atagagagtt tcaagtcggc tgtgacgcta gcagagggag aaggtgctgg 17100tgccttacta ttgattcaga aataccaagt taagacctta tttttcaaca cgctagctac 17160tgagtccagt atagagtcag aaatagtatc aggaatgact actcctagga tgcttctacc 17220tgttatgtca aaattccata atgaccaaat tgagattatt cttaacaact cagcaagcca 17280aataacagac ataacaaatc ctacttggtt taaagaccaa agagcaaggc tacctaagca 17340agtcgaggtt ataaccatgg atgcagagac aacagagaat ataaacagat cgaaattgta 17400cgaagctgta tataaattga tcttacacca tattgatcct agcgtattga aagcagtggt 17460ccttaaagtc tttctaagtg atactgaggg tatgttatgg ctaaatgata atttagcccc 17520gttttttgcc actggttatt taattaagcc aataacgtca agtgctagat ctagtgagtg 17580gtatctttgt ctgacgaact tcttatcaac tacacgtaag atgccacacc aaaaccatct 17640cagttgtaaa caggtaatac ttacggcatt gcaactgcaa attcaacgaa gcccatactg 17700gctaagtcat ttaactcagt atgctgactg tgagttacat ttaagttata tccgccttgg 17760ttttccatca ttagagaaag tactatacca caggtataac ctcgtcgatt caaaaagagg 17820tccactagtc tctatcactc agcacttagc acatcttaga gcagagattc gagaattaac 17880taatgattat aatcaacagc gacaaagtcg gactcaaaca tatcacttta ttcgtactgc 17940aaaaggacga atcacaaaac tagtcaatga ttatttaaaa ttctttctta ttgtgcaagc 18000attaaaacat aatgggacat ggcaagctga gtttaagaaa ttaccagagt tgattagtgt 18060gtgcaatagg ttctaccata ttagagattg caattgtgaa gaacgtttct tagttcaaac 18120cttatattta catagaatgc aggattctga agttaagctt atcgaaaggc tgacagggct 18180tctgagttta tttccggatg gtctctacag gtttgattga attaccgtgc atagtatcct 18240gatacttgca aaggttggtt attaacatac agattataaa aaactcataa attgctctca 18300tacatcatat tgatctaatc tcaataaaca actatttaaa taacgaaagg agtccctata 18360ttatatacta tatttagcct ctctccctgc gtgataatca aaaaattcac aatgcagcat 18420gtgtgacata ttactgccgc aatgaattta acgcaacata ataaactctg cactctttat 18480aattaagctt taacgaaagg tctgggctca tattgttatt gatataataa tgttgtatca 18540atatcctgtc agatggaata gtgttttggt tgataacaca acttcttaaa acaaaattga 18600tctttaagat taagtttttt ataattatca ttactttaat ttgtcgtttt aaaaacggtg 18660atagccttaa tctttgtgta aaataagaga ttaggtgtaa taaccttaac atttttgtct 18720agtaagctac tatttcatac agaatgataa aattaaaaga aaaggcagga ctgtaaaatc 18780agaaatacct tctttacaat atagcagact agataataat cttcgtgtta atgataatta 18840agacattgac cacgctcatc agaaggctcg ccagaataaa cgttgcaaaa aggattcctg 18900gaaaaatggt cgcacacaaa aatttaaaaa taaatctatt tcttcttttt tgtgtgtcca 1896012288PRTZaire ebolavirus 12Met Glu Ala Ser Tyr Glu Arg Gly Arg Pro Arg Ala Ala Arg Gln His1 5 10 15 Ser Arg Asp Gly His Asp His His Val Arg Ala Arg Ser Ser Ser Arg 20 25 30 Glu Asn Tyr Arg Gly Glu Tyr Arg Gln Ser Arg Ser Ala Ser Gln Val 35 40 45 Arg Val Pro Thr Val Phe His Lys Lys Arg Val Glu Pro Leu Thr Val 50 55 60 Pro Pro Ala Pro Lys Asp Ile Cys Pro Thr Leu Lys Lys Gly Phe Leu65 70 75 80 Cys Asp Ser Ser Phe Cys Lys Lys Asp His Gln Leu Glu Ser Leu Thr 85 90 95 Asp Arg Glu Leu Leu Leu Leu Ile Ala Arg Lys Thr Cys Gly Ser Val 100 105 110 Glu Gln Gln Leu Asn Ile Thr Ala Pro Lys Asp Ser Arg Leu Ala Asn 115 120 125 Pro Thr Ala Asp Asp Phe Gln Gln Glu Glu Gly Pro Lys Ile Thr Leu 130 135 140 Leu Thr Leu Ile Lys Thr Ala Glu His Trp Ala Arg Gln Asp Ile Arg145 150 155 160 Thr Ile Glu Asp Ser Lys Leu Arg Ala Leu Leu Thr Leu Cys Ala Val 165 170 175 Met Thr Arg Lys Phe Ser Lys Ser Gln Leu Ser Leu Leu Cys Glu Thr 180 185 190 His Leu Arg Arg Glu Gly Leu Gly Gln Asp Gln Ala Glu Pro Val Leu 195 200 205 Glu Val Tyr Gln Arg Leu His Ser Asp Lys Gly Gly Ser Phe Glu Ala 210 215 220 Ala Leu Trp Gln Gln Trp Asp Arg Gln Ser Leu Ile Met Phe Ile Thr225 230 235 240 Ala Phe Leu Asn Ile Ala Leu Gln Leu Pro Cys Glu Ser Ser Ala Val 245 250 255 Val Val Ser Gly Leu Arg Thr Leu Val Pro Gln Ser Asp Asn Glu Glu 260 265 270 Ala Ser Thr Asn Pro Gly Thr Cys Ser Trp Ser Asp Glu Gly Thr Pro 275 280 285 1311742DNAZaire ebolavirus 13cggacacaca aaaagaaaga agaattttta ggatcttttg tgtgcgaata actatgagga 60agattaataa ttttcctctc attgaaattt atatcggaat ttaaattgaa attgttactg 120taatcacacc tggtttgttt cagagccaca tcacaaagat agagaacaac ctaggtctcc 180gaagggagca agggcatcag tgtgctcagt tgaaaatccc ttgtcaacac ctaggtctta 240tcacatcaca agttccacct cagactctgc agggtgatcc aacaacctta atagaaacat 300tattgttaaa ggacagcatt agttcacagt caaacaagca agattgagaa ttaaccttgg 360ttttgaactt gaacacttag gggattgaag attcaacaac cctaaagctt ggggtaaaac 420attggaaata gttaaaagac aaattgctcg gaatcacaaa attccgagta tggattctcg 480tcctcagaaa atctggatgg cgccgagtct cactgaatct gacatggatt accacaagat 540cttgacagca ggtctgtccg ttcaacaggg gattgttcgg caaagagtca tcccagtgta 600tcaagtaaac aatcttgaag aaatttgcca acttatcata caggcctttg aagcaggtgt 660tgattttcaa gagagtgcgg acagtttcct tctcatgctt tgtcttcatc atgcgtacca 720gggagattac aaacttttct tggaaagtgg cgcagtcaag tatttggaag ggcacgggtt 780ccgttttgaa gtcaagaagc gtgatggagt gaagcgcctt gaggaattgc tgccagcagt 840atctagtgga aaaaacatta agagaacact tgctgccatg ccggaagagg agacaactga 900agctaatgcc ggtcagtttc tctcctttgc aagtctattc cttccgaaat tggtagtagg 960agaaaaggct tgccttgaga aggttcaaag gcaaattcaa gtacatgcag agcaaggact 1020gatacaatat ccaacagctt ggcaatcagt aggacacatg atggtgattt tccgtttgat 1080gcgaacaaat tttctgatca aatttctcct aatacaccaa gggatgcaca tggttgccgg 1140gcatgatgcc aacgatgctg tgatttcaaa ttcagtggct caagctcgtt tttcaggctt 1200attgattgtc aaaacagtac ttgatcatat cctacaaaag acagaacgag gagttcgtct 1260ccatcctctt gcaaggaccg ccaaggtaaa aaatgaggtg aactccttta aggctgcact 1320cagctccctg gccaagcatg gagagtatgc tcctttcgcc cgacttttga acctttctgg 1380agtaaataat cttgagcatg gtcttttccc tcaactatcg gcaattgcac tcggagtcgc 1440cacagcacac gggagtaccc tcgcaggagt aaatgttgga gaacagtatc aacaactcag 1500agaggctgcc actgaggctg agaagcaact ccaacaatat gcagagtctc gcgaacttga 1560ccatcttgga cttgatgatc aggaaaagaa aattcttatg aacttccatc agaaaaagaa 1620cgaaatcagc ttccagcaaa caaacgctat ggtaactcta agaaaagagc gcctggccaa 1680gctgacagaa gctatcactg ctgcgtcact gcccaaaaca agtggacatt acgatgatga 1740tgacgacatt ccctttccag gacccatcaa tgatgacgac aatcctggcc atcaagatga 1800tgatccgact gactcacagg atacgaccat tcccgatgtg gtggttgatc ccgatgatgg 1860aagctacggc gaataccaga gttactcgga aaacggcatg aatgcaccag atgacttggt 1920cctattcgat ctagacgagg acgacgagga cactaagcca gtgcctaata gatcgaccaa 1980gggtggacaa cagaagaaca gtcaaaaggg ccagcatata gagggcagac agacacaatc 2040caggccaatt caaaatgtcc caggccctca cagaacaatc caccacgcca gtgcgccact 2100cacggacaat gacagaagaa atgaaccctc cggctcaacc agccctcgca tgctgacacc 2160aattaacgaa gaggcagacc cactggacga tgccgacgac gagacgtcta gccttccgcc 2220cttggagtca gatgatgaag agcaggacag ggacggaact tccaaccgca cacccactgt 2280cgccccaccg gctcccgtat acagagatca ctctgaaaag aaagaactcc cgcaagacga 2340gcaacaagat caggaccaca ctcaagaggc caggaaccag gacagtgaca acacccagtc 2400agaacactct tttgaggaga tgtatcgcca cattctaaga tcacaggggc catttgatgc 2460tgttttgtat tatcatatga tgaaggatga gcctgtagtt ttcagtacca gtgatggcaa 2520agagtacacg tatccagact cccttgaaga ggaatatcca ccatggctca ctgaaaaaga 2580ggctatgaat gaagagaata gatttgttac attggatggt caacaatttt attggccggt 2640gatgaatcac aagaataaat tcatggcaat cctgcaacat catcagtgaa tgagcatgga 2700acaatgggat gattcaaccg acaaatagct aacattaagt agtcaaggaa cgaaaacagg 2760aagaattttt gatgtctaag gtgtgaatta ttatcacaat aaaagtgatt cttatttttg 2820aatttaaagc tagccttatt attactagcc gtttttcaaa gttcaatttg agtcttaatg 2880caaataggcg ttaagccaca gttatagcca taattgtaac tcaatattct aactagcgat 2940ttatctaaat taaattacat tatgctttta taacttacct actagcctgc ccaacattta 3000cacgatcgtt ttataattaa gaaaaaacta atgatgaaga ttaaaacctt catcatcctt 3060acgtcaattg aattctctag cactcgaagc ttattgtctt caatgtaaaa gaaaagctgg 3120tctaacaaga tgacaactag aacaaagggc aggggccata ctgcggccac gactcaaaac 3180gacagaatgc caggccctga gctttcgggc tggatctctg agcagctaat gaccggaaga 3240attcctgtaa gcgacatctt ctgtgatatt gagaacaatc caggattatg ctacgcatcc 3300caaatgcaac aaacgaagcc aaacccgaag acgcgcaaca gtcaaaccca aacggaccca 3360atttgcaatc atagttttga ggaggtagta caaacattgg cttcattggc tactgttgtg 3420caacaacaaa ccatcgcatc agaatcatta gaacaacgca ttacgagtct tgagaatggt 3480ctaaagccag tttatgatat ggcaaaaaca atctcctcat tgaacagggt ttgtgctgag 3540atggttgcaa aatatgatct tctggtgatg acaaccggtc gggcaacagc aaccgctgcg 3600gcaactgagg cttattgggc cgaacatggt caaccaccac ctggaccatc actttatgaa 3660gaaagtgcga ttcggggtaa gattgaatct agagatgaga ccgtccctca aagtgttagg 3720gaggcattca acaatctaaa cagtaccact tcactaactg aggaaaattt tgggaaacct 3780gacatttcgg caaaggattt gagaaacatt atgtatgatc acttgcctgg ttttggaact 3840gctttccacc aattagtaca agtgatttgt aaattgggaa aagatagcaa ctcattggac 3900atcattcatg ctgagttcca ggccagcctg gctgaaggag actctcctca atgtgcccta 3960attcaaatta caaaaagagt tccaatcttc caagatgctg ctccacctgt catccacatc 4020cgctctcgag gtgacattcc ccgagcttgc cagaaaagct tgcgtccagt cccaccatcg 4080cccaagattg atcgaggttg ggtatgtgtt tttcagcttc aagatggtaa aacacttgga 4140ctcaaaattt gagccaatct cccttccctc cgaaagaggc gaataatagc agaggcttca 4200actgctgaac tatagggtac gttacattaa tgatacactt gtgagtatca gccctggata 4260atataagtca attaaacgac caagataaaa ttgttcatat ctcgctagca gcttaaaata 4320taaatgtaat aggagctata tctctgacag tattataatc aattgttatt aagtaaccca 4380aaccaaaagt gatgaagatt aagaaaaacc tacctcggct gagagagtgt tttttcatta 4440accttcatct tgtaaacgtt gagcaaaatt gttaaaaata tgaggcgggt tatattgcct 4500actgctcctc ctgaatatat ggaggccata taccctgtca ggtcaaattc aacaattgct 4560agaggtggca acagcaatac aggcttcctg acaccggagt cagtcaatgg ggacactcca 4620tcgaatccac tcaggccaat tgccgatgac accatcgacc atgccagcca cacaccaggc 4680agtgtgtcat cagcattcat ccttgaagct atggtgaatg tcatatcggg ccccaaagtg 4740ctaatgaagc aaattccaat ttggcttcct ctaggtgtcg ctgatcaaaa gacctacagc 4800tttgactcaa ctacggccgc catcatgctt gcttcataca ctatcaccca tttcggcaag 4860gcaaccaatc cacttgtcag agtcaatcgg ctgggtcctg gaatcccgga tcatcccctc 4920aggctcctgc gaattggaaa ccaggctttc ctccaggagt tcgttcttcc gccagtccaa 4980ctaccccagt atttcacctt tgatttgaca gcactcaaac tgatcaccca accactgcct 5040gctgcaacat ggaccgatga cactccaaca ggatcaaatg gggcgttgcg tccaggaatt 5100tcatttcatc caaaacttcg ccccattctt ttacccaaca aaagtgggaa gaaggggaac 5160agtgccgatc taacatctcc ggagaaaatc caagcaataa tgacttcact ccaggacttt 5220aagatcgttc caattgatcc aaccaaaaat atcatgggaa tcgaagtgcc agaaactctg 5280gtccacaagc tgaccggtaa gaaggtgact tctaaaaatg gacaaccaat catccctgtt 5340cttttgccaa agtacattgg gttggacccg gtggctccag gagacctcac catggtaatc 5400acacaggatt gtgacacgtg tcattctcct gcaagtcttc cagctgtgat tgagaagtaa 5460ttgcaataat tgactcagat ccagttttat agaatcttct cagggatagt gcataacatc 5520tatttagtaa tccgtccatt agaggagaca cttttaattg atcaatatac taaaggtgct 5580ttacaccatt gtcttttttc tctcctaaat gtagaactta acaaaagact cataatatac 5640ttgtttttaa aggattgatt gatgaaagat cataactaat aacattacaa ataatcctac 5700tataatcaat acggtgattc aaatgttaat ctttctcatt gcacatactt tttgccctta 5760tcctcaaatt gcctgcatgc ttacatctga ggatagccag tgtgacttgg attggaaatg 5820tggagaaaaa atcgggaccc atttctaggt tgttcacaat ccaagtacag acattgccct 5880tctaattaag aaaaaatcgg cgatgaagat taagccgaca gtgagcgtaa tcttcatctc 5940tcttagatta tttgttttcc agagtagggg tcgtcaggtc cttttcaatc gtgtaaccaa 6000aataaactcc actagaagga tattgtgggg caacaacaca atgggcgtta caggaatatt 6060gcagttacct cgtgatcgat tcaagaggac atcattcttt ctttgggtaa ttatcctttt 6120ccaaagaaca ttttccatcc cacttggagt catccacaat agcacattac aggttagtga 6180tgtcgacaaa ctagtttgtc gtgacaaact gtcatccaca aatcaattga gatcagttgg 6240actgaatctc gaagggaatg gagtggcaac tgacgtgcca tctgcaacta aaagatgggg 6300cttcaggtcc ggtgtcccac caaaggtggt caattatgaa gctggtgaat gggctgaaaa 6360ctgctacaat cttgaaatca aaaaacctga cgggagtgag tgtctaccag cagcgccaga 6420cgggattcgg ggcttccccc ggtgccggta tgtgcacaaa gtatcaggaa cgggaccgtg 6480tgccggagac tttgccttcc ataaagaggg tgctttcttc ctgtatgatc gacttgcttc 6540cacagttatc taccgaggaa cgactttcgc tgaaggtgtc gttgcatttc tgatactgcc 6600ccaagctaag aaggacttct tcagctcaca ccccttgaga gagccggtca atgcaacgga 6660ggacccgtct agtggctact attctaccac aattagatat caggctaccg gttttggaac 6720caatgagaca gagtacttgt tcgaggttga caatttgacc tacgtccaac ttgaatcaag 6780attcacacca cagtttctgc tccagctgaa tgagacaata tatacaagtg ggaaaaggag 6840caataccacg ggaaaactaa tttggaaggt caaccccgaa attgatacaa caatcgggga 6900gtgggccttc tgggaaacta aaaaaaacct cactagaaaa attcgcagtg aagagttgtc 6960tttcacagtt gtatcaaacg gagccaaaaa catcagtggt cagagtccgg cgcgaacttc 7020ttccgaccca gggaccaaca caacaactga agaccacaaa atcatggctt cagaaaattc 7080ctctgcaatg gttcaagtgc acagtcaagg aagggaagct gcagtgtcgc atctaacaac 7140ccttgccaca atctccacga gtccccaatc cctcacaacc aaaccaggtc cggacaacag 7200cacccataat acacccgtgt ataaacttga catctctgag gcaactcaag ttgaacaaca 7260tcaccgcaga acagacaacg acagcacagc ctccgacact ccctctgcca cgaccgcagc 7320cggaccccca aaagcagaga acaccaacac gagcaagagc actgacttcc tggaccccgc 7380caccacaaca agtccccaaa accacagcga gaccgctggc aacaacaaca ctcatcacca 7440agataccgga gaagagagtg ccagcagcgg gaagctaggc ttaattacca atactattgc 7500tggagtcgca ggactgatca caggcgggag aagaactcga agagaagcaa ttgtcaatgc 7560tcaacccaaa tgcaacccta atttacatta ctggactact caggatgaag gtgctgcaat 7620cggactggcc tggataccat atttcgggcc agcagccgag ggaatttaca tagaggggct 7680aatgcacaat caagatggtt taatctgtgg gttgagacag ctggccaacg agacgactca 7740agctcttcaa ctgttcctga gagccacaac tgagctacgc accttttcaa tcctcaaccg 7800taaggcaatt gatttcttgc tgcagcgatg gggcggcaca tgccacattc tgggaccgga 7860ctgctgtatc gaaccacatg attggaccaa gaacataaca gacaaaattg atcagattat 7920tcatgatttt gttgataaaa cccttccgga ccagggggac aatgacaatt ggtggacagg 7980atggagacaa tggataccgg caggtattgg agttacaggc gttataattg cagttatcgc 8040tttattctgt atatgcaaat ttgtctttta gtttttcttc agattgcttc atggaaaagc 8100tcagcctcaa atcaatgaaa ccaggattta attatatgga ttacttgaat ctaagattac 8160ttgacaaatg ataatataat acactggagc tttaaacata gccaatgtga ttctaactcc 8220tttaaactca cagttaatca taaacaaggt ttgacatcaa tctagttatc tctttgagaa 8280tgataaactt gatgaagatt aagaaaaagg taatctttcg attatcttta atcttcatcc 8340ttgattctac aatcatgaca gttgtcttta gtgacaaggg aaagaagcct ttttattaag 8400ttgtaataat cagatctgcg aaccggtaga gtttagttgc aacctaacac acataaagca 8460ttggtcaaaa gtcaatagaa atttaaacag tgagtggaga caacttttaa atggaagctt 8520catatgagag aggacgccca cgagctgcca gacagcattc aagggatgga cacgaccacc 8580atgttcgagc acgatcatca tccagagaga attatcgagg tgagtaccgt caatcaagga 8640gcgcctcaca agtgcgcgtt cctactgtat ttcataagaa gagagttgaa ccattaacag 8700ttcctccagc acctaaagac atatgtccga ccttgaaaaa aggatttttg tgtgacagta 8760gtttttgcaa aaaagatcac cagttggaga gtttaactga tagggaatta ctcctactaa 8820tcgcccgtaa gacttgtgga tcagtagaac aacaattaaa tataactgca cccaaggact 8880cgcgcttagc aaatccaacg gctgatgatt tccagcaaga ggaaggtcca aaaattacct 8940tgttgacact gatcaagacg gcagaacact gggcgagaca agacatcaga accatagagg 9000attcaaaatt aagagcattg ttgactctat gtgctgtgat gacgaggaaa ttctcaaaat 9060cccagctgag tcttttatgt gagacacacc taaggcgcga ggggcttggg caagatcagg 9120cagaacccgt tctcgaagta tatcaacgat tacacagtga taaaggaggc agttttgaag 9180ctgcactatg gcaacaatgg gaccgacaat ccctaattat gtttatcact gcattcttga 9240atattgctct ccagttaccg tgtgaaagtt ctgctgcgtt gtttcagggt taagaacatt 9300ggttcctcaa tcagataatg aggaagcttc aaccaacccg gggacatgct catggtctga 9360tgagggtacc ccttaataag gctgactaaa acactatata accttctact tgatcacaat 9420actccgtata cctatcatca tatatttaat caagacgata tcctttaaaa cttattcagt 9480actataatca ctctcgtttc aaattaataa gatgagcatg attgccctaa tatatgaaga 9540ggtatgatac aaccctaaca gtgatcaaag aaaatcataa tctcgtatcg ctcgtaatat 9600aacctgccaa gcatacctct tgcacaaagt gattcttgtc cacaaataat gttttactct 9660acaggaggta gcaacgatcc atcccatcaa aaaataagta tttcatgact tactaatgat 9720ctcttaaaat attaagaaaa actgacggaa cataaattct ttatgcttca agctgtggag 9780gaggtgtttg gtattggcta ttgttatatt acaatcaata acaagcttgt aaaaatattg 9840ttcttgtttc aagaggtaga ttgtgaccgg aaatgctaaa ctaatgatga agattaatgc 9900ggaggtctga taagaataaa ccttattatt cagattaggc cccaagaggc attcttcatc 9960tccttttagc aaagtactat ttcagggtag tccaattagt ggcacgtctt ttagctgtat 10020atcagtcgcc cctgagatac gccacaaaag tgtctctaag ctaaattggt ctgtacacat 10080cccatacatt gtattagggg caataatatc taattgaact tagccgttta aaatttagtg 10140cataaatctg ggctaacacc accaggtcaa ctccattggc tgaaaagaag cttacctaca 10200acgaacatca ctttgagcgc cctcacaatt aaaaaatagg aacgtcgttc caacaatcga 10260gcgcaaggtt tcaaggttga actgagagtg tctagacaac aaaatattga tactccagac 10320accaagcaag acctgagaaa aaaacatggc taaagctacg ggacgataca atctaatatc 10380gcccaaaaag gacctggaga aaggggttgt cttaagcgac ctctgtaact tcttagttag 10440ccaaactatt caggggtgga aggtttattg ggctggtatt gagtttgatg tgactcacaa 10500aggaatggcc ctattgcata gactgaaaac taatgacttt gcccctgcat ggtcaatgac 10560aaggaatctc tttcctcatt tatttcaaaa tccgaattcc acaattgaat caccgctgtg 10620ggcattgaga gtcatccttg cagcagggat acaggaccag ctgattgacc agtctttgat 10680tgaaccctta gcaggagccc ttggtctgat ctctgattgg

ctgctaacaa ccaacactaa 10740ccatttcaac atgcgaacac aacgtgtcaa ggaacaattg agcctaaaaa tgctgtcgtt 10800gattcgatcc aatattctca agtttattaa caaattggat gctctacatg tcgtgaacta 10860caacggattg ttgagcagta ttgaaattgg aactcaaaat catacaatca tcataactcg 10920aactaacatg ggttttctgg tggagctcca agaacccgac aaatcggcaa tgaaccgcat 10980gaagcctggg ccggcgaaat tttccctcct tcatgagtcc acactgaaag catttacaca 11040aggatcctcg acacgaatgc aaagtttgat tcttgaattt aatagctctc ttgctatcta 11100actaaggtag aatacttcat attgagctaa ctcatatatg ctgactcaat agttatcttg 11160acatctctgc tttcataatc agatatataa gcataataaa taaatactca tatttcttga 11220taatttgttt aaccacagat aaatcctcac tgtaagccag cttccaagtt gacaccctta 11280caaaaccagg actcagaatc cctcaaacaa gagattccaa gacaacatca tagaattgct 11340ttattatatg aataagcatt ttatcaccag aaatcctata tactaaatgg ttaattgtaa 11400ctgaacccgc aggtcacatg tgttaggttt cacagattct atatattact aactctatac 11460tcgtaattaa cattagataa gtagattaag aaaaaagcct gaggaagatt aagaaaaact 11520gcttattggg tctttccgtg ttttagatga agcagttgaa attcttcctc ttgatattaa 11580atggctacac aacataccca atacccagac gctaggttat catcaccaat tgtattggac 11640caatgtgacc tagtcactag agcttgcggg ttatattcat catactccct taatccgcaa 11700ctacgcaact gtaaactccc gaaacatatc taccgtttga aa 1174214260PRTZaire ebolavirus 14Met Glu Ala Ser Tyr Glu Arg Gly Arg Pro Arg Ala Ala Arg Gln His1 5 10 15 Ser Arg Asp Gly His Asp His His Val Arg Ala Arg Ser Ser Ser Arg 20 25 30 Glu Asn Tyr Arg Gly Glu Tyr Arg Gln Ser Arg Ser Ala Ser Gln Val 35 40 45 Arg Val Pro Thr Val Phe His Lys Lys Arg Val Glu Pro Leu Thr Val 50 55 60 Pro Pro Ala Pro Lys Asp Ile Cys Pro Thr Leu Lys Lys Gly Phe Leu65 70 75 80 Cys Asp Ser Ser Phe Cys Lys Lys Asp His Gln Leu Glu Ser Leu Thr 85 90 95 Asp Arg Glu Leu Leu Leu Leu Ile Ala Arg Lys Thr Cys Gly Ser Val 100 105 110 Glu Gln Gln Leu Asn Ile Thr Ala Pro Lys Asp Ser Arg Leu Ala Asn 115 120 125 Pro Thr Ala Asp Asp Phe Gln Gln Glu Glu Gly Pro Lys Ile Thr Leu 130 135 140 Leu Thr Leu Ile Lys Thr Ala Glu His Trp Ala Arg Gln Asp Ile Arg145 150 155 160 Thr Ile Glu Asp Ser Lys Leu Arg Ala Leu Leu Thr Leu Cys Ala Val 165 170 175 Met Thr Arg Lys Phe Ser Lys Ser Gln Leu Ser Leu Leu Cys Glu Thr 180 185 190 His Leu Arg Arg Glu Gly Leu Gly Gln Asp Gln Ala Glu Pro Val Leu 195 200 205 Glu Val Tyr Gln Arg Leu His Ser Asp Lys Gly Gly Ser Phe Glu Ala 210 215 220 Ala Leu Trp Gln Gln Trp Asp Arg Gln Ser Leu Ile Met Phe Ile Thr225 230 235 240 Ala Phe Leu Asn Ile Ala Leu Gln Leu Pro Cys Glu Ser Ser Ala Ala 245 250 255 Leu Phe Gln Gly 260 1519111DNALake Victoria marburgvirus 15agacacacaa aaacaagaga tgatgatttt gtgtatcata taaataaaga agaatattaa 60cattgacatt gagacttgtc agtctgttaa tattcttgaa gagatggatt tacacagttt 120gttggagttg ggtacaaaac ccactgcccc tcatgttcgt aataagaaag tgatattatt 180tgacacaaat catcaggtta gtatctgtaa tcagataata gatgcaataa actcagggat 240tgatcttgga gatctcctag aagggggttt gctgacgttg tgtgttgagc attactataa 300ttctgataag gataaattca acacaagtcc tatcgcgaag tacttacgtg atgcgggcta 360tgaatttgat gtcatcaaga atgcagatgc aacccgcttt ctggatgtga ttcctaatga 420acctcattac agccctttaa ttctagccct taagacattg gaaagtactg aatctcagag 480ggggagaatt gggctctttt tatcattttg cagtcttttc ctcccaaaac ttgtcgtcgg 540agaccgagct agtatcgaaa aggctttaag acaagtaaca gtgcatcaag aacaggggat 600cgtcacatac cctaatcatt ggcttaccac aggccacatg aaagtaattt tcgggatttt 660gaggtccagc ttcattttaa agtttgtgtt gattcatcaa ggagtaaatt tggtgacagg 720tcatgatgcc tatgacagta tcattagtaa ttcagtaggt caaactagat tctcaggact 780tcttatcgtg aaaacagttc tcgagttcat cttgcaaaaa actgattcag gggtgacact 840acatcctttg gtgcggacct ccaaagtaaa aaatgaagtt gctagtttca agcaggcgtt 900gagcaaccta gcccgacatg gggaatacgc accatttgca cgggttctga atttatcagg 960gattaacaac ctcgaacatg gactctatcc tcagctttca gcaattgcgc tgggtgtggc 1020aacagcacac ggcagtacat tggctggtgt caatgttggc gaacaatatc aacaactacg 1080agaggcggca catgatgcgg aagtaaaact acaaaggcga catgaacatc aggaaattca 1140agctattgcc gaggatgacg aggaaaggaa gatattagaa caattccacc ttcagaaaac 1200tgaaatcaca cacagtcaga cactagccgt cctcagccag aaacgagaaa aattagctcg 1260tctcgctgca gaaattgaaa acaatattgt ggaagatcag ggatttaagc aatcacagaa 1320tcgggtgtca cagtcgtttt tgaatgaccc tacacctgtg gaagtaacgg ttcaagccag 1380gcccatgaat cgaccaactg ctctgcctcc cccagttgac gacaagattg agcatgaatc 1440tacagaagat agctcttctt caagtagctt tgttgacttg aatgatccat ttgcactgct 1500gaatgaggac gaggatactc ttgatgacag tgtcatgatc ccgggcacaa catcgagaga 1560atttcaaggg attcctgaac cgccaagaca atcccaagac ctcaataaca gccaaggaaa 1620gcaggaagat gaatccacaa atccgattaa gaaacagttt ctgagatatc aagaattgcc 1680tcctgttcaa gaggatgatg aatcggaata cacaactgac tctcaagaaa gcatcgacca 1740accaggatcc gacaatgaac aaggagttga tcttccacct cctccgttgt acgctcagga 1800aaaaagacag gacccaatac agcacccagc agcaaaccct caggatccct tcggcagtat 1860tggtgatgta aatggtgata tcttagaacc tataagatca ccttcttcac catctgctcc 1920tcaggaagac acaaggatga gggaagccta tgaattgtcg cctgatttca caaatgatga 1980ggataatcag cagaattggc cacaaagagt ggtgacaaag aagggtagaa ctttccttta 2040tcctaatgat cttctgcaaa caaatcctcc agagtcactt ataacagccc tcgttgagga 2100ataccaaaat cctgtctcag ctaaggagct tcaagcagat tggcccgaca tgtcatttga 2160tgaaaggaga catgttgcga tgaacttgta gtccagataa cacagcacgg ttactcactt 2220atctattttg atatgactca tcctcagatc acagcaatca aatttatttg aatatttgaa 2280ccacctttta gtatcctatt acttgttact attgtgtgag acaacataag ccatcaaata 2340acaatcacgg gcaaggactg ggcatactat ggtggtctta gagcattgtc cagtgctaca 2400aattcttttt tcaattgcta taattataca actacaaacc tccatacatt tgccgcaaca 2460ctgtaatcaa cactgctgta tctcttcttc aagccatctg atttaactta ataaacatga 2520cttgattcaa agaatatact gacaatgtta ctgtttgaat ttctcaagtg gtgcactatc 2580ctactgtttt gctcagctta gtatattgta atatgtaagt ggactctccc cttctcctct 2640cgtgtattct ttataaatca cttacttgat agaatgtcga gtctactggt ttggagtttc 2700cttactctaa tggatgtaat aattaactgt tggcctagat gataacagat atgaggttat 2760ataattactc atagtgtaaa gtataattct tacctctgtt tcttctgttt tccctttctt 2820ttataatatg ccaattaaga aaaactaaaa atcgaagaat attaaagatt ttctttaata 2880ttcagaaaag gctttttatt ctattctttc tttttacaaa cgtattgaaa tagtaattct 2940cacaatgtgg gactcatcat acatgcagca agtcagcgaa gggttgatga ctggaaaagt 3000acccatagat caagtgtttg gtgccaatcc cttagagaag ttatacaaga gaagaaaacc 3060aaaaggcaca gttggactac aatgtagccc ttgtctaatg tcaaaggcga caagtactga 3120tgatattatt tgggaccaac tgatcgtgaa gagaacacta gctgatctac ttataccgat 3180aaataggcag atatcagaca ttcaaagcac tctaagcgaa gtaacaacaa gagtccatga 3240aattgagcgg caattacatg agattacccc agttttaaaa atgggaagga cactggaagc 3300aatttccaag gggatgtcag aaatgttagc caaatacgac caccttgtaa tttcaactgg 3360aagaaccact gcaccagctg ctgcctttga tgcctactta aatgagcatg gtgtccctcc 3420ccctcaaccc gcgattttca aagatcttgg ggttgcccaa caagcttgta gtaaggggac 3480catggttaaa aatgcaacaa cagatgcagc cgacaagatg tcgaaggttc ttgaactcag 3540tgaggaaacg ttctccaagc caaacctttc agctaaggat ttagcccttt tattgtttac 3600ccatctaccc ggcaacaaca ctccattcca tatcctagct caggtccttt caaaaattgc 3660ttacaagtca ggaaaatccg gagcattctt ggatgcattt caccagattc taagtgaagg 3720agagaatgct caggcggcat taactcgact aagcagaaca tttgacgctt tccttggagt 3780ggttcctcca gtgataagag tcaaaaactt ccaaacagtc cctcgtccat gtcaaaaaag 3840tcttcgggct gtccctccaa atccaacaat tgacaaagga tgggtctgtg tttattcatc 3900tgagcaaggt gaaacacggg cccttaaaat ctaattctca ttgttcatag ttgcaaggga 3960agtgatcttt ccgagttgat acaaagacac taaacatttc aaaagcatgt atgtggacaa 4020aacataatta gaccatctta attggagtag taatttattt ctgtcttaaa tgtgattttc 4080actttaaaag cgttaaatgg tgatagatta atccttgaag ttactcttct atatattata 4140gagaaaccaa tgttactaac aaaaggggtc tacctaacgc atatgattga gtaatccgta 4200tattttataa accaaacaat taacttctta ctttttaaga atcaactaac aacatagaaa 4260agacatttat ccttatgtaa tcctcggctt agttgaaatt aacttttgtt ggacctcaag 4320acgcttattc atagtatatt atatgatttt ttataagttt aagatatctt aaattatacc 4380cacaaaagat actgttttaa ttaagaaaaa ctatgaagaa cattaagaag atctttcttt 4440cgtagtgttc ttttactgga aggagtattc caatttcagc ttgttggatt aattgttact 4500taaattgtcc tttttgaaat taattcacac aaggtagttt aaatttatat ccaaaataaa 4560ttttgatatg gccagttcca gcaattacaa cacatacatg caatacttga acccccctcc 4620ttatgctgat cacggtgcaa accagttgat cccggcggat cagctatcaa atcagcaggg 4680tataactcca aattacgtgg gtgatttaaa cctagatgat cagttcaaag ggaatgtctg 4740ccatgctttc actttagagg caataattga catatctgca tataacgagc gaacagtcaa 4800aggcgttccg gcatggctgc ctcttgggat tatgagcaat tttgaatatc ctttagctca 4860tactgtggcc gcgttgctca caggcagcta tacaatcacc caatttactc acaacgggca 4920aaaattcgtc cgtgttaatc gacttggtac aggaatccca gcacacccac tcagaatgtt 4980gcgtgaagga aatcaagctt ttattcagaa tatggtgatc cccaggaatt tttcaactaa 5040tcaattcacc tacaatctca ctaatttagt attgagtgtg caaaaacttc ctgatgatgc 5100ctggcgccca tccaaggaca aattaattgg gaacactatg catcccgcag tctccatcca 5160cccgaatctg ccgcctattg ttctaccaac agtcaagaag caggcttatc gtcagcacaa 5220aaatcccaac aatggaccat tgctggccat atctggcatc ctccatcaac tgagggtcga 5280aaaagtccca gagaagacga gcctgtttag gatctcgctt cctgccgaca tgttctcagt 5340aaaagagggt atgatgaaga aaaggggaga aaattccccc gtggtttatt ttcaagcacc 5400tgagaacttc cctttgaatg gcttcaataa cagacaagtt gtgctagcgt atgcgaatcc 5460aacgctcagt gccgtttgaa atgatgctca aatgagacag gagtccatct gtataagaag 5520tatggcttaa atggatattt gtcaaattct tacaagatta gtttgtattg atttcaacaa 5580tgctttaacc ttacattgct gctttaaata gttgattaag ctgatcagct tgtaatatgt 5640aatctcttct gggccatcag atccataatg ggtttactag actatataag agaaatagta 5700atattttata aacaattctt gctcagtttt actgtgattt aataacatat gtcattgtgc 5760cctccattgc taagtcaact caactgacga taatactcct tctgaaatag taagaaaaac 5820taatgaagaa cattaattgc tgggtaaaag tgattaattt ctttaaattt gaccagaata 5880atattttgtc agtgaatata ttctcatatc acttgattaa aaacagaaaa ttaccctaac 5940atgaagacca catgtttcct tatcagtctt atcttaattc aagggacaaa aaatctcccc 6000attttagaga tagctagtaa taatcaaccc caaaatgtgg attcggtatg ctccggaact 6060ctccagaaga cagaagacgt ccatctgatg ggattcacac tgagtgggca aaaagttgct 6120gattcccctt tggaggcatc caagcgatgg gctttcagga caggtgtacc tcccaagaat 6180gttgagtaca cagaggggga ggaagccaaa acatgctaca atataagtgt aacggatccc 6240tctggaaaat ccttgctgtt agatcctcct accaacatcc gtgactatcc taaatgcaaa 6300actatccatc atattcaagg tcaaaaccct catgcacagg ggatcgccct tcatttatgg 6360ggagcatttt ttctgtatga tcgcattgcc tccacaacaa tgtaccgagg caaagtcttc 6420actgaaggga acatagcagc tatgattgtc aataagacag tgcacaaaat gattttctcg 6480cggcaaggac aagggtaccg tcatatgaat ctgacttcta ctaataaata ttggacaagt 6540agtaacggaa cgcaaacgaa tgacactgga tgtttcggcg ctcttcaaga atacaattct 6600acaaagaacc aaacatgtgc tccgtccaaa atacctccac cactgcccac agcccgtccg 6660gagatcaaac tcacaagcac cccaactgat gccaccaaac tcaataccac ggacccaagc 6720agtgatgatg aggacctcgc aacatccggc tcagggtccg gagaacgaga accccacaca 6780acttctgatg cggtcaccaa gcaagggctt tcatcaacaa tgccacccac tccctcacca 6840caaccaagca cgccacagca aggaggaaac aacacaaacc attcccaaga tgctgtgact 6900gaactagaca aaaataacac aactgcacaa ccgtccatgc cccctcataa cactaccaca 6960atctctacta acaacacctc caaacacaac ttcagcactc tctctgcacc attacaaaac 7020accaccaatg acaacacaca gagcacaatc actgaaaatg agcaaaccag tgccccctcg 7080ataacaaccc tgcctccaac gggaaatccc accacagcaa agagcaccag cagcaaaaaa 7140ggccccgcca caacggcacc aaacacgaca aatgagcatt tcaccagtcc tccccccacc 7200cccagctcga ctgcacaaca tcttgtatat ttcagaagaa agcgaagtat cctctggagg 7260gaaggcgaca tgttcccttt tctggatggg ttaataaatg ctccaattga ttttgaccca 7320gttccaaata caaaaacaat ctttgatgaa tcctctagtt ctggtgcctc ggctgaggaa 7380gatcaacatg cctcccccaa tattagttta actttatctt attttcctaa tataaatgag 7440aacactgcct actctggaga aaatgagaat gattgtgatg cagagttaag aatttggagc 7500gttcaggagg atgacctggc cgcagggctc agttggatac cgttttttgg ccctggaatt 7560gaaggacttt acactgctgt tttaattaaa aatcaaaaca atttggtctg caggttgagg 7620cgtctagcca atcaaactgc caaatccttg gaactcttat tgagagtcac aactgaggaa 7680agaacattct ccttaatcaa tagacatgct attgactttc tactcacaag atggggagga 7740acatgcaaag tgcttggacc tgattgttgc atcgggatag aagacttgtc caaaaatatt 7800tcagagcaaa ttgaccaaat taaaaaggac gaacaaaaag aggggactgg ttggggtctg 7860ggtggtaaat ggtggacatc cgactggggt gttcttacta acttgggcat tttgctacta 7920ttatccatag ctgtcttgat tgctctatcc tgtatttgtc gtatctttac taaatatatc 7980ggataacgtt aaatgtgtaa tgattaggac tttaggacaa ttgctactga gccctttttc 8040taatctactg aaatcaactt gggagatttt taagaagctg ataacttaat gtgaatcaat 8100agtttatgta ttatcgatta ttatggtttg atattcaatt gttattattg tcaggagtga 8160ccttttctat ttgatgcatt aatgttttaa actacctctt aagcctttga gggcggtccc 8220aatatgtgcg taggggttaa tttaaaggga tttcttattg tacagttttc tgtattactt 8280atttgggctt gaagacatag ttaagatttg ccgaaaatgc tctccagtca attccatccc 8340ctctcagaaa agacgtgctg ttcaaagagt cttaatttat aaccaactat tgcaagaatt 8400aatttacttt ttccgttata cttagttaca ttaatctttt gactgttcag cattattaac 8460gacttgtctt aattcaatcg ttcggatgaa attcataagg aaaaatgagc ctccttcccc 8520ctattctggg ctgagaaaat ttctcttatc cgcctaaaat ctgatctgtt aggtcatggg 8580tccttcataa tctgtttgag catgaatatt gatcaaatga ccaaatgata gtgcatttgt 8640atagactcaa ttatccttta ttaagaaaaa gataaataga acacaaagaa ttgacaaaat 8700tttactttga ttgattttgc aaggagttat aaaaatcttg aaggataaat tgttataaag 8760tagagtcgaa gaacattaaa tgttctttgt tagaattatt catctaagtt gtttttgagt 8820atattcgctt caatacaact cctctttata tttgatttaa gttttaaaat gcaacaacct 8880cgcggaagaa gtcgaacccg caaccaccaa gtcacaccga ctatatatca tgaaactcaa 8940ttgccctcca aacctcatta taccaattat catccacgtg caagatcgat gagctcaacc 9000cgtagtagtg cagaaagtag tcccaccaat catattcccc gtgctcgacc accctcaaca 9060ttcaacttat cgaaaccccc tcctcctcca aaagacatgt gcaggaacat gaaaattgga 9120ttgccgtgcg ctgatcccac ttgtaataga gatcatgacc ttgataatct aacaaatcgt 9180gaacttttgc tattgatggc ccgaaaaatg ctccccaata cagacaagac ctttagaagt 9240ccgcaggact gtggatcacc gtctctttct aaaggtctct caaaagataa acaggagcaa 9300acgaaagatg tgttgacctt ggaaaatcta ggacacattc tgagctatct ccacagatca 9360gaaattggga aattggatga gacatcactt cgtgcagcat taagtctgac gtgtgctgga 9420attcgaaaga cgaatagatc cttgatcaac accatgacag aattacacat gaaccatgaa 9480aatctcccgc aagaccaaaa cggtgttatc aagcagacct atacaggtat tcaccttgac 9540aaaggaggtc aattcgaagc cgccttatgg caaggttggg ataagagatc gatatctcta 9600ttcgtacaag cagctttata tgtaatgaac aatatcccct gtgaatcatc aatcagtgtg 9660caagcctcat acgatcattt tattcttcct caaagtcaag gtaaaggaca gtgattattg 9720ttcgaaagtt gacaatttga tcactttcag ttttcagttt caacccttat cgcgagactt 9780gaatacaatc ctactaactt caataagtga ccccaaattc aagtttgctg aaagctaaga 9840tgacaatgat cactagttca ttgtaaatta ctcgatcaaa atgttcttaa gctatcttaa 9900gcttactgat gcggctctgc ttcacttttc ttttgatttt aaagccatag ctatatctaa 9960gtgtctaatt aacaacttgt acctctaagg aaaaacatga agaacattaa gaaaaaggat 10020gttcttattc tttgactaaa cctgcatatt ctttgttgat accctcgaga gacaactttt 10080gacaccagat cacggatcaa gcacacttca atcaagcacc ctaaattttc aatcatacac 10140ataataacca ttttagtagc gttgcctttc agtacagtct aggtgattgt tgaaagactt 10200ccaagcatgg cagaattatc aacgcgttac aacttgcctg caaatgttac ggaaaatagt 10260ataaatcttg accttaattc cacagcacga tggataaaag aacccagtgt tgggggctgg 10320acagtgaagt ggggaaactt tgttttccat ataccaaata ctggaatgac attgttgcat 10380catttaaagt ctaacttcgt tgttccagag tggcaacaaa caaggaatct attctcccac 10440ctctttaaaa acccaaaatc aacaattata gaaccgtttt tggccctgag gattttgctt 10500ggagttgctt tgaaggatca agaattacag caatcattga ttcctggatt tagatctatt 10560gttcatatgc tatcagaatg gctgctcctg gaggtcacgt cggcaatcca tattagccct 10620aatctgttgg gaatctattt gacttcagac atgtttaaaa ttctgatggc aggtgtgaaa 10680aatttcttca ataagatgtt cactcttcat gttgtaaatg accacggaaa acccagcagt 10740attgaaataa agttaactgg acaacagatc attatcactc gtgttaatat ggggtttcta 10800gtggaagtca ggaggattga tattgaacct tgctgtggtg agacagtcct ctcagaatca 10860gttgtttttg gactagtggc tgaggcagtt ctaagagaac acagtcaaat ggagaagggc 10920caacctctca atctgacaca atacatgaac agcaaaattg ctatataagt ggcttaaatt 10980agcatgggta ttcctagttc gaccacataa taatgttgga ggcacagtac attatagtta 11040attgtcttgt atactaaggg atatacctaa cctgatttat atttactggt ataaaatagt 11100agcatcatct tattgaatag ttatcataca ataggctgtt cctataatct gattgtgaga 11160ttataaactt gtagaattac cgtgggtcac aactgttgca tatcctccaa aatatatctt 11220ttgcaagtga tgtgtgcttg aatacttcga tataatacat actaataacg attgattaag 11280aaaaatcaat gatggatatt aaatgtccat caagcaagtg ttgtagaata ccaggggttt 11340cacaggctgc taaacttact aaattttaca taggattata taattctttt cgatacacgt 11400tatatcttta gcaaagtgag gaaaacagct ttatcatgtt agatgccagt tatccatttt 11460aagtgaatcc tttcctcaac atgcagcatc caactcaata tcctgatgca aggttgtcct 11520ctcctataat cctagaccag tgtgacttat tagccagaag tttagggttg tatagtcatt 11580attcacataa tccgaaattg cgtaattgta ggattccaca tcacatttac cgtttaagga 11640attcgacagc attaaagaca tttcttcaga attgttcgat actcaccgtt ccttttcatt 11700caatttggga tcatatttta acttccattc aatatgatgc aattaatcat gttgatgatt 11760ttaaatacct attgccatct gagctagtca agtatgcgaa ttgggacaac gagttcttaa 11820aagcatatct taataagatc ttaggacttg accatgtttt ttcagcttct gcaaggtcac 11880agtgtgagga tttttctcct aaggaaaatc cttattattg ggggatgtta ttactcgtgc 11940atctatctca acttgccagg aggataaaag gacaaagagg gtcattaaga agtaactgga 12000aatttatagg aacagatttg gagctgtttg gaatagcaga ttttgttatt tttaaggttc 12060cagtaaaaac aataatccga aatgctgtaa gcttacaagc ttcaaaaccg ggattaagaa 12120tatggtaccg tgaccaaaac ttgacccctt atctatgcga cgatgagttt attgtaagcg 12180tcgctagtta tgaatgtttt atcatgatta aagacgtctt cattgagagg tataacacat 12240gggaaatctg tgcccgcgct tggctcgaag

acagtgatgg agctgattat cctcctcttg 12300atgtgttagg tgaattatac aaccagggag accaaattat tgccatgtac ctggaggacg 12360gtttcaaatt gattaaacac ttagaaccct tgtgtgtcag ctgtatacaa acacatggca 12420tctttacacc aagaaaatac tggttccaat cacagatgat taagtcatat tatgatgaac 12480tccatgatct caatttgaaa cttcaaattt cagacaataa ggctgagtgt gcccaaaact 12540ttattaaaac tatagttcag gcgaaattga ctcctcaaca atactgtgaa ttattctccc 12600tacaaaagca ttggggtcat cccgttttat acaatgatgt tgcactagat aaggttaaaa 12660aacatgcgca atcgacaaaa atcttaaaac ctaaagtcat gtttgaaact ttttgtgttt 12720tcaaatttat agtagcaaag aatcattatc attctcaagg atcatggtat aaaaccacac 12780atgatttgca tttgactcca tatcttagac aacatattgt gtcaaattca tttccatcac 12840aagccgaaat ttatcagcat ctttgggagt ggtatttcgt ggagcatgaa cctcttttct 12900caactaaaat aataagtgat ttaagtatct ttataaaaga cagggctacc gctgtgaacc 12960aggagtgttg ggacagtgtc ttcgatagaa gtgtattagg atataaccct cctgttagat 13020ttcaatcaaa gagagtgcca gagcaatttt tgggtcaagc agacttttcc ttgaatcaaa 13080tattagagtt tgctgaaaag ttagagtatt tggctccttc ttataggaat ttttccttct 13140cattaaaaga aaaagagttg aatataggaa gaacttttgg gaagttaccg tatcgtgtca 13200gaaatgtcca aacactcgca gaagccctgc tagcagatgg actggcaaaa gcattcccta 13260gtaacatgat ggttgtcact gagagggaac agaaagaagc attattacat caggcttctt 13320ggcaccacaa ttcagcaagc ataggggaga acgctatagt gaggggtgca agttttgtta 13380ctgatcttga gaaatacaac ctcgccttcc gatatgaatt tacacggcat ttcatagact 13440actgtaatcg atgttatggt gtgaagaatt tatttgattg gatgcatttt ttaataccac 13500tatgttatat gcatgtcagt gacttttata gcccaccaca ttgtgtgaca gaagataatc 13560gaaataaccc acctgattgt gctaatgctt atcattatca cttaggaggt atagagggac 13620ttcagcagaa attgtggaca tgcatatcat gtgcccaaat cacccttgtg gagttaaaaa 13680ctaaattaaa attgaagtcc agcgtcatgg gtgataatca atgtataaca actctaagtc 13740tttttccaat tgatgctccc aacgattatc aagagaacga ggctgaattg aatgcggcac 13800gagttgctgt cgaattagct attactacag gttatagtgg tatattttta aagcctgagg 13860aaacatttgt ccattcaggg ttcatttatt ttggtaaaaa gcaatatctc aacggtgttc 13920aactgccgca atcattgaaa acaatggcaa ggtgtggacc cttatctgac tctattttcg 13980atgatcttca aggttctctg gccagtattg gtacatcctt tgagagagga acaagcgaga 14040cacggcacat ttttccgagc cgttggatag cttcattcca ttcaatgtta gcaataaatt 14100tattaaatca gaatcacctt gggtttcctc tagggttcaa tattgatatt tcttgtttca 14160aaaagcctct taccttctcg gaaaaattaa ttgctctcat aacgccccaa gttttaggag 14220ggttatcatt tttaaatcca gaaaaattgt tctaccggaa cataagtgat cctctcactt 14280caggtctatt tcaactcaag aatgcattgg aatttcttga aaaggaagaa ttattctata 14340tcttgatttc taaaaaacct ggtttagcag atgcctcaga ttttgtcatg aatccattag 14400gcttaaatgt accaggatca aaggaaataa taacgttcct tagacaaaca gttcgcgaaa 14460atatcacgat cacgtcacaa aatagaataa taaattctct tttccacata ggttctgatt 14520tagaggacca aagagtgtgt gagtggcttt tatcatcaaa ccctgtaatg agccgatttg 14580ctgctgacat cttttcaaga acacctagtg gaaaacggct tcaagtctta ggctatctag 14640aaggaacaag aacattacta gcttctcgga caatcagttt aactacagaa ggaacaatgt 14700tgatgaaatt aagagaatta acgagaaacc gatggaaaag ttggttttct tatattgatg 14760cactggacga tgatttatct gagtccttgg aaaagttcac atgtactgtt gatgtggcta 14820atttcttgag ggcatattca tggtctgacg tcttaaaagg gaaaaggcta attggtgcca 14880cactgccatg tttactagag caatttgagg taaagtggat taatttatct gaggatttaa 14940gggaacaatt taatctatct tcagactcaa aatcaactat aaacttgttg ccgtatgact 15000gtaaggaact gcgacttgaa ggaagcaatg acacagagtt aaattatgtc agttgtgctc 15060ttgaccggaa agttgtccag aaacatccct ctgttaatcg tctagcttgg acgataggaa 15120atcgagcacc gtatattggc tcacggacag aagataagat cggttatcct cccttaagag 15180taaattgccc atcagcagca cttaaagaag ctattgagat ggtttctaga ttgttatggg 15240tgactcaagg cactgcagac cgagaaaaat tgcttattcc tcttctcaat tcaagagtaa 15300atctggacta tcagacggtg cttaactttt tacctacaca ctactcaggc aacatagttc 15360atagatataa tgatcaatat ggacaacatt cctttatggc aaacaggatg agtaatacat 15420ctacacgtgc aattatatca actaacacac tgggtaaata tgctggggga ggtcaagctg 15480ctattgatag taatataatc tttcaaaata ctattaattt aggagttgca gttttagata 15540ttgcattgtc tcttgctaaa ttgtcgtcag catcaaatgt cactttccgt ttaatgttaa 15600ataagtgctg cacgcggcat gtaccgtccg aatacctata ttttgataaa cctttagatg 15660tggatttgaa caagtatatg gacaatgagt tggtttatga caatgaccct ctttgcagtg 15720ggattaaagg gagattaggc agagtatctc gatcaacact cacattgagt ttgaatgtca 15780gtgacattgg ttcttatgac tttccaacta ttgctgcatg gacactagga gaaactatag 15840tcggaagcat tttttctgat gaatcttctc aaagtacgga tccaataagc tcaggttgca 15900caaaaacttt cgtcacacat ttccttgtgt atccagttga gagtatattt tatgcattcg 15960gagctaactt aatagttgaa agtttaagtc taagtaggat caaatcaatt aagaacctct 16020cagatttgac attccttata tcatccacaa tcaggaattt atcacataga tcacttcgga 16080ttcttcaatc tactttccga catgaattgg tgctcacccg actagcccac cacataccgt 16140taatttcttt aatgttaggg ggctctgcag gagagaagag ttcatcagat gctgttcggc 16200tatttcttac agcaagttat cagaatttta tcaataattt cagttgtctg atgaagaagg 16260gtcagtcatc gctaccggtt tggctttact ttcctagtga agggcaacaa ttaaagccta 16320tattaaaaat cttacagaga ttatcagact tgttatcacc tgacaaaatt caaaagcgta 16380aaattttggc tgacacctgt tgtccaattg gcagcttttg ggtctatcca agcaagtcca 16440caaggactaa ccattattat gcaagcctta attattggag agacaaagct aataaggtta 16500agaatactcc tttttcacac ttgataaatt gttcatttcc tgaattttct tcacatacca 16560gttcagtctc ctctaatcaa caagtgacca attcgaagta tattgtttat ccagaaaata 16620tcactgaaat aaatgcaaga accagattaa taaattatgg atcaacagct ctacagggga 16680tggacaccaa gatgccactc tcagagcaaa atctagtcga aaattgtcga ccatcagagg 16740gcatcagatt caaggacaat caaaaaataa caaaacatga ccagagatgt gagagggagg 16800aatcttcacc gcaacagatg ttccctgaag ataacatgca gactcctgcg cacatacata 16860gttcctcccc atttcaaatc cttataaaat cactagatgc acatgaggac tttgatgcct 16920cgaagataat cttaaattct gaaataaata atcttaacct tacggagtat actcttaata 16980caaagttatt gacaactcct accaggacag aaattttaga tacaagtccg ttacaatcct 17040ctagatattc atcaacttcc agggaacggt ctctactatc cagagaacaa gcttcatatt 17100tgtacgttga ttgcagtaat attccttcta tctctctaga cccaggtttt cggagtatgt 17160ctgatcagaa tcaagttcaa atgttaatca atacctacaa acgtgattta catgcttgtt 17220ttgatagcaa tcaattctgt cggtttacag gggtagtctc atcaatgcat tacaagcttt 17280atgatctttt gcctccaggt aaattgaaaa aggcaatttg tttggccgaa ggggaaggaa 17340gtggtgctcg gttacttttg aagtggaagg aaacggatta tttattcttc aacactttgg 17400ctacggattc acaacaagaa gccgagattt tgagtggccg ggtaataccg agaatgttgt 17460ataacataga cagattaagt gctttgcttg aatcaaggag actaatattg aacaacctaa 17520ctatccaaat tacagatatt acaaatccat tatggctaga ttctgtaata caatatttac 17580ctgaagatag tgacattctt acaatggacg cagagaccac caaggatgaa acaagggaac 17640agctttataa aactattgtg aatatttgga cacgtacttc tcctaatatc ccaaaaatta 17700gcatcatcaa ggtattttta ttagactatg aagggacttt attcttaatg aagaatgcta 17760ttcagtatta tgggcaagtt caactcaaga aaccatatag ctcaaatgca aaaaactcag 17820aatggtactt gtgttgcggt aaacgaagaa ttcaacggct ccaaattgat ttctcagacc 17880aggtgggaat ttttctgatt tgtaaagcaa tgtcacgcca aagacaagca attccttact 17940ggttaaaaca tatagaaaag aattatcctg cttcattaca cgagtttttc ctaactttgg 18000gtttcccttc tttagagtca tctttctgcc atcgttatac tattccattc agtgaaggaa 18060aggctctttt tcacaaggtc cagtcttatg ttcgtcaagg caaacaacat ttacattctc 18120ttatgttgga ttatgaaaac aattcacctc tactagactt gagaaatcac tttatttgct 18180cattaagggg aaagataact aaatattaca atgatatatt aaagttaaat ctagtcatca 18240aggcagtaga aaaaggtaaa aattggtcac aacttgttga gatccttcct aatatgcatt 18300cagtatgcat agtgcacgtg gatcatgagt gttctggatg cgagaaacgg ttattactta 18360aattggattt tatcagaaat acaaagatcg cagaacaaaa attacttaac agagtaatcg 18420ggtatatcct attctttcca ttcggtctgt ttaaatctgg atcattaagg gcataatttc 18480aacagagaga acttcattta attcacaaaa acaatctatt taagagtgag ggttacattg 18540tctaagatat tgtatgagaa gtaataaaat aaataagaaa acgaaaagac tattagacag 18600cttattttat acaagataat cttatatcgc tttaggcctc acacaagtga gaaaattacg 18660cgcacagatt aactagtgat tagtgtttgg tcacaccaga ggtaactttt taacgttaat 18720tactcagatg ttattgctca taattagcat taatattggc acattgggtg aatccttgag 18780ctttatccct aatatggtgt aagaaattaa ggaaatactg agatacacta gttgaattga 18840attatgacat accatatatc ataaatataa aaaagtgtct gctgtaatct acaagcacct 18900cttttaaata cattaggaaa agaattaagt taccgttgag atcaaaaaaa ccacgtcatg 18960ttttctctga tgacaagtga taaaacttcg tagttaaatt tctagaatgt cgatgtgaat 19020gtaaattaag aaaaaccaat atataaaatt aaaaaattaa aaagctttga tataagtaac 19080acaaaacatt cttcatcttt tttgtgtgtc c 1911116281PRTLake Victoria marburgvirus 16Met Gln Gln Pro Arg Gly Arg Ser Arg Thr Arg Asn His Gln Val Thr1 5 10 15 Pro Thr Ile Tyr His Glu Thr Gln Leu Pro Ser Lys Pro His Tyr Thr 20 25 30 Asn Tyr His Pro Arg Ala Arg Ser Met Ser Ser Thr Arg Ser Ser Ala 35 40 45 Glu Ser Ser Pro Thr Asn His Ile Pro Arg Ala Arg Pro Pro Ser Thr 50 55 60 Phe Asn Leu Ser Lys Pro Pro Pro Pro Pro Lys Asp Met Cys Arg Asn65 70 75 80 Met Lys Ile Gly Leu Pro Cys Ala Asp Pro Thr Cys Asn Arg Asp His 85 90 95 Asp Leu Asp Asn Leu Thr Asn Arg Glu Leu Leu Leu Leu Met Ala Arg 100 105 110 Lys Met Leu Pro Asn Thr Asp Lys Thr Phe Arg Ser Pro Gln Asp Cys 115 120 125 Gly Ser Pro Ser Leu Ser Lys Gly Leu Ser Lys Asp Lys Gln Glu Gln 130 135 140 Thr Lys Asp Val Leu Thr Leu Glu Asn Leu Gly His Ile Leu Ser Tyr145 150 155 160 Leu His Arg Ser Glu Ile Gly Lys Leu Asp Glu Thr Ser Leu Arg Ala 165 170 175 Ala Leu Ser Leu Thr Cys Ala Gly Ile Arg Lys Thr Asn Arg Ser Leu 180 185 190 Ile Asn Thr Met Thr Glu Leu His Met Asn His Glu Asn Leu Pro Gln 195 200 205 Asp Gln Asn Gly Val Ile Lys Gln Thr Tyr Thr Gly Ile His Leu Asp 210 215 220 Lys Gly Gly Gln Phe Glu Ala Ala Leu Trp Gln Gly Trp Asp Lys Arg225 230 235 240 Ser Ile Ser Leu Phe Val Gln Ala Ala Leu Tyr Val Met Asn Asn Ile 245 250 255 Pro Cys Glu Ser Ser Ile Ser Val Gln Ala Ser Tyr Asp His Phe Ile 260 265 270 Leu Pro Gln Ser Gln Gly Lys Gly Gln 275 280 1719114DNALake Victoria marburgvirus 17agacacacaa aaacaagaga tgatgatttt gtgtatcata taaataaaga agaatattaa 60cattgacatt aagactagtc attttgttaa tattctttaa aagatggatt tacatagttt 120gctagaatta ggtacaaaac ccacagcccc tcatgttcgt aataagaagg tgatattgtt 180tgatacaaac catcaggtta gtatctgtaa ccagataata gatgcaataa actcagggat 240cgatttagga gatcttttgg aaggcggctt gttgacatta tgtgttgaac attattacaa 300ttctgacaaa gataaattca atacaagtcc tattgcaaaa tatctgcggg acgcgggcta 360tgagtttgat gtcatcaaga atcctgatgc aactcgcttt ctagaggtta ttcccaatga 420acctcattac agccctttga ttttggctct caaaacctta gaaagcactg aatctcaaag 480ggggaggatt gggctctttc tgtcattttg cagtcttttt ctcccgaaac tcgttgtcgg 540agaccgagct agcatcgaaa aggccctgag acaagtaaca gtgcatcagg aacaagggat 600tgtaacatac cccaatcatt ggctcactac aggtcacatg aaagtaatct ttgggatttt 660aagatctagc ttcattttaa agtttgtctt aatccatcag ggagtaaact tggtgacagg 720tcatgatgcc tatgacagta tcatcagtaa ttcagtagga caaactagat tctcagggct 780tcttattgtg aaaacagttc tagagttcat cctacaaaaa actgattcag gggtggcatt 840gcatccactt gtgcggacct caaaagtaaa aaatgaagtt gcaagcttca aacaggcatt 900gagtaactta gctcgtcatg gagagtacgc accatttgca cgggttttga atttatcagg 960gatcaataac cttgagcatg ggctctatcc tcagctttca gcaattgcac tgggcgttgc 1020aacagcacat ggcagtacat tggctggtgt taacgttggc gaacaatacc aacaactgcg 1080agaagcagca catgatgcag aagtaaaatt acaaaggcga catgaacacc aggaaattca 1140ggccatcgcc gaggatgacg aagagagaaa aatattagaa cagttccatc tccagaagac 1200tgagattaca cacagtcaga cattggccgt cctcagccag aaacgagaga aactagctcg 1260tcttgctgca gaaattgaaa acaacattgc agaggatcag gggttcaaac aatcgcagaa 1320tcaggtgtca caatctttct taaatgatcc cacacctgta gaagtgacag tccaagccag 1380gtctataaat cgaccaacag ccctgccccc cccagtcgac aacaaaattg agcatgaaac 1440tgaagaggac agctcctcat caagtagttt tgttgacttg aatgatccat ttgcactgct 1500aaacgaagac gaggatactc ttgaaaatag tgtcatggcc ccaagcacta ctttgagaga 1560acccaaagaa gtttccgaac cactaaggca aactcaggac cttgatatta gccaaaagaa 1620acagggaaat gaatcaacag atccagcaag aaaacaattc ttgcgatatc aagaattacc 1680tcctgttcaa gaagacgatg aatcagaata cacaactgat tctcaggaaa gtgacgatca 1740accaggatct gataatgaac aaggcgttga tctcccacca cctccattat atgctcagga 1800aaagagacag gatcccatac agcatccaga cgtgagctcc caagatccct ttggcagtat 1860tggagatgta gatggtgata ttttggaacc tataagatcg ccttcttcac cgtctgctcc 1920tcaggaggac acaaggatgg gagaagccta tgaattatca cctgatttta caagctatga 1980ggataatcag cagaattggc cacagagagt ggtaacaaag aaaggcagga ccttccttta 2040tcctaatgac cttttacaga cgagtcctcc agagtcatta ataacagccc ttgttgaaga 2100gtaccaaaac cctgtctcag caaaagagct tcaagcggat tggcctgaca tgtcatttga 2160tgagaggagg catgttgcta tgaacttata atttcgacaa cacagcacga ctactcattt 2220atttacttca atatatttta cctccgaaac ataatagtca aatttattta aatatctaga 2280ccactttcaa ataccttgct atatatcact gttatgtgag atggtgcaaa ccttagaatg 2340ataaccaaag gtaggaactg gttatatgac agtacctcga aggtgttatt caatggttta 2400gatctctcct ctaattgcta cgagaataca actacaaacc tctttacctt ggttacaata 2460ctgtaataga tatgattgta tttctttctt aaatcatctg attcaacttg atagatataa 2520cttgattcag agaacatttt gggaacgtca ttaactaaat tctctaaatg atgtactgta 2580ttactgtttc acccgactaa ttatatagta gcatattaat ggatccttta cctctcctcc 2640tatgctcttc ttataagtca ctcaaccggt gaaacgccga gtttgttggc ttagagtttt 2700cctgttttac tgaatgtagt aattaatgat tgacttgagt gttgatggaa tcaagatagt 2760gcgatgattc atattataag gtacaatttc cattcctgtt ttgccaatgt atcttcttcc 2820ttatcacatg ccaattaaga aaaacaaaga gtcgaagaat attaaagatt ctctttaata 2880ttcaaaaaca gttcttaatt cttttccttt cctttattaa tataatatat cgataaatct 2940tacaatgtgg gactcgtcat acatgcaaca agtgagtgag ggactgatga ctggaaaagt 3000tccaatagat caagtgttcg gcactaatcc cttagaaaag ttatataaga gaagaaagcc 3060gaaagggaca gtgggattac aatgtagtcc ttgcttaata tcaaaatcaa caagtactga 3120cgacattgtt tgggatcagc taatcgtaaa gaaaacattg gctgacttgc ttatacctat 3180aaataggcaa atgtcggaca ttcaaagcac cctaagcgaa atgacaacaa aagtccatga 3240gatcgagcgt caactacatg atatcacccc agttgtaaaa atgggaaaaa cgctagaagc 3300aatttccaaa ggaatgtcag agatgctagc taagtacgat catctcgtga tttcaactgg 3360aagaaccacc gcaccagctg ctgcctttga tgcttactta aacgagcatg gagtcccccc 3420ccctcagcct gcaatcttca aagatcttgg agttgcccaa caagcctaca gtcaaaagac 3480tatggtcaaa aaccaaacaa cagatgcagc tgacaaaatg tcaaaggttc tggaactcag 3540tgaagaaaca ttttccaagc caaacctttc agctaaggat ttggctctat tattatttac 3600tcatctccct ggcaacaaca ctccattcca catactcgcc caagtccttt caaaaattgc 3660ttacaaatca ggaaagtctg gagcattctt ggatgcattc catcagattt taagcgaagg 3720ggagaatgct caggctgcat taactcgatt aagcagaaca ttcgatgctt tccttggagc 3780agttcctcca gtaataaaag ttaaaaactt tcaaacggtc ccccgccctt gtcaaaaaag 3840cctccgagct gttcccccaa atccaacaat tgacaaggga tgggtctgtg tctattcatc 3900tgaacagggt gaaacccggg ctcttaaaat ctaatccttg cgattcattc ttgaacaaag 3960aatgatcttt ctaggttaat aaaaaaccac taaacatttc aagagtttgt ggatgattta 4020agcatatttg ggtaaattta atcagggtag tagtttaaat ttgttttaag cgtgattttc 4080attaagagag ggttaattag ttatagattg atctttagtg tactccatat gcataatata 4140gagaaattaa tattactaac aaaaagggtt ttttaaacgg atttgattaa tagatcagta 4200tatctcaaaa gccaaataat tgacttctta ctctttggga atttactaac aatataggaa 4260agatatttat tcttacgtaa tccttggccc aacggaaact aacctccatg gtcattcaat 4320atgcttgctc atgatatatt cagagatttt ttataagttc aaaacgttgt aaattatact 4380tgcataaaat actgttttaa ttaagaaaaa ctatgaagaa cattaagtgg atttttcctt 4440cttagtgttc ttttacaaag caaggttttt aaattcaagt agatcaagtc tactcttgct 4500gaacttactt ctttaaaaat taatttacac taaacaattc gtttttgttg acggaacaaa 4560ttcagatatg gccagttcca gcaattataa tacgtatatg caatacctga accctccccc 4620ttatgctgat catggtgcaa atcagttaat cccagcagat cagctatcaa atcaacatgg 4680tataactccg aattatgtgg gcgatttgaa tctagatgat cagtttaaag ggaatgtttg 4740tcacgccttc actttggaag caataattga tatatctgct tataatgagc ggacggtcaa 4800aggagtccca gcgtggctgc ctcttgggat catgagcaat tttgaatacc ctttagccca 4860cactgttgct gcattgctta cagggagcta cacgattacc caattcacgc acaatgggca 4920aaaatttgtt cgtgttaatc gacttgggac aggaatccca gcacatccac tcagaatgct 4980gcgggaagga aaccaagctt ttgtccaaaa catggtgatt cccaggaact tttctacaaa 5040tcagtttacg tacaatctta ctaatctagt attgagcgtg caaaagcttc ctgatgatgc 5100ttggcgcccg tccaaagaca aattaatcgg aaacaccatg caccctgcgg tttctgtaca 5160cccaaacctg ccgcctattg tcctaccaac agttaaaaaa caagcctatc gtcagcataa 5220gaatcctaac aatggaccac tgctggccat atctggcatc cttcatcaac tgagggttga 5280aaaagtccca gagaagacga gcttattcag gatttcactt cctgccgaca tgttctcagt 5340aaaagagggc atgatgaaga aaagaggaga aggttctccg gtagtttatt tccaagcgcc 5400tgagaatttt cctttgaatg gcttcaacaa ccggcaagtt gtgctagcat atgctaaccc 5460gacactcagt gctgtttaat aagataattg ggtaagacaa tggcccttct gtacaaaggg 5520tctgattcag atagatattt gtcagattca tgcaggatca ttttaagttg attttaatag 5580tgctttaacc cttcactgct accctaaagg attgattgag ctgattaacc tataatgtat 5640aacttctttt aaaccgctaa atcaatcata agtttgtcag atcatatagg atgaatgtta 5700atacgtgata aatggttcct attcagtttt actttaacct catagtaaat cttataagac 5760tactcatctt caagttgatc aattcaaaga taatttccct tctaaaataa taagaaaaac 5820taatgaagaa cattaattgc taggtaaagg caattaagtt ctttgaactt tgcaaaagta 5880aggtttcact agtgagtaaa ttcctgtatt agcagattaa aaccaaggaa gcaccccgac 5940atgaagacca tatactttct gattagtctc attttaatcc aaagtataaa aactctccct 6000gttttagaaa ttgctagtaa cagccaacct caagatgtag attcagtgtg ctccggaacc 6060ctccaaaaga cagaagatgt tcatctgatg ggatttacac tgagtgggca aaaagttgct 6120gattcccctt tggaagcatc taaacgatgg gctttcagga caggtgttcc tcccaagaac 6180gttgagtata cggaaggaga agaagccaaa acatgttaca atataagtgt aacagaccct 6240tctggaaaat ccttgctgct ggatcctccc agtaatatcc gcgattaccc taaatgtaaa

6300actgttcatc atattcaagg tcaaaaccct catgcacagg ggattgccct ccatttgtgg 6360ggggcatttt tcttgtatga tcgcgttgcc tctacaacaa tgtaccgagg caaggtcttc 6420actgaaggaa atatagcagc tatgattgtt aataagacag ttcacagaat gattttttct 6480aggcaaggac aaggttatcg tcacatgaac ttgacctcca ccaataaata ttggacaagc 6540agcaatgaaa cgcggagaaa tgatacggga tgttttggca tcctccaaga atacaactcc 6600acaaacaatc aaacatgctc tccatctctt aaacctccat ccctgcccac agtaactccg 6660agcattcact ctacaaatac tcaaattaat actgctaaat ctggaactat gaacccaagt 6720agcgacgatg aggaccttat gatttccggc tcaggatctg gagaacaggg gccccacaca 6780actcttaatg tagtcactga acagaaacaa tcgtcaacaa tattgtccac tccttcacta 6840catccaagca cctcacaaca tgagcaaaac agtacgaatc cttcccgaca tgctgtaact 6900gagcacaatg gaaccgaccc aacaacacaa ccagcaacgc tcctcaacaa tactaatgca 6960actcccacct ataacactct caagtacaac ctcagtactc cttcccctcc aacccgcaac 7020atcaccaata atgatacaca acgtgaacta gcagaaagcg aacaagccaa tgctcagttg 7080aacacaactc tagatcctac agaaaatccc accacagcac aagacaccaa cagcacaacc 7140aacatcgtca tgacgacatc agatataaca agcaaacacc ccacaaattc ttctccggat 7200tctagtccga caacccgccc tcctatatac tttagaaaga aacgaagcat tttctggaaa 7260gaaggtgata tattcccgtt tttagatggg ttaataaata ctgaaattga ttttgatcca 7320atcccaaaca cagaaacaat ctttgatgaa tcccccagct ttaatacttc aactaatgag 7380gaacaacaca ctttcccgaa tatcagttta actttctctt attttcctga taaaaatgga 7440gatactgcct actctgggga aaacgaggat gattgtgatg cagagttgag gatttggagt 7500gtgcaggagg acgatttggc ggcagggctt agctggatac cattttttgg ccctggaatc 7560gaaggactct atactgccgg tttaatcaaa aatcagaaca atttagtttg taggttgagg 7620cgcttagcta atcaaactgc taaatccttg gagctcttgt taagggtcac aactgaggaa 7680aggacatttt ccttaatcaa taggcatgca attgactttt tgcttacgag gtggggcgga 7740acatgcaagg tgctaggacc tgattgttgc ataggaatag aagatctatc taaaaatatc 7800tcagaacaaa ttgacaaaat cagaaaggat gaacaaaagg aggaaactgg ctggggtcta 7860ggtggcaaat ggtggacatc tgactggggt gttctcacca atttgggcat cctgctacta 7920ttgtctatag ctgttctgat tgctctgtcc tgtatctgtc gtatcttcac taaatacatt 7980ggatgacata aagtttacaa tggttagagc tttaggaaag ttgctgctga gccctttgtc 8040taatctactg aaatcgactt aaagaatcct cagggagctt ataactcaat gtgaatcgat 8100tccctatata ttgttgattg tgctgatata gcagtcaagt gtcaccatca ttaggagcaa 8160ttcttctgat ccaatgcatt gaagtcacta attacttcta aaatccttat ctttaatccg 8220aatattcaca tagaaatcaa ttttgggaaa attccagtag tgcgtctttt catgtcactt 8280ctctgaacct aagaccataa taaatattgg ttaaagaggt cagctggtca atcttctatt 8340ttcatattaa agatatactg ctcagaaact cctagtttgc aacccaatct tggaaaaaga 8400aacctcctct tttgatcata actatctcta tcaatgtctt gaacacttaa tattattaac 8460aacttatttt tatttaatct tttaaatgca aatcaaaaga ggacatgagc caccctcact 8520ttacttcaat acgaaggaat tcttcttctc agcctacatt ttaatttacc aggtgataaa 8580cttatggtaa tttaccttaa cataggtgct gataagatga ttggatgata acacatttac 8640cgagatttaa tcgtatctca ttaagaaaaa gataattaga acactggaaa ttgataaact 8700tctattttga tcaatgtaga aaagaattat aaaaatctta gtaaataaat tactgcaaag 8760taaaaacgaa gaacattaag tgttctttat taaagttgtt catccttttt gcttttgatt 8820atatttgatc aaatacaact tcatttggta ttcattccaa gattcagaat gcaacagcct 8880cgtgggagaa gccgaaatcg tagccaccaa gttgcactat ccacatacca tgaaaatcaa 8940ttaccctcta aacctcaata cattaaccat catccacgtg caagatcaat gagttcaacc 9000cgtagtagta cagaaggtag ccctactaat catgcttccc gtgctcgacc actttcaaca 9060tttaatctat cgaaacctcc tccccccccg aaagacatgt gcaggaacat gaaaattggg 9120ttaccctgta ctgaccccgc ttgcaacagg gatcatgacc ttgataatct aacaaatcgt 9180gaactcttgc tgttgatggc acggaagatg ctccccaata cagataaggc tttcaaaagt 9240cagcaggact gtggatcgcc atctctttcc aaagggcttt caaaggacaa gcaggaacaa 9300gcaaaggatg tactgacttt ggaaaatcta gggcacatat tgaattatct tcatagatca 9360gaaatcggaa aattggacga gacatcactc cgtgcagcat tgagtttaac atgtgccgga 9420atccgaaaga caaataggtc tttgattaat actatgacag aattgcatat caaccatgag 9480aatcttccac aggaccaaaa tggtgttatt aagcagacat atacaggtat tcatcttgac 9540aaagggggtc aatttgaagc tgccttatgg cagggctggg acaagaagtc aatatctttg 9600tttgtgcaag ctgcattata tgtgatgaat aatatcccct gtgaatcgtc catcagtgtg 9660caggcctcat atgatcactt tattctccct cgaaatcagg gtgaaagaca atgattgtca 9720tttcaaaatc aacgatataa ttattgttaa cattctacct tggttcttat tgtaagactt 9780gtacactctc ctatcaactg tgattactag ttcaaaatta aaactcacaa aaattcaatc 9840gcattgtaat caattattta tcatcgatta tttagtttga gggattccac atcatcttaa 9900atccaataac acgattttgt ttgatttttc ttttaatttc tacaccataa caacatacaa 9960gtgtctgacg aacaacctgt gtttctatgg aaaaacatga agaacattaa gaaaaaggat 10020gttcttgtat ttcaaccaag gttgtatata tttggccgat atcctcggag aataattgtc 10080aatatctgac aattgatcat acacattcca ataatacatt atagactttt tatcagatat 10140agaactggga ttctaatagt tctacttctt agtgcagtac aggtttttgc aaagaaactt 10200ctgagtatgg cagaactgtc aacacgttac aacttaccaa caaatattac agagaagagc 10260ataaatcttg atctcaattc tactgcacgg tgggtaaaag agcccagtgt tgggggctgg 10320acagtgaaat gggggaattt tatctttcac atcccaaata ccggaatgac attgttgcat 10380catttaaaat ctaattttgt tgttccagaa tggcaacaaa caaggagtct cttctcccat 10440ctctttaaaa acccaaaatc aaccatcatg gagcctttct tggctttgag gatcttactt 10500ggcgttgctt tgaaagatca agaattgcag caatcattga ttcccggatt tagatccatt 10560gtgcatatgc tttcagaatg gctgctttta gaagttacgt cagctatcca tatcagtcct 10620aacctgttgg ggatctattt gacctcagac atgtttaaaa tcctaatggc aggtgtgaaa 10680aactttttta ataagctgtt taccctccat gttgtaaatg atcatggaaa acctagcagt 10740attgaaataa aattaaccgg acaacagatc ataatcactc gagttaacat ggggttctta 10800gtagaagtta ggaggattga cattgaacct tgctgcggcg agactgtcct ctcagaatca 10860gttgtgtttg ggctagtagc tgaagcagtc ctaagggaac acagccagat tgagagaggt 10920caacctctta atttgacaca atacatgaac agcaaaattg ctatttagac agcttgattc 10980ggcatctaaa gctttaattc agctgtataa ggatattgag aagacagtgg atcataatca 11040attgcccttt ataataaaag atgtagttgc cctaatttgt attcactaac ctaaaataat 11100agtattatca cactagatag tcatcacata ataagttgtt tctatgaatt aatcatgata 11160gtataatctt gtagaattac tatgggttct atatacggca tttattccgg actgtatcct 11220ttgtaaacaa tgcatgctta agtattctca tatcatacat gttatcactg cctgattaag 11280aaaaaccaat gatggatatt aaacatcctt caattgactg ttttgagact ttaaaatctt 11340acaggctgtc ttgatcttta aattctctac gggattatac agtcaccttt agcatacatt 11400atgcttctcg cgaaataata taagtgactc tatgacattc cgagccaggt ctttatcttg 11460aattaaccct ctttttggta tgcaacaccc gactcaatat ccagatgcaa gattatcttc 11520tcccataatt ttagatcagt gtgatttatt gaccagaagt ctagggttgt atagtcatta 11580ctcacacaat ccaaaactac gtaattgtag gattccatac cacatttatc gcttgaggaa 11640ttctacagca ttaaagacat ttcttcagaa ctgttcgata ctcacagttc cttttcactc 11700aatttgggat cacatcataa cttcaattca acacgatgca attaatcata tcaatgattt 11760caaataccta ttaccatcag aactcataaa gtatgctaat tgggacaatg agttcttaag 11820agtattcctt aacaagatct tgagactcga tcatgctttt acaaattctg caaagttaca 11880acgtgaggat ttctctccca aagagaatcc ttattattgg gggatgttat tgctcgtgca 11940tttatctcaa cttgccagaa ggattaaggg acaaagaggg tctttaagga gcaactggaa 12000gtttatagga gttgatttgg aactatttgg aatagcagat tttgttattt ttaaagttcc 12060gctaaaagca ataattcgga atgctacaag tttacaggcc tcaaaaccag ggttaaagac 12120atggtaccgt gatcaaaact taactcctta tctgtgtgat gatgaatttg ttgtaagcat 12180cgctagttat gaatgtttta tcatgattaa agatgtcttc atcgaaaggt acaacacatg 12240ggagatctgt gctcgcgctt gggtcgaaga taatgaagaa gctgattacc cacctcttgg 12300tatattaaga gatttgtaca atcaagggga ccaaattata accatgtatc tagaggatgg 12360tttcaaatta ataaaacact tagaaccttt atgtgtcagt tgtatacaaa cgtacggtat 12420ttttacgccg aggaagtact ggtttcaatc tcagatgatt aaatcatatt atgacgaact 12480tcgaagtctt aacctaaaac ttcagattcc ggataatagg actgaatgtg cacagaactt 12540tattaaaacc ataattcagg caaaactgac tcctcaacaa tactgtgaat tgttctcttt 12600acaaaaacat tggggtcacc cagttttata caatgatgtt gcactagaca aagtgaagaa 12660acatgcccaa tcaacaaaaa ttttaaaacc taaagtcatg tttgaaactt tttgtgtctt 12720taagtttata gtggcaaaaa atcattatca ctctcaagga tcgtggtaca aaaccacaca 12780tgatctacat ttgaccccat atctacggca gcacattgtg tcaaattcat ttccatcgca 12840agccgagatt tatcagcacc tctgggaatg gtactttgta gagcatgaac ctcttttttc 12900tacaaaaata ataagtgatt tgagtatttt cataaaagat agagctactg ctgtaaatcg 12960agagtgttgg gacagcgttt ttgataggag tgtgctagga tacaatcccc ctgtcagatt 13020tcaatcaaag agggtacctg agcaattctt aggtcaagca gatttctctt taaatcaaat 13080actggatttt gctgagaaat tagagtatct agctccttca tatagaaact tttccttttc 13140attaaaggaa aaagaattga atattgggag aacattcggg aagttgccct atcgtgtcag 13200aaatgtccaa acacttgcag aagccttatt agcagatgga ctagcgaagg cattccctag 13260taatatgatg gttgttactg agagagagca gaaagaagct ctattgcatc aagcctcttg 13320gcaccataat tcagcaagta taggggagaa tgccatagta agaggtgcaa gttttgttac 13380tgatcttgag aaatacaatc ttgcctttag atatgagttt acacgacatt tcatagacta 13440ctgtaataga tgttacggtg ttaaaaattt attcgactgg atgcatttct taataccact 13500atgctatatg catgtcagtg acttttatag ccctcctcac tgcgtgacag aaaataaccg 13560aaataaccca cctgattgtg ccaatgctta tcattatcat ttaggaggta tagaaggact 13620acaacagaaa ttgtggacat gtatatcatg tgctcaaatc acgcttgtag aactgaaaac 13680taagttaaaa ttgaaatcca gtgtcatggg tgataatcaa tgtataacaa ctctaagtct 13740tttccctatt gatgctccca atgattatca agaaaatgag gcagaattaa atgctgcacg 13800agttgctgtt gaattagcta ttactacagg ctatagtggg atattcttaa aacctgaaga 13860gacatttgtg cattcagggt ttatttactt tggtaaaaag caatacctaa atggtgttca 13920attaccacaa tcattgaaga caatggcaag gtgtggacct ttatcagatt ccattttcga 13980tgatcttcaa ggttcactag ccagtattgg cacatcattt gagagagggg caagcgagac 14040acggcatatt tttccaagtc gttggatagc tgcatttcat tccatgttag ccataaattt 14100gttaaatcag aatcacctcg gatttcccct aggatttagt attgatgtat cctgttttaa 14160aaagcctctt actttctcgg agaaattaat tgctttgatc acacctcaag tgctaggggg 14220attatctttc ttaaacccgg agaaattatt ctatcggaac atcagtgatc ctcttacttc 14280agggttattt caactcagga atgcattaga gttccttaga aaggaagagt tgttctacat 14340cttgattgct aaaaaacccg gtttagctga tgcttcagat ttcgttatga atccattagg 14400tttaaatgtg ccaggatcta gggaaataat aacgtttctc aggcaaacag ttcgtgaaaa 14460cataacgatt acatcacaaa atagaataat aaattctctt tttcacatag gttccgactt 14520agaggatcaa agagtatgtg aatggctttt atcatcaaac cccgtaatga gccgatttgc 14580tgctgatatt ttttcacgaa cacctagtgg aaaaagactt caagttttag gttacttgga 14640agggaccaga acgttattag cttcccgcac gatcagttta accactgagg gtacaatgtt 14700gatgagatta agagagttaa ctaagagtcg atggaagagt tggttttctt atattgatgc 14760attagatgat gatttgtctg agtctcttga aaagttcata tgcactgttg atgtggctaa 14820tttcttgaga gcatattcat ggtcagatgt cttgaaagga aagaagttaa ttggtgccac 14880actaccatgt ttactggagc aattcaatgt aaagtgggtc aacttgtctg aagacttaaa 14940ggagcaattt aagctatctt cagatctggg atcacctacg gatttattgc agtacgattg 15000caatggactg cattcaaagg gggccgataa cgcagaatta aattatgtga gttgtgccct 15060tgaccggaaa attgttcaaa agcatccatc tgacaatcgc ctggcatgga caataggaaa 15120tcgagcaccg tatatagggt cacgaacaga agataaaatt ggttaccctc ctttaagagt 15180aaactgccca tcggcagccc ttaaagaagc tattgaaatg gtctctagac tattgtgggt 15240gactcaaggc accgcagatc gagaaaaatt gctcattcct cttctcaatt caagagttaa 15300tttagactat cagacagtgc tcaacttcct gcccactcac tactcaggca atatagttca 15360cagatacaat gatcaatatg gacaacactc ctttatggca aatagaatga gcaatacatc 15420cactcgtgca atcatatcaa ctaacacact agggaaatat gctggagggg gtcaagctgc 15480tgttgatagt aatataatct tccagaacac tattaattta ggtgttgcag ttttggacat 15540tgcattgtct ctttctaaat tgtcatcaac atcaaatgtt tctttccgtt taatgttaag 15600taaatgttgc acacggcatg taccgtctga gtatttattc tttgataaac ctttagatgt 15660ggatttgaac aagtacatgg acaacgagtt agtttacgat aatgatcctc tctgtagtgg 15720aataaaggga agattaggta gagtatcaag atcaacactc tcattgagtc taaatgtaag 15780cgatattgga tcttacgact ttccgactat tgctgcgtgg actttaggag agacaattat 15840tggaagtatt ttttctgatg aatcttctca aagtacagac cctataagtt caggctgtac 15900aaaaactttt gtaacacact ttcttgtgta tccagttgag agtatttttt atgcctttgg 15960agctaatcta atagtggaaa gtttaagttt aagcaggatc aattcaatca agagcctctc 16020agatttaaca tttcttatat catccacaat cagaaatttg tcacacagat cacttcgaat 16080tcttcaatct actttccgac atgaattggt attaactaga ctagctcatc atataccatt 16140gatttcctta atgctaggag gttctgcggg tgagaaaagt tcatcggatg ctgtccgact 16200atttcttacg gcaagttatc aaaactttat caataatttc agttgtttga tgagaaagaa 16260ccaatcacca ttaccagttt ggctttattt ccctagtgaa gggcaacaac taaaacctat 16320tttaaaaatt ttgcaaaggt tatcatgttt attaacaact aaaaaggctc aaaatcacag 16380acctgtagct gatacttgtt ttttgactga taatttttgg gtctatccaa gcaaatcaac 16440gagaactaat cattattatg caagtcttaa ttattggaga gacaaagcta ataagattaa 16500gaatacttca ttttcacatt tgataaacta ttcattttct gaaccctctc tacatgcgag 16560ctctatctct tctagtcaag aagtggtcaa tttaaaacac actagtcgtt tagatgaaac 16620acctaatatg agtgaaaggg ctcaatcaac aaatcatgag ccaacagctt tacaagaggt 16680gtgcactgag atacccctct cggaacaaga tccagccaaa agttatttgc tgttagagaa 16740cactagattc agggataatc agaaaatatt aagacatgat cagaacgctg agaggggtga 16800acctctttca ttgcaagtgt cttctagggg ttgcctgcag gctcttactt gccctcatca 16860cccctcccca tctcaaacca ccacagaacc actaagcatg cttaggaatt gtgacgccat 16920aaaagcagcc ttacgttctg agacgaatga tccccgtctt atgagcagta tccttgatat 16980gagatcattg aaaactccca tgagaataga atctcgaaac acgagtctat tgcaaccttc 17040tgagtgtctg tcaacttcta agggaaaatc tgtactgtct agagaacagg cttcatacct 17100gtatgttgat tgcagtaata tctcttctat ttctctggat tcaggttttc gaaatatgtc 17160tgatagaaat caagtccaaa tgctaataaa tacttacaaa cgcgacttat acacttgttt 17220tgatagtaac caattctgca ggtttacagg ggtcgtttca tcaatgcatt ataagcttta 17280tgatcttttg ccagcaggca aactcggaaa ggcaatctgc ctagccgaag gggaagggag 17340tggcgctcga ctactcttga agtggaagga gacagattat ttattcttca atactttggc 17400cacagattca caacaggaag cagaaatttt gagtggtcga gttattccaa ggatgttgta 17460taacatagat aagctaagtg ttttacttga atccagaaaa ttaatcttga ataatctaac 17520tattcaaatc acggatatta cacacccact atggctggac tctgtcatac aatacctacc 17580tgaagatagt gacattctaa caatggatgc agagaccact aaagaagaga caagagagca 17640actctataaa actatcataa atatttgggc acgtacttct cctaatatcc ctaaaaccag 17700catcattaaa gtgtttttat tagattatgg gggaaccttg ttcttaatga agaatgctat 17760tcaatattat ggacaagttc aacttaagaa accatatagt tcaaatgcaa aaaattcaga 17820atggtactta tgttgtggaa aacgaagagt tcaacgactc cgagttgatt ttccagacca 17880agtaggaata ttcttgatct gtaaagcaat gtcacgtcag aggcaagcaa ttccttactg 17940gctaaagcac atagaaaaga attaccctgc ttcattgcac gagttcttta taactttagg 18000ttttccttct ttagagtcat ctttctgcca tcgctacacc attccgttca ctgagggaac 18060ggctctcttt cacaaggtcc agtcttacgt ccgacaaggt agacaacacc tacactctct 18120tatgttagat tacgaaaata attcacccct cctagatctg agaaatcact tcatatgctc 18180attgagggga aagatagcca agtattacaa tgacatattg aaattaagtt tagtagtgag 18240agcagtagaa agagggaaaa attggtcgca actcgttgag tcccttccta atatgcactc 18300agtatgcata acacatgttg atcacgaatg tattggctgt gagagacggt tattacttaa 18360attggacttt gtcagaaata caaagatagc agaacaaaag ttactcaata gggtaattgg 18420gtatattcta ttcttccctt ttggtttctc cagacccaag tgactacaga tatattcttc 18480aataaaggaa gctcagtcta actcacagaa ataatccact tcaaaacaag gatcacccat 18540tttggaacat tgtataagaa actgcaagac aaataataag gaaaggatac tactgtataa 18600cttgttatat ccaaaaggat cttgggtcat tttaagcatg atgcaaataa aaaatcgtct 18660acatagccga actgaccgct cagtacttat tcacaatcat gctaactttt agtttttaat 18720tgtgcaaaaa ttactaagaa taattaatat tgatattaaa acattaaatg gacatttgag 18780ttttatgcct agaataatat aaagaaattt aagagatatt tagatatatc agttgaattg 18840atttatgaca catagtgcat catgaataca aagagaaaaa tcgttgcaat tcaggaatat 18900cttatttaaa tgtattagag agaaagtcag attattatca aaatcaagca aaatacaata 18960ggttttttca aagaataggt ggtaaagcct tatggttatt ttttaaagat gtcaatgtga 19020atttttatta agaaaaagta atgcatgaaa ttaaaaaatt aaagaacttt gatataagta 19080acacaaaaca ctcttcatct ttttagtgtg tcca 1911418281PRTLake Victoria marburgvirus 18Met Gln Gln Pro Arg Gly Arg Ser Arg Asn Arg Ser His Gln Val Ala1 5 10 15 Leu Ser Thr Tyr His Glu Asn Gln Leu Pro Ser Lys Pro Gln Tyr Ile 20 25 30 Asn His His Pro Arg Ala Arg Ser Met Ser Ser Thr Arg Ser Ser Thr 35 40 45 Glu Gly Ser Pro Thr Asn His Ala Ser Arg Ala Arg Pro Leu Ser Thr 50 55 60 Phe Asn Leu Ser Lys Pro Pro Pro Pro Pro Lys Asp Met Cys Arg Asn65 70 75 80 Met Lys Ile Gly Leu Pro Cys Thr Asp Pro Ala Cys Asn Arg Asp His 85 90 95 Asp Leu Asp Asn Leu Thr Asn Arg Glu Leu Leu Leu Leu Met Ala Arg 100 105 110 Lys Met Leu Pro Asn Thr Asp Lys Ala Phe Lys Ser Gln Gln Asp Cys 115 120 125 Gly Ser Pro Ser Leu Ser Lys Gly Leu Ser Lys Asp Lys Gln Glu Gln 130 135 140 Ala Lys Asp Val Leu Thr Leu Glu Asn Leu Gly His Ile Leu Asn Tyr145 150 155 160 Leu His Arg Ser Glu Ile Gly Lys Leu Asp Glu Thr Ser Leu Arg Ala 165 170 175 Ala Leu Ser Leu Thr Cys Ala Gly Ile Arg Lys Thr Asn Arg Ser Leu 180 185 190 Ile Asn Thr Met Thr Glu Leu His Ile Asn His Glu Asn Leu Pro Gln 195 200 205 Asp Gln Asn Gly Val Ile Lys Gln Thr Tyr Thr Gly Ile His Leu Asp 210 215 220 Lys Gly Gly Gln Phe Glu Ala Ala Leu Trp Gln Gly Trp Asp Lys Lys225 230 235 240 Ser Ile Ser Leu Phe Val Gln Ala Ala Leu Tyr Val Met Asn Asn Ile 245 250 255 Pro Cys Glu Ser Ser Ile Ser Val Gln Ala Ser Tyr Asp His Phe Ile 260 265 270 Leu Pro Arg Asn Gln Gly Glu Arg Gln 275 280 1919114DNALake Victoria marburgvirus 19agacacacaa aaacaagaga tgatgatttt gtgtatcata taaataaaga agaatattaa 60cattgacatt aagactagtc attttgttaa tattctttaa aagatggatt tacatagttt 120gctagaatta ggtacaaaac ccacagcccc tcatgttcgt aataagaagg tgatattgtt 180tgatacaaac catcaggtta gtatctgtaa ccagataata gatgcaataa actcagggat 240cgatttagga gatcttttgg aaggcggctt gttgacatta tgtgttgaac attattacaa 300ttctgacaaa gataaattca

atacaagtcc tattgcaaaa tatctgcggg acgcgggcta 360tgagtttgat gtcatcaaga atcctgatgc aactcgcttt ctagaggtta ttcccaatga 420acctcattac agccctttga ttttggctct caaaacctta gaaagcactg aatctcaaag 480ggggaggatt gggctctttc tgtcattttg cagtcttttt ctcccgaaac tcgttgtcgg 540agaccgagct agcatcgaaa aggccctgag acaagtaaca gtgcatcagg aacaagggat 600tgtaacatac cccaatcatt ggctcactac aggtcacatg aaagtaatct ttgggatttt 660aagatctagc ttcattttaa agtttgtctt aatccatcag ggagtaaact tggtgacagg 720tcatgatgcc tatgacagta tcatcagtaa ttcagtagga caaactagat tctcagggct 780tcttattgtg aaaacagttc tagagttcat cctacaaaaa actgattcag gggtggcatt 840gcatccactt gtgcggacct caaaagtaaa aaatgaagtt gcaagcttca aacaggcatt 900gagtaactta gctcgtcatg gagagtacgc accatttgca cgggttttga atttatcagg 960gatcaataac cttgagcatg ggctctatcc tcagctttca gcaattgcac tgggcgttgc 1020aacagcacat ggcagtacat tggctggtgt taacgttggc gaacaatacc aacaactgcg 1080agaagcagca catgatgcag aagtaaaatt acaaaggcga catgaacacc aggaaattca 1140ggccatcgcc gaggatgacg aagagagaaa aatattagaa cagttccatc tccagaagac 1200tgagattaca cacagtcaga cattggccgt cctcagccag aaacgagaga aactagcccg 1260tcttgctgca gaaattgaaa acaacattgc agaggatcag gggttcaaac aatcgcagaa 1320tcaggtgtca cagtctttct taaatgatcc cacacctgta gaagtgacag tccaagccag 1380gtctataaat cgaccaacag ccctgccccc cccagtcgac aacaaaattg agcatgaaac 1440tgaagaggac agctcctcat caagtagttt tgttgacttg aatgatccat ttgcactgct 1500gaacgaagac gaggatactc ttgaaaatag tgtcatggcc ccaagcacta ctttgagaga 1560acccaaagaa gtttccgaac cactaaggca aactcaggac cttgatatta gccaaaagaa 1620acagggaaat gaatcaacag atccagcaag aaaacaattc ttacgatatc aagaattacc 1680tcctgttcaa gaagacgatg aatcagaata cacaactgat tctcaggaaa gtgacgatca 1740accaggatct gataatgaac aaggtgttga tctcccacca cctccattat atgctcagga 1800aaagagacag gatcccatac agcatccagc cgtgagctcc caagatccct ttggcagtat 1860tggtgatgta gatggtgata ttttggaacc tataagatcg ccttcctcac cgtctgctcc 1920tcaggaggac acaaggatgg gagaagccta tgaattatca cctgatttta caagctatga 1980ggataatcag cagaattggc cacagagagt ggtaacaaag aaaggcagga ccttccttta 2040tcctaatgac cttttacaga cgagtcctcc agagtcatta ataacagccc ttgttgaaga 2100gtaccaaaac cctgtctcag caaaagagct tcaagcggat tggcctgaca tgtcatttga 2160tgagaggagg catgttgcta tgaacttgta atttcgacaa cacagcacga ctactcattt 2220atttacttca atatatttta cctccgaaac ataatagtca aatttattta aatatctaga 2280ccactttcaa ataccttgtt atatatcact gttatgtgag atggtgcaaa ccttagaatg 2340ataaccaaag gtaggaactg gttatatgac agtacctcga aggtgttatt caatggttta 2400gatctctcct ctaattgcta cgataataca actacaaacc tctttacctt ggttacaata 2460ctgtaataga tatgattgta tttctttctt aaatcatctg attcaacttg atagatataa 2520cttgattcag agaacatttt gggaacgtca ttaactaaat tctctaaatg atgtactgta 2580ttactgtttc acccgactaa ttatatagta gcatattaat ggatccttta cctctcctcc 2640tatgctcttc ttataagtca ctcaaccggt gaaacgccga gtttgttggc ttagagtttt 2700cctgttttac tgaatgtagt aattaatgat tgacttgagt gttgatggaa tcaagatagt 2760gcgatgattc atattataag gtacaatttc catttctgtt ttgccaatgt atcctcttcc 2820ttatcacatg ccaatcaaga aaaacaaaga gtcgaagaat attaaagatt ctctttaata 2880ttcaaaaaca gttcttaatt cttttccttt cctttattaa tataatatat cgataaatct 2940tacaatgtgg gactcgtcat acatgcaaca agtgagtgag ggactgatga ctggaaaagt 3000tccaatagat caagtgttcg gcactaatcc cttagaaaag ttatataaga gaagaaagcc 3060gaaagggaca gtgggattac aatgtagtcc ttgcttaata tcaaaatcaa caagtactga 3120cgacattgtt tgggatcagc taatcgtaaa gaaaacattg gctgacttgc ttatacctat 3180aaataggcaa atgtcggaca ttcaaagcac cctaagcgaa atgacaacaa aagtccatga 3240gatcgagcgt caactacatg atatcacccc agttgtaaaa atgggaaaaa cgctagaagc 3300aatttccaaa ggaatgtcag agatgctagc taagtacgat catctcgtga tttcaactgg 3360aagaaccacc gcaccagctg ctgcctttga tgcttactta aacgagcatg gagtcccccc 3420ccctcagcct gcaatcttca aagatcttgg agttgcccaa caagcctaca gtcaaaagac 3480tatggtcaaa aaccaaacaa cagatgcagc tgacaaaatg tcaaaggttc tggaactcag 3540tgaagaaaca ttttccaagc caaacctttc agctaaggat ttagctctat tattatttac 3600tcatctccct ggcaacaaca ctccattcca catactcgcc caagtccttt caaaaattgc 3660ttacaaatca ggaaagtctg gagcattctt ggatgcattc catcagattt taagcgaagg 3720ggagaatgct caggctgcat taacccgatt aagcagaaca ttcgatgctt tccttggagc 3780agttcctcca gtaataaaag ttaaaaactt tcaaacggtc ccccgccctt gtcaaaaaag 3840cctccgagct gttcccccaa atccaacaat tgacaaggga tgggtctgtg tctattcatc 3900tgaacagggt gaaacccggg ctcttaaaat ctaatccttg cgattcattc ttgaacaaag 3960aatgatcttt ctaggttaat acaaaaacac taaacatttc aagagtttgt ggatgattta 4020agcatatttg ggtaaattta atcaggatag tagtttaaat ttgttttaag cgtgattttc 4080attaagagag ggttaattag ttatagattg atctttagtg tactccatac gcataatata 4140gagaaattaa tattactaac aaaaagggtt ttttaaacgg atttgattaa tagatcagta 4200tatctcaaaa gccaaataat tgacttctta ctctttggga atttactaac aatataggaa 4260agatatttat tcttacgtaa tccttggccc aacggaaact aacctcaatg gtcattcaat 4320atgcttgctc atgatatatc cagagatttt ttataagttc aaaacgttgt aaattatact 4380tgcataaaat actgttttaa ttaagaaaaa ctatgaagaa cattaagtgg atttttcctt 4440cttagtgttc ttttacaaag caaggttttt aaattcaagt agatcaagtc tactcttgct 4500gaacttactt ctttaaaaat taatttacac taaacaattc gtttttgttg acggaacaaa 4560ttcagatatg gccagttcca gcaattataa tacgtatatg caatacctga accctccccc 4620ttatgctgat catggtgcaa atcagttaat cccagcagac cagctatcaa atcaacatgg 4680tataactccg aattatgtgg gcgacttgaa tctagatgat cagtttaaag ggaatgtttg 4740tcacgccttc actttggaag caataattga tatatctgct tataatgagc ggacggtcaa 4800aggagtccca gcgtggctgc ctcttgggat catgagcaat tttgaatacc ctttagccca 4860cactgttgct gcattgctta cagggagcta cacgattacc caattcacgc acaatggaca 4920aaaatttgtt cgtgttaatc gacttgggac aggaatccca gcacatccac tcagaatgct 4980gcgggaagga aaccaagctt ttgtccaaaa catggtgatt cccaggaact tttctacaaa 5040tcagtttacg tacaatctta ctaatctagt attgagcgtg caaaagcttc ctgatgatgc 5100ttggcgcccg tccaaagaca aattaatcgg aaacaccatg caccctgcgg tttctgtaca 5160cccaaacctg ccgcctattg tcctaccaac agttaaaaaa caagcctatc gtcagcataa 5220gaatcctaac aatggaccac tgctggccat atctggcatc cttcatcaac tgagggttga 5280aaaagtccca gagaagacga gcttattcag gatttcactt cctgccgaca tgttctcagt 5340aaaagagggc atgatgaaga aaagaggaga aggttctccg gtagtttatt tccaagcgcc 5400tgagaatttt cctttgaatg gcttcaacaa ccggcaagtt gtgctagcat atgctaaccc 5460gacactcagt gctgtttaat aagataattg ggtaagacaa tggcccttct gtacaaaggg 5520tctgattcag atagatattt gtcagattca tgcaggatca ttttaagttg attttaatag 5580tgctttaacc cttcactgct accctaaagg attgattgag ctgattaacc tataatgtat 5640aacttctttt aaaccgctaa atcaatcata agtttgtcag atcatatagg atgaatgtta 5700atacgtgata aatggttcct attcagtttt actttaacct catagtaaat cttataagac 5760tactcatctt caagttgatc aattcaaaga taatttccct tctaaaataa taagaaaaac 5820taatgaagaa cattaattgc taggtaaagg caattaagtt ctttgaactt tgcaaaagta 5880aggtttcact agtgagtaaa ttcctgtatt agtagattaa aaccaaggaa gcaccccgac 5940atgaagacca tatattttct gattagtctc attttaatcc aaagtataaa aactctccct 6000gttttagaaa ttgctagtaa cagccaacct caagatgtag attcagtgtg ctccggaacc 6060ctccaaaaga cagaagatgt tcatctgatg ggatttacac tgagtgggca aaaagttgct 6120gattcccctt tggaagcatc taaacgatgg gctttcagga caggtgttcc tcccaagaac 6180gttgagtata cggaaggaga agaagccaaa acatgttaca atataagtgt aacagaccct 6240tctggaaaat ccttgctgct ggatcctccc agtaatatcc gcgattaccc taaatgtaaa 6300actgttcatc atattcaagg tcaaaaccct catgcacagg ggattgccct ccatttgtgg 6360ggggcatttt tcttgtatga tcgcgttgcc tctacaacaa tgtaccgagg caaggtcttc 6420actgaaggaa atatagcagc tatgattgtt aataagacag ttcacagaat gattttttct 6480aggcaaggac aaggttatcg tcacatgaac ttgacctcca ccaataaata ttggacaagc 6540agcaatgaaa cgcagagaaa tgatacggga tgttttggca tcctccaaga atacaactcc 6600acaaacaatc aaacatgccc tccatctctt aaacctccat ccctgcccac agtaactccg 6660agcattcact ctacaaatac tcaaattaat actgctaaat ctggaactat gaacccaagt 6720agcgacgatg aggaccttat gatttccggc tcaggatctg gagaacaggg gccccacaca 6780actcttaatg tagtcactga acagaaacaa tcgtcaacaa tattgtccac tccttcacta 6840catccaagca cctcacaaca tgagcaaaac agtacgaatc cttcccgaca tgctgtaact 6900gagcacaatg gaaccgaccc aacaacacaa ccagcaacgc tcctcaacaa tactaataca 6960actcccacct ataacactct caagtacaac ctcagtactc cttcccctcc aacccgcaac 7020atcaccaata atgatacaca acgtgaacta gcagaaagcg aacaaaccaa tgctcagttg 7080aacacaactc tagatccaac agaaaatccc accacaggac aagacaccaa cagcacaacc 7140aacatcatca tgacgacatc agatataaca agcaaacacc ccacaaattc ttctccggat 7200tctagtccga caacccgccc tcctatatac tttagaaaga aacgaagcat tttctggaaa 7260gaaggtgata tattcccgtt tttagatggg ttaataaata ctgaaattga ttttgatcca 7320atcccaaaca cagaaacaat ctttgatgaa tctcccagct ttaatacttc aactaatgag 7380gaacaacaca ctcccccgaa tatcagttta actttctctt attttcctga taaaaatgga 7440gatactgcct actctgggga aaacgagaat gattgtgatg cagagttgag gatttggagt 7500gtgcaggagg acgatttggc ggcagggctt agctggatac cattttttgg ccctggaatc 7560gaaggactct atactgccgg tttaatcaaa aatcagaaca atttagtttg taggttgagg 7620cgcttagcta atcaaactgc taaatccttg gagctcttgt taagggtcac aaccgaggaa 7680aggacatttt ccttaatcaa taggcatgca attgactttt tgcttacgag gtggggcgga 7740acatgcaagg tgctaggacc tgattgttgc ataggaatag aagatctatc taaaaatatc 7800tcagaacaaa tcgacaaaat cagaaaggat gaacaaaagg aggaaactgg ctggggtcta 7860ggtggcaaat ggtggacatc tgactggggt gttctcacca atttgggcat cctgctacta 7920ttatctatag ctgttctgat tgctctgtcc tgtatctgtc gtatcttcac taaatacatt 7980ggatgacata aagtttacaa tggttagagc tttaggaaag ttgctgctga gccctttgtc 8040taatctactg aaatcgactt aaagaatcct cagggagctt ataactcaat gtgaatcgat 8100tccctatata ttgttgatcg tgatgatata gcagtcaagt gtcaccatca ttaggagcaa 8160ttcttctgat ccaatgcatt gaagtcacta attacttcta aaatccttat ctttaatccg 8220aatattcaca tagaaatcaa ttttgggaaa attccagtag tgcgtctttt catgtcactt 8280ctctgaacct aagaccataa taaatattgg ttaaagaggt cagctggtca atcttctatt 8340ttcatattaa agatatactg ctcagaaact cctagtttgc aacccaatct tggaaaaata 8400aacctcctct tttgatcata actatctcta tcaatgtctt gaacacttaa tattattaac 8460aacttatttt tatttaatct tttaaatgca aatcaaaaga ggacatgagc caccctcatt 8520ttacttcaat acgaaggaat tcttcttctc agcctacatt ttaatttacc aggtgataaa 8580cttttggtaa tttaccttaa cataggtgct gataagatga ttggatgata acacatttac 8640cgagatttaa tcgtatctca ttaagaaaaa gataattaga acactggaaa ttgataaact 8700tctattttga tcaatgtaga aaagaattat aaaaatctta gtaaataaat tactgcaaag 8760taaaaacgaa gaacattaag tgttctttat taaaattgtt catccttttc gcttttgatt 8820atatttgatc aaatacaact tcatttggta ttcattccaa gattcagaat gcaacagcct 8880cgtgggagaa gccgaaatcg tagccaccaa gttgcactat ccacatacca tgaaaatcaa 8940ttaccctcta aacctcagta cattaaccat catccacgtg caagatcaat gagttcaacc 9000cgtagtagta cagaaggtag ccctactaat catgcttccc gtgctcgacc actttcaaca 9060tttaatctat cgaaacctcc tccccccccg aaagacatgt gcaggaacat gaaaattggg 9120ttaccctgta ctgaccccgc ttgcaacagg gatcatgacc ttgataatct aacaaatcgt 9180gaacttttgc tgttgatggc acggaagatg ctccccaata cagataaggc tttcaaaagt 9240cagcaggact gtggatcgcc atctctttcc aaagggcttt caaaggacaa gcaggaacaa 9300gcaaaggatg tactgacttt ggaaaatcta gggcacatat tgaattatct tcatagatca 9360gaaatcggaa aattggacga gacatcactc cgtgcagcat tgagtttaac atgtgccgga 9420atccgaaaga caaataggtc tttgattaat actatgacag aattgcatat caaccatgag 9480aatcttccac aggaccaaaa tggtgttatt aagcagacat atacaggtat tcatcttgac 9540aaagggggtc aatttgaagc tgccttatgg cagggctggg acaagaagtc aatatcgttg 9600tttgtgcaag ctgcattata tgtgatgaat aatatcccct gtgaatcgtc catcagtgtg 9660caggcctcat atgatcactt tattctccct cgaaatcagg gtgaaagaca atgattgtca 9720tttcaaaatc aacgatataa ttattgttaa cattctacct tggttcttat tgtaagactt 9780gtatactctc ctatcaactg tgactactag ttcaaaatta aaactcacaa aaattcaatc 9840gcattgtaat caattattta tcatcgattg tttagtttga gggattccac atcatcttaa 9900atccaataac acgattttgt ttgatttttc ttttaatttc tacaccacaa caacatacaa 9960gtgtctgacg aacgacctgt gtttctatgg aaaaacatga agaacattaa gaaaaaggat 10020gttcttgtat ttcaaccaag gttgtatata tttggccgat atcctcggag aataattgtc 10080aatatctgac aattgatcat acacattcca ataatacatt atagactttt tatcatatat 10140agaactggga ttctaatagt tctacttctt agtgcagtac aggtttttgc aaagaaactt 10200ctgagtatgg cagaactgtc aacacgttac aacttaccaa caaatattac agagaagagc 10260ataaatcttg atctcaattc tactgcacgg tgggtaaaag agcccagtgt tgggggctgg 10320acagtgaaat gggggaattt tatctttcac atcccaaata ccggaatgac attgttgcat 10380catttaaaat ctaattttgt tgttccagaa tggcaacaaa caaggagtct cttctcccat 10440ctctttaaaa acccaaaatc aaccatcatg gagcctttct tggctttgag gatcttactt 10500ggcgttgctt tgaaggatca agaattgcag caatcattga ttcctggatt tagatccatt 10560gtgcatatgc tttcagaatg gctgctttta gaagttacgt cagctatcca tatcagtcct 10620aacctgttgg ggatctattt gacctcagac atgtttaaaa tcctaatggc aggtgtgaaa 10680aactttttca ataagctgtt taccctccat gttgtaaatg atcatggaaa acctagcagt 10740attgaaataa aattaaccgg acaacagatc ataatcactc gagttaacat ggggttctta 10800gtagaagtta ggaggattga cattgaacct tgctgcggcg agactgtcct ctcagaatca 10860gttgtgtttg ggctagtagc tgaagcagtc ctaagggaac acagccagat tgagagaggt 10920caacctctta atttgacaca atacatgaac agcaaaattg ctatttagac agcttgattc 10980ggcatctaaa gctttaattc agctgtataa ggatattgag aagacagtgg atcataatca 11040attgcccttt ataataaaag atgtagttgc cctaatttgt attcactaac ctaaaataat 11100agtattatca cactagatag tcatcacata ataagttgtt tctatgaatt aatcatgata 11160gtataatctt gtagaattac tatgggttct atatacggca tttattccgg actgtatcct 11220ttgtaaacaa tgcatgctta agtattctca tatcatacat gttatcactg cctgattaag 11280aaaaaccaat gatggatatt aaacatcctt caattgactg ttttgagact ttaaaatctt 11340acaggctgtc ttgattttta aattctctac gggattatac agtcaccttt agcatacatt 11400atgcctctcg cgaaataata taagtgactc tatgacattc cgagccaggt ctttatcttg 11460aattaaccct ctttttggta tgcaacaccc gactcaatat ccagatgcaa gattatcttc 11520tcccataatt ttagatcagt gtgatttatt gaccagaagt ctagggttgt atagtcatta 11580ctcacacaat ccaaaactac gtaattgtag gattccatac cacatttatc gcttgaggaa 11640ttctacagca ttaaagacat ttcttcagaa ctgttcgata ctcacagttc cttttcactc 11700aatttgggat cacatcataa cttcaattca acacgatgca attaatcata tcaatgattt 11760caaataccta ttaccatcag aactcataaa gtatgctaat tgggacaatg agttcttaag 11820agtattcctt aacaagatct tgagactcga tcatgctttt acaaattctg caaagttaca 11880atgtgaggat ttctctccca aagagaatcc ttattattgg gggatgttat tgctcgtgca 11940tttatctcaa cttgccagaa ggattaaggg acaaagaggg tctttaagga gcaactggaa 12000gtttatagga gttgatttgg aactatttgg aatagcagat tttgttattt ttaaagttcc 12060gataaaagca ataattcgga atgctacaag tttacaggcc tcaaaaccag ggttaaagac 12120atggtaccgt gatcaaaact taactcctta tctgtgtgat gatgaatttg ttgtaagcat 12180cgctagttat gaatgtttta tcatgattaa agatgtcttc atcgaaaggt acaacacgtg 12240ggagatctgt gctcgcgctt gggtcgaaga taatgaagaa gctgattacc cacctcttgg 12300tatattaaga gatttgtaca atcaagggga ccaaattata accatgtatc tagaggatgg 12360tttcaaatta ataaaacact tagaaccctt atgtgtcagt tgtatacaaa cgtacggtat 12420ttttacgccg aggaagtact ggtttcaatc tcagatgatt aaatcatatt atgatgaact 12480tcaaagtctt aacctaaaac ttcagattcc agataatagg actgaatgtg cacagaactt 12540tattaaaacc ataattcagg caaaactgac tcctcaacaa tactgtgaat tgttctcttt 12600acaaaaacat tggggtcacc cagttttata caatgatgtt gcactagaca aagtgaagaa 12660acatgcccaa tcaacaaaaa ttttaaaacc taaggtcatg tttgaaactt tttgtgtctt 12720taagtttata gtggcaaaaa atcattatca ctctcaagga tcgtggtaca aaaccacaca 12780tgatctacat ttgaccccat atctacggca gcacattgtg tcaaattcat ttccatcgca 12840agccgagatt tatcagcacc tctgggaatg gtactttgta gagcatgaac ctcttttttc 12900tacaaaaata ataagtgatt tgagtatttt cataaaagat agagctactg ctgtaaatcg 12960agagtgttgg gacagcgttt ttgataggag tgtgctagga tacaatcccc ctgtcagatt 13020tcaatcaaag agggtacctg agcaattctt aggtcaagca gatttctctt taaatcaaat 13080attggatttt gctgagaaat tagagtatct agctccttca tatagaaact tttccttttc 13140attaaaggaa aaagaattga atattgggag aacattcggg aagttgccct atcgtgtcag 13200aaatgtccaa acacttgcag aagccttatt agcagatgga ctagcgaagg cattccctag 13260taatatgatg gttgttactg agagagagca gaaagaagct ctattgcatc aagcctcttg 13320gcaccataat tcagcaagta taggggagaa tgccatagta agaggtgcaa gttttgttac 13380tgatcttgag aaatacaatc ttgcctttag atatgagttt acacgacatt tcatagacta 13440ctgtaatcga tgttacggtg ttaaaaattt attcgactgg atgcatttct taataccact 13500atgctatatg catgtcagtg acttttatag ccctcctcac tgcgtgacag aaaataaccg 13560aaataaccca cctgattgtg ccaatgctta tcattatcat ttaggaggta tagaaggact 13620acaacagaaa ttgtggacat gtatatcatg tgctcaaatc acgcttgtag aactgaaaac 13680taagttaaaa ttgaaatcca gtgtcatggg tgataatcaa tgtataacaa ctctaagtct 13740tttccctatt gatgctccca atgattatca agaaaatgag gcagaattaa atgctgcacg 13800agttgctgtt gaattagcta ttactacagg ctatagtggg atattcttaa aacctgaaga 13860gacatttgtg cattcagggt ttatttactt tggtaaaaag caatacctaa atggtgttca 13920attaccacaa tcattgaaga caatggcaag gtgtggacct ttatcagatt ccattttcga 13980cgatcttcaa ggttcactag ccagtattgg cacatcattt gagagagggg caagcgagac 14040acggcatatt tttccaagtc gttggatagc tgcatttcat tccatgttag ccgtaaattt 14100gttaaatcag aatcacctcg gatttcccct aggatttagt attgatgtat cctgttttaa 14160aaagcctctt actttctcgg agaaattaat tgctttgatc acacctcaag tgctaggggg 14220attatctttc ttaaacccgg agaaattatt ctatcggaac atcagtgatc ctcttacttc 14280agggttattt caactcagga atgcattaga gttccttaga aaggaagagt tgttctacat 14340cttgattgct aaaaaacccg gtttagctga tgcttcagat ttcgttatga atccattagg 14400tttaaatgtg ccaggatcta gggaaataat aacgtttctc aggcaaacag ttcgtgaaaa 14460cataacaatt acatcacaaa atagaataat aaattctctt tttcacatag gttccgactt 14520agaggatcaa agagtatgtg aatggctttt atcatcaaac cctgtaatga gccgatttgc 14580tgctgatatt ttttcacgaa cacctagtgg aaaaagactt caagttttag gttacttgga 14640aggaaccaga acgttattag cttcccgcac gatcagttta accactgagg gtacaatgtt 14700gatgagatta agagagttaa ctaagagtcg atggaagagt tggttttctt acattgatgc 14760attagatgat gatttgtctg agtctcttga aaagttcata tgcactgttg atgtggctaa 14820tttcttgaga gcatattcat ggtcagatgt cttgaaagga aaaaggttaa ttggtgccac 14880actaccatgt ttactggagc aattcaatgt aaagtgggtc aacttgtctg aagacttaaa 14940ggagcaattt aagctatctt cagatctggg atcacctacg gatttattgc agtacgattg 15000caatggactg cattcaaagg gggccgataa cgcagaatta aattatgtga gttgtgccct 15060tgaccggaaa attgttcaaa agcatccatc tgacaatcgc ctggcatgga caataggaaa 15120tcgagcaccg tatatagggt cacgaacaga agataaaatt ggttaccctc ctttaagagt 15180aaactgccca tcggcagccc ttaaagaagc tattgaaatg gtctctagac tattgtgggt 15240gactcaaggc accgcagatc gagaaaaatt gctcattcct cttctcaatt caagagttaa 15300tttagactat cagacagtgc tcaacttcct gcccactcac tactcaggca atatagttca 15360cagatacaat gatcaatatg

gacaacactc ctttatggca aatagaatga gcaatacatc 15420cactcgtgca atcatatcaa ctaacacact agggaaatat gctggagggg gtcaagctgc 15480tgttgatagt aatataatct tccagaacac tattaattta ggtgttgcag ttttggacat 15540tacattgtct ctttctaaat tgtcatcaac atcaaatgtt tctttccgtt taatgttaag 15600taaatgttgc acacggcatg taccgtctga gtatttattc tttgataaac ctttagatgt 15660ggatttgaac aagtacatgg acaacgagtt agtttacgat aatgatcctc tctgtagtgg 15720aataaaggga agattaggta gagtatcaag atcaacactc tcattgagtc taaatgtaag 15780cgatattgga tcttacgact ttccgactat tgctgcgtgg actttaggag agacaattat 15840tggaagtatt ttttctgatg aatcttctca aagtacagac cctataagtt caggctgtac 15900aaaaactttt gtaacacact ttcttgtgta tccagttgag agtatctttt atgcctttgg 15960agctaatcta atagtggaaa gtttaagttt aagcaggatc aattcaatca agagcctctc 16020agatttaaca tttcttatat catccacaat cagaaatttg tcacacagat cacttcgaat 16080tcttcaatct actttccgac atgaattggt attaactaga ctagctcatc atataccatt 16140gatttcctta atgctaggag gttctgcggg tgagaaaagt tcgtcggatg ctgtccgact 16200atttcttacg gcaagttatc aaaattttat caataatttc agttgtttga tgagaaagaa 16260ccaatcacca ttaccagttt ggctttattt ccctagtgaa gggcaacaac taaaacctat 16320tttaaaaatt ttgcaaaggt tatcatgttt attaacaact aaaaaggttc aaaatcacag 16380acctgtagct gatacttgtt ttttgactga taatttttgg gtctatccaa gcaaatcaac 16440gagaactaat cattattatg caagtcttaa ttattggaga gacaaagcta ataagattaa 16500gaatacttca ttttcacatt tgataaacta ttcattttct gaaccctctc tacatgcgag 16560ctctatctct tctagtcaag aagtggtcaa tttaaaacac accagtcgtt tagatgaaac 16620acctaatatg agtgaaaggg ctcaatcaac aaatcatgag ccaacagctt tacaagaggt 16680gtgcactgag ataccctact cggaacaaga tccagccaaa agttatttgc tgttagagaa 16740tactagattc agggatgatc agaaaatatt aagacatgat cagaaagctg agaggggtga 16800acctctttca ttgcaagtgt cttctagggg ttgcctgcag gctcttactt gccctcatca 16860cccctcccca tctcaaacca ccacagaacc actaagcatg cttaggaatt gtgacgccat 16920aaaagcagcc ttacgttctg agacgaatga tccccgtctt atgagcagta tccttgatat 16980gagatcattg aaaactccca tgagaataga atctcgaaac acgagtctat tgcaaccttc 17040tgagtgtctg tcaacttcta agggaaaatc tgtactgtct agagaacagg cttcatacct 17100gtatgttgat tgcagtaata tctcttctat ttctctggat tcaggttttc gaaatatgtc 17160tgatagaaat caagtccaaa tgctaataaa tacttacaaa cgtgacttat acacttgttt 17220tgatagtaac caattctgca ggtttacagg ggtcgtttca tcaatgcatt ataagcttta 17280tgatcttttg ccagcaggca aactcggaaa ggcaatctgc ctagccgaag gggaagggag 17340tggcgctcga ctactcttga agtggaagga gacagattat ttattcttca atactttggc 17400cacagattca caacaggaag cagaaatttt gagtggtcga gttattccaa ggatgttgta 17460taacatagat aagctaagtg ttttacttga atccagaaaa ttaatcttga ataatctaac 17520tattcaaatc acggatatta caaacccact atggctggac tctgtcatac aatacctacc 17580tgaagatagt gacattctaa caatggatgc agagaccact aaagaagaga caagagagca 17640actctataaa actatcataa atatttgggc acgtacttct cctaatatcc ctaaaaccag 17700catcattaaa gtgtttttat tagattatgg gggaaccttg ttcttaatga agaatgctat 17760tcaatattat ggacaagttc aacttaagaa accatatagt tcaaatgcaa aaaattcaga 17820atggtactta tgttgtggaa aacgaagagt tcaacgactc cgagttgatt ttccagacca 17880agtaggaata ttcttgatct gtaaagcaat gtcacgtcag aggcaagcaa ttccttactg 17940gctaaagcac atagaaaaga attaccctgc ttcattgcac gagttcttta taactttagg 18000ttttccttct ttagagtcat ctttctgcca tcgctacacc attccgttca ctgagggaac 18060ggctctcttt cacaaggtcc agtcttatgt ccgacaaggt agacaacacc tacactctct 18120tatgttagat tacgaaaata attcacccct cctagatctg agaaatcact tcatatgctc 18180attgagggga aagataacca agtattacaa tgacatattg aaattaaatc tagtagtgag 18240agcagtagaa agagggaaaa attggtcgca actcgttgag tcccttccta atatgcactc 18300agtatgcata acacatgttg atcacgaatg tattggctgt gagagacggt tattacttaa 18360attggacttt gtcagaaata caaagatagc agaacaaaag ttactcaata gggtaattgg 18420gtatattcta ttcttccctt ttggtttctc cagacccaag tgactacaga tatattcttc 18480aataaaggaa gctcagtcta actcacagaa ataatctact tcaaaacaag gatcacccat 18540tttggaacat tgtataagaa actgcaagac aaataataag gaaaggatac tactgtataa 18600cttgttatat ccaaaaggat cttgggtcat tttaagcatg atgcaaataa aaaatcgtct 18660acatagccga actgaccgct cagtacttat tcacaatcat gctaactttt agtttttaat 18720tgtgcaaaaa ttactaagaa taattaatat tgatattaaa acattaaatg gacatttgag 18780ttttatgcct agaataatat aaagaaattt aagagacatt tagatatatc agttgaattg 18840atttatgaca catagtgcat catgaataca aagagaaaaa ttgttgcaat tcaggaatat 18900cttatttaaa tgtattagag agaaagtcag attattatca aaatcaagca aaatacaata 18960ggttttttca aagaataggt ggtaaagcct tatggttatt ttttaaagat gtcaatgtga 19020atttttatta agaaaaagta atgcatgaaa ttaaaaaatt aaagaacttt gatataagta 19080acacaaaaca ctcttcatct ttttagtgtg tcca 1911420281PRTLake Victoria marburgvirus 20Met Gln Gln Pro Arg Gly Arg Ser Arg Asn Arg Ser His Gln Val Ala1 5 10 15 Leu Ser Thr Tyr His Glu Asn Gln Leu Pro Ser Lys Pro Gln Tyr Ile 20 25 30 Asn His His Pro Arg Ala Arg Ser Met Ser Ser Thr Arg Ser Ser Thr 35 40 45 Glu Gly Ser Pro Thr Asn His Ala Ser Arg Ala Arg Pro Leu Ser Thr 50 55 60 Phe Asn Leu Ser Lys Pro Pro Pro Pro Pro Lys Asp Met Cys Arg Asn65 70 75 80 Met Lys Ile Gly Leu Pro Cys Thr Asp Pro Ala Cys Asn Arg Asp His 85 90 95 Asp Leu Asp Asn Leu Thr Asn Arg Glu Leu Leu Leu Leu Met Ala Arg 100 105 110 Lys Met Leu Pro Asn Thr Asp Lys Ala Phe Lys Ser Gln Gln Asp Cys 115 120 125 Gly Ser Pro Ser Leu Ser Lys Gly Leu Ser Lys Asp Lys Gln Glu Gln 130 135 140 Ala Lys Asp Val Leu Thr Leu Glu Asn Leu Gly His Ile Leu Asn Tyr145 150 155 160 Leu His Arg Ser Glu Ile Gly Lys Leu Asp Glu Thr Ser Leu Arg Ala 165 170 175 Ala Leu Ser Leu Thr Cys Ala Gly Ile Arg Lys Thr Asn Arg Ser Leu 180 185 190 Ile Asn Thr Met Thr Glu Leu His Ile Asn His Glu Asn Leu Pro Gln 195 200 205 Asp Gln Asn Gly Val Ile Lys Gln Thr Tyr Thr Gly Ile His Leu Asp 210 215 220 Lys Gly Gly Gln Phe Glu Ala Ala Leu Trp Gln Gly Trp Asp Lys Lys225 230 235 240 Ser Ile Ser Leu Phe Val Gln Ala Ala Leu Tyr Val Met Asn Asn Ile 245 250 255 Pro Cys Glu Ser Ser Ile Ser Val Gln Ala Ser Tyr Asp His Phe Ile 260 265 270 Leu Pro Arg Asn Gln Gly Glu Arg Gln 275 280 2118890DNAReston ebolavirus 21gggacacaca aaagaaaaag gttttttaag attttttgtg tgcgagtaac tatgaggaag 60attaacagtt ttcctcagtt taaggtatac actgaaattg agattgagat tctcctcttt 120gctattctgt aactttccct ggttgtgaca attgaatcag ttttatctat taccaattac 180catcaacatg gtatgtctag tgatcttggg actcttcttc atctggtttt tcctagagct 240ctgaatctat tttgtgagaa gttcatccaa acgacccagt gtctgaaaat acaagaggtt 300cccctttccg tcaagtttaa ggggttgttt tgattgtgtg tagattttat aatcctagag 360tgccaaggag ttgcgtgtca tcattaattg ggaagatcaa ggaaacaatt tgttccaata 420atatcgtaca tcttgactaa gtcgaacaag gggaagtcga tatggatcgt gggaccagaa 480gaatctgggt gtcgcaaaat caaggtgata ctgatttaga ttatcataaa attttgacag 540ctggccttac tgttcaacag ggaattgtca ggcagaaaat aatttctgta tatcttgttg 600ataacttgga ggctatgtgt caattggtaa tacaagcctt tgaggccgga attgatttcc 660aagaaaatgc cgacagcttc cttctgatgc tttgcctaca tcatgcttac caaggtgact 720ataaattgtt cttggagagc aatgctgtac agtatttgga aggtcatgga ttcaaatttg 780agctccggaa gaaggacggt gtcaatcggc tcgaggaatt gcttcctgct gcaacgagtg 840gaaaaaacat caggcgtacg ttggccgcac tgcctgaaga ggagactaca gaagcaaatg 900cagggcaatt tctctcattt gcgagtttgt ttcttcccaa actggttgtg ggagagaagg 960cttgcttgga aaaagtccag cgacaaattc aggttcatgc agaacagggt ttaattcaat 1020atcccactgc atggcaatca gttggacaca tgatggtaat cttcagattg atgaggacta 1080atttcttgat taaatattta ctgatccacc agggtatgca tatggtagct ggccacgatg 1140ccaatgatgc tgtcattgct aattcagttg ctcaggctcg cttttcagga ctcctaattg 1200tcaaaaccgt tcttgatcat attctgcaga aaaccgacca aggagtaaga cttcaccctt 1260tggcccgaac agccaaagtg cgtaatgagg ttaatgcatt taaggccgcc ctaagctcac 1320ttgctaagca tggggagtat gccccttttg ctcgccttct caatctctcg ggagttaaca 1380acctagaaca tggtctctac ccacagttat cagcaattgc tcttggagtt gccacagcac 1440atggtagcac ccttgcagga gttaatgttg gtgagcagta tcagcagctt agagaggctg 1500ccactgaagc tgagaagcaa ctccaacaat atgctgagtc cagagaactc gacagcctag 1560gcctagacga tcaggaaaga agaatactaa tgaacttcca tcagaagaaa aatgaaatta 1620gtttccagca gaccaatgca atggtaaccc ttaggaaaga gcgactggct aaattaacag 1680aagctataac gctggcctca agacctaacc tcgggtctag acaagacgac gacaatgaaa 1740taccgttccc tgggcctata agcaacaacc cagaccaaga tcatctggag gatgatccta 1800gagactccag agacactatc attcctaata gtgcaattga ccccgaggat ggtgattttg 1860aaaattacaa tggctatcat gatgatgaag ttgggacggc aggtgacttg gtcttgttcg 1920atcttgacga tcatgaggat gacaataaag cttttgagct acaggacagc tcaccacaat 1980cccaaaggga aatagagaga gaaagattaa ttcatccacc cccaggcaac aacaaggacg 2040acaatcgggc ctcagacaac aatcaacaat cagcagattc tgaggaacaa gaaggtcaat 2100acaacaggca ccgaggccca gaacgtacga ccgccaatcg aagactctca ccagtgcacg 2160aagaggacac ccctatagat caaggcgatg atgatccctc aagcccacct ccgctggaat 2220ctgatgatga cgatgcatca agtagccaac aagatcccga ttatacagct gttgcccctc 2280ctgctcctgt ataccgcagt gcagaagccc acgagcctcc ccacaaatcc tcgaacgagc 2340cagctgaaac atcacaattg aatgaagacc ctgatatcgg tcaatcaaag tctatgcaaa 2400aattaggaga gacatatcac catctgctga gaactcaagg tccatttgaa gctatcaatt 2460attatcacat gatgaaggat gagccggtaa tatttagcac tgatgatggg aaggaataca 2520cctacccgga ttcacttgag gaagcctatc ctccatggct caccgagaaa gaacgactgg 2580acaatgaaaa tcgatacatt tacataaata atcaacagtt cttctggcct gtcatgagtc 2640ccagagacaa atttcttgca atcttgcagc accatcagta accacagcac aaagcgcggt 2700ccacttcgta aagctaaata cacttaaagc ttgaccgatt catctacaaa aactaatcca 2760ttataactta ttagtgctac ttttctataa gtgattctca atctaaggcc attaagagtt 2820taagcaatat acatatacac ttacaccggt ctatccaaga tgtggctcaa tgttcttaat 2880ttgaacatag tcataagggg ataaataata ctttatattt ctgattgtgg actgacccat 2940tctgcttaaa atgcttcgcc cattaaaaat gtgatctaat agatagccct gactagacca 3000attaagaaaa acatttgatg aagattaaaa ccttcatcgc cagtaaatga ttatattgtc 3060tgtaggcagg tgtttactcc accttaaagt cggaaatatc ctaccttagg accattgtta 3120agaggtgcat aggcattacc atccttgaga acatgtataa tgataaattg aagatatgtt 3180caggcccaga aacaactgga tggatttctg agcaactaat gacaggtaag attccagtaa 3240ctgatatatt cattgatatt gataacaagc cagatcaaat ggaagtccgg ctcaaaccat 3300catcaaggag ctcaaccaga acttgtacaa gtagcagtca gacggaggtc aactatgtac 3360ctctccttaa aaaggttgag gatacattaa ctatgctagt gagtgcaacc agtcgtcaga 3420atgctgcaat cgaggccctt gaaaaccgcc tcagcacact tgagagtagc ttaaagccaa 3480tccaagacat gggtaaagtg atttcatcat tgaatcgcag ttgtgccgaa atggtggcaa 3540aatatgatct tctagttatg acaactggac gggctacttc aaccgcagct gcagtagatg 3600cgtactggaa agagcacaaa cagccaccac cagggccagc gttgtatgaa gagaatgcgc 3660ttaaaggaaa aatcgatgat ccaaacagct atgtaccaga tgctgtgcag gaggcttaca 3720agaaccttga cagtacatcg accctgaccg aggaaaattt tgggaaacct tatatatctg 3780ctaaagatct gaaggagatc atgtatgatc atctacctgg ttttgggact gcctttcacc 3840aacttgttca agtgatttgt aaaataggaa aggataacaa cctcttggac acaatccatg 3900ctgagttcca ggcaagtcta gcagatggtg actctcccca atgtgcactc atacagataa 3960ccaaaagagt cccaatcttt caggatgtgc cgcccccgac aatccacatt agatcccgtg 4020gtgatatccc acgagcatgc caaaagagtc tccgaccagc accaccatca cccaaaattg 4080atcgtggttg ggtttgtttg tttaagatgc aagatggtaa aacgcttgga cttaagatct 4140aaggatcaag atttatttaa caaggcaagc cacaacctta gatagaacct cagccagact 4200attgaactat tgacgctgtt gatgataata tataattaat ggtcatattt gaatatgaca 4260acatcttgct tcttgttttg ccttgtatct ctttgagttg gaagatcatt ccaaacttac 4320aaacatgcac aagatgttat ggtttagcaa agaattgata ggagtactgg tatataatgt 4380aaatataaca agtgatgaag attaagaaaa accagtcggt attttccaga cttggcattt 4440cttatcttca tcttctaaag tgagatattt tatcatcaaa aaatgagacg cggagtgtta 4500ccaacggctc ctccagcata taatgatatt gcatactcta tgagcatact cccaacccga 4560ccaagtgtca tagtcaatga gaccaaatca gatgtactgg cagtgccagg agcagatgtt 4620ccatcaaact ccatgagacc agtggctgat gataacattg atcactcaag ccatactcca 4680agcggagtag cttctgcctt tatattggaa gctaaagtga atgtaatttc gggaacaaaa 4740gtcctgatga agcaaatacc tatttggctt ccactgggtg tagctgatca gaagatatac 4800agctttgatt caacaacagc cgcaattatg ttggcttcct acacagtgac acacttcggg 4860aagatatcta acccgctggt acgtgtcaac aggctaggcc caggaatacc cgatcatccg 4920ctacgactcc taaggttggg caatcaggca ttccttcaag agtttgttct tccaccagtc 4980cagcttcccc agtatttcac atttgatcta acagctctaa agctcatcac tcaaccattg 5040ccagctgcaa cctggacaga cgaaactcca gcaggagcag tcaatgctct tcgtcctggg 5100ctctcactcc atcccaagct tcgtccaatt cttctaccgg ggaagatagg aaagaaaggt 5160catgcttcag acttaacatc acctgacaaa attcaaacaa tcatgaatgc aataccggac 5220ctcaaaattg tcccgattga tccaatcaag aacatagttg gaattgaggt tccagaatta 5280ctagttcaaa ggctgaccgg caaaaaacca caacccaaaa atggccaacc aattattcca 5340gttcttcttc cgaaatatgt tggacttgat cctatatcgc caggggactt aactatggtt 5400atcacccagg attgtgattc atgccactct ccagccagcc atccgtatca catggacaag 5460caggatagtt accaataatt taaattccat tcgagctatt attctgctag taattccgac 5520gggatcaata gactaaaaat ctgattgtat agaattataa aagaatcaag cagaggcaac 5580agactcacag cttacgccta gatgactaat attaaggagt tttttaatct aattttccag 5640tcttaagtaa taatcatttc ttttgtaatt aattatgcat ttgttaactt atcggtgcga 5700gatttccttg agaacccggc ggggcttcta ctatctgtag taaccagaag agaagttcaa 5760cccagtcaaa actaaaccaa gcaatattct gaatgctcta tagtctattc taatcagagg 5820tataacaatg gctaagattt caatgactcg ttaacaatcg ctagtaattt taatctccag 5880attaagaaaa agatatacga tgaagattaa ggcgacaacg agccgaaact tcatctcttt 5940taaagatcta acattatctg ttccaaagtc atacaaggac acattcaaat cagggattgt 6000aagctgctat ttcttacctc cccaaatcac ctatacaaca tggggtcagg atatcaactt 6060ctccaattgc ctcgggaacg ttttcgtaaa acttcgttct tagtatgggt aatcatcctc 6120ttccagcgag caatctccat gccgcttggt atagtgacaa atagcactct caaagcaaca 6180gaaattgatc aattggtttg tcgggacaaa ctgtcatcaa ccagtcagct caagtctgtg 6240gggctgaatc tggaaggaaa tggaattgca accgatgtcc catcagcaac aaaacgctgg 6300ggattccgtt caggtgtgcc tcccaaggtg gtcagctatg aagccggaga atgggcagaa 6360aattgctaca atctggagat caaaaagtca gacggaagtg agtgcctccc tctccctccc 6420gacggtgtac ggggattccc tagatgtcgc tatgtccaca aagttcaagg aacaggtcct 6480tgtcccggtg acttagcttt ccataaaaat ggggcttttt tcttgtatga tagattggcc 6540tcaactgtca tctaccgtgg gacaactttt gctgaaggtg tcatagcttt tttaattctg 6600tcagagccca agaagcattt ttggaaggct acaccagctc atgaaccggt gaacacaaca 6660gatgattcca caagctacta catgaccctg acactcagct acgagatgtc aaattttgga 6720ggcgaggaaa gtaacaccct ttttaaggta gacaaccaca catatgtgca actagatcgt 6780ccacacactc cgcagttcct tgttcagctc aatgaaacac ttcgaagaaa taatcgcctt 6840agcaacagta cagggagatt gacttggaca gtggatccca aaattgaacc agatgttggt 6900gagtgggcct tctgggaaac taaaaaaact tttcccaaca acttcatgga gaaaacttgc 6960atttccaaat tctatcaacc cacaccaaca actcctcaga tcagagcccg gcgggaactg 7020tccaaggaaa aattagctac cacccaccca ccaacaactc cgagctggtt ccaacggatt 7080cccctccagt ggtttcagtg ctcactgcag gacggacaga ggaaatgtcg acccaaggtc 7140taactaacgg agagacaatc acaggtttca ccgcgaaccc aatgacaacc accattgccc 7200caagtccaac catgacaagc gaggttgata acaatgtacc aagtgaacaa ccgaacaaca 7260cagcatccat tgaagactcc cccccatcgg caagcaacga gacaattgac cactccgaaa 7320tgaattcgat ccaaggctcg aacaactccg cccagagccc acagaccaag gccacgccag 7380cgcccacagc atccccgatg accctggacc cgcaagagac ggccaacatc agcaaaccag 7440gaaccagccc aggaagcgca gccggaccaa gtcagcccgg actcactata aatacaataa 7500gtaaggtagc tgattcactg agtcccacca ggaaacaaaa gcgatcggtt cgacaaaaca 7560ccgctaataa atgtaaccca gatcttcact attggacagc tgttgatgag ggggcagcag 7620caggattggc atggattcca tattttggac ctgcagcaga aggcatctac attgagggtg 7680taatgcataa tcagaatggg cttatttgcg ggctacgtca gctagccaat gaaactaccc 7740aggctcttca attatttctg cgggccacaa cagaactgag gacttactca cttcttaaca 7800gaaaagctat tgattttctt cttcaacgat ggggaggtac ctgtcgaatc ctaggaccat 7860cttgttgcat tgagccacat gattggacaa aaaatattac tgatgaaatt aaccaaatta 7920aacatgactt tattgacaat cccctaccag accacggaga tgatcttaat ctatggacag 7980gttggagaca atggatcccg gctggaattg ggattattgg agttataatt gctataatag 8040ccctactttg tatatgtaag attttgtgtt gatttattct gagatctgag agaaaaaaat 8100ctcagggtta ctctaaggag aaatattatt tttaaaattt acttaaatgc tgaccactta 8160tcttaaatga gcaattaata atatgttttt ctgcttcttt gcttgattta caatatgata 8220tttctcttaa taatgattaa tatattaaga aaaacttatg acgaagatta aaggggagga 8280tcgttaacgg gaaaatctcc catctcgttc gtcgaagcca cgttggtggt gcttgcagct 8340gagaacaact ccagagattg taggtagaaa ggaccagcat ttataggtag gggtcagaaa 8400gcaacaatag ccataaaagg agagcctgac attgctattt aatatcctag aacctgattt 8460ctaggttcta gttgtacaat ccggatgatg gagcattcaa gagaacgggg tagatctagc 8520aacatgcgac ataatagccg ggaaccatac gaaaatccat caaggtctcg ctcattatct 8580cgggacccta atcaggttga tcgtaggcag cctcgaagtg catcccaaat tcgtgttccg 8640aatctgttcc atcggaaaaa gactgatgca ctcatagttc ctccggctcc caaagatata 8700tgcccaacac tcaaaaaagg attcctctgc gatagtaaat tttgcaaaaa agatcaccaa 8760ttggatagct taaatgatca tgaattacta ctgctaattg caagaagaac atgtggaatt 8820atcgagagca attcgcagat tacatcccca aaagatatgc ggttagcgaa tccaacagct 8880gaagacttct cacaaggtaa tagtcctaaa ttaacacttg cagtccttct tcaaattgct 8940gaacattggg caaccagaga cctaaggcaa attgaggact ctaaacttag agctctttta 9000accctttgtg ccgtattaac aaggaaattt tctaaatccc aactgggtct tctatgtgag 9060acccacctac ggcatgaggg cctcggacag gaccaagctg attctgtatt agaggtctac 9120caaagactcc acagtgataa aggagggaat tttgaggctg ccctgtggca acaatgggac 9180cgacagtcgt taataatgtt catctctgct tttctcaaca ttgctctcca gacaccttgt 9240gaaagttcta gtgtcgtagt ctcaggtctt gccacattgt acccagcaca agacaattct 9300acaccgtccg aggcaactaa tgataccacc tggtcaagta cagttgaata gaaaaccact 9360ggagctattt ttccacgatt gctctcagtc aataaattaa

tatagatata atacgacttc 9420ggtgtgcaat tgtcaagggt tccatttggt aataatgatt cttaaaacaa tctactatcg 9480taattatcga tggatctacc ctatttgacg gtacatgact tgaatgtaat aaggtaagtt 9540ggtatctgag gtattttgtc tagagtatac tcaaaatcgt atgtctagca aattatcaat 9600agcaaagtta aattctccta acctcatatt ttgatcaagt aatcatgatt ttatggtaat 9660tctttgcaga ttatcggttt aatctttatt aagaaaaaat catgattgta gacaatttac 9720tggtagtccc tgggtatcca agtttatgaa cagagctaga gagaatttgc tacttccgag 9780gtataacttt attatttgct acttcgaatg cctaaaacca gtaatgcagg atgaagatta 9840attgcggagg aatcaggaat tcaactttag ttccttaagg cctcgtctga atcttcatca 9900gttagtaagt tcttttatag aagtcattag cttctaaggt gattatattt tagtattaaa 9960ttttgttaat tgcttgctat aaagttgaaa tgtctaatgc ttaaatgaac atttctttga 10020agctgacata cgaatacatc atatcatatg aaaacatcgc aattagagcg tccttgaagt 10080ctggcattga cagtcaccag gctgttctca gtagtctgtc cttggaagct cttggggaga 10140caagaagagg tcccagagag tcccaacagg ttggcataag gtcattaaca ccagcatagt 10200cagctcgatc aagactgtaa gcgagtcgat tgcaactaaa aagattattt cttgttgttt 10260aaacaaattc cttttgtgtg agacaccctc aaggcacaag atggctaaag ccacaggccg 10320atacaatctc gtgcccccaa agaaagatat ggaaaaggga gtgattttta gtgatctttg 10380taatttcttg attactcaaa ccctgcaagg ttggaaggtt tattgggcag gaattgagtt 10440tgatgtaagt caaaaaggca tggctcttct gacaagactc aaaacaaatg actttgctcc 10500tgcctgggcg atgacaagaa atctcttccc acatctgttc caaaacccaa attcggttat 10560tcaatctccc atctgggctt tgagggtgat tttggcagcc ggattgcagg atcagttgtt 10620agaccattca ttggttgagc cattgacagg ggctctcggt ctaatttctg attggctcct 10680aactacaacg tcaacacatt tcaatcttcg tactagaagc gtaaaggacc agcttagtct 10740tcgtatgtta tctttgatca ggtcaaacat cttgcagttc atcaacaagc ttgacgccct 10800gcatgttgtc aattacaatg gtttactcag tagtattgag atcgggactt ctacacacac 10860aatcattata actcgtacaa atatgggttt tctcgtggaa gttcaggagc ctgacaaatc 10920agctatgaat tctaagcgcc caggaccagt caagttctca ttacttcatg agtctgcctt 10980caaacctttc actcgtgttc cacaatctgg gatgcaatca ttaataatgg agttcaacag 11040tttgttggca atttaacaag gtgatcttaa aataagtaca tgaatgagaa ttagttgtgg 11100gtcttaccta gcattgttga gttagctatc taatctattt tcactaattg cattgagcac 11160tgctagtagg tttgcaccac gttaaagatt cagagtgtat gaattgtgca gatttaaact 11220tgggttttgc cttatgcttc acaggtggtc tttttaaaat ggagattatc agcatttctt 11280caatgggagg agttagcaat cagaaattgg agataaatgg acatcgggat agaacaatgc 11340ctaactattg ggcggctttc atttttaaat gtgtatataa ccaatctttt cctatctttg 11400cttatattgg tgtaacttta ctttaataac atgtcaatgc tatactgtta agagaaggtc 11460tgaggaagat taagaaaaag gtctcgtgtt cacttggttg ccgtcaagta tcctgtggtt 11520tttttctacc taacttcctc atgccatatg gctacccagc atacccagta cccggatgca 11580cgtttatctt cacctatagt cctggatcaa tgtgatttgg taactcgagc atgtgggtta 11640tattcatctt attctctaaa tcctcagcta aggcaatgta aattaccaaa acatatatat 11700cgacttaagt tcgacacaat agtatccaaa ttcctaagtg atacacctgt agcaacactg 11760ccgatagact atttagtacc aattctcctg cgttccctaa cggggcacgg tgataggccg 11820ttgaccccga cttgtaatca attccttgat ggaattatta attacactct tcatgatgca 11880gcctttcttg attactatct caaggcaaca ggtgcacagg accatttgac aaacattaca 11940actagagaga agcttaaaaa cgaaattcta aacaatgatt atgtccatca attgttcttc 12000tggcatgacc tgtctatttt ggctcgacgt gggcgtctga atcgcgggaa caaccgttca 12060acctggtttg ttcatgatga attcattgat attttaggat atggcgatta tattttttgg 12120aaaatacctt tatcattatt accagttact atagacgggg tcccacacgc ggcaactgac 12180tggtatcaac cgactctttt taaagaatcc atcctagggc acagccaaat cctatctgtg 12240tcgacagctg aaatactaat tatgtgtaaa gatattatca cctgtaggtt taatacatca 12300ctgattgcat ccattgcaaa attagaggat gtagatgtgt ctgattatcc tgacccgagt 12360gatattctta agatatacaa tgctggagac tatgtaatat ctattcttgg ctcagaaggt 12420tataagataa taaagtacct tgaaccactt tgtttggcca aaatccaact ttgctctaaa 12480ttcacagaaa gaaaaggtcg tttcctcaca cagatgcatt tatcagtaat aaatgatctt 12540cgggagttga tttctaaccg caggttaaag gactatcagc aagagaagat tagggatttt 12600cacaaaatat tattacaatt gcaattatct cctcaacagt tttgtgaatt attctctgtt 12660caaaaacatt gggggcatcc aattttacat agtgagaaag ctatacaaaa agtaaaacgg 12720catgcaacca tccttaaggc tctcagacct aatgtcattt ttgagacata ttgtgtattc 12780aagtacaata ttgccaagca ctatttcgac agccaaggaa cttggtacag tgtaatctca 12840gacaggaatt taactccagg actcaactcc ttcataaaac gtaatcactt tccttcacta 12900cccatgatta aggatcttct atgggaattc tatcatctta atcaccctcc gttattctct 12960acaaaggtga ttagtgactt aagtattttc atcaaggata gggccacagc tgttgaacag 13020acatgttggg atgcagtctt tgaacccaat gtgctaggtt acaatcctcc aaacaaattc 13080tccactaaaa gggtgccgga acaatttcta gaacaggagg atttttcaat cgaaagtgtc 13140ctgaattatg cacaggaatt acattattta ttaccacaga ataggaattt ttccttttct 13200cttaaagaaa aagaattaaa tattggacga acatttggta agctaccata tctcacacgg 13260aatgtccaaa ctttatgtga ggctctgtta gcagatggac tggctaaggc cttccccagt 13320aacatgatgg tagtaactga acgtgaacaa aaagagagcc ttcttcatca ggcatcatgg 13380caccacacca gtgatgattt tggagagaat gctaccgttc gagggagtag ttttgtaact 13440gatttagaga agtacaatct tgcatttcgc tatgagttca ctgcaccatt tattgagtac 13500tgcaaccatt gctatggtgt gcgtaatgtc tttaattgga tgcattattt aatcccgcag 13560tgttacatgc atgtaagtga ttattataat ccgcctcaca atgttaatct tagcaatcga 13620gaatatcctc ctgaaggccc gagttcgtac cgagggcact taggaggcat agagggatta 13680caacaaaaac tgtggacgag tatatcctgt gcacaaatct ccttagtgga aattaaaact 13740ggttttaagt tacgatcagc ggtcatggga gacaatcagt gtataaccgt attgtctgtt 13800tttccacttg aaacagaccc tgaagagcag gagcaaagcg ccgaagacaa tgctgcaaga 13860gtagcagcaa gtcttgcaaa agtaaccagt gcatgtggga tctttcttaa accagaagag 13920acattcgtac actcaggttt catttatttc ggaaaaaaac aatatctcaa tggtgtacaa 13980ttaccgcaat cactcaaaac agcagcaaga atggcgccac tctctgatgc tatattcgat 14040gatctacaag gaacacttgc cagtattgga actgccttcg aacgtgctat atcggaaacg 14100cgacatatcc tcccatgtcg tattgtagca gctttccata cgtatttcgc cgttcggatt 14160ttacaatatc accatcttgg atttaataaa ggcatcgatt tagggcagtt gtcacttagt 14220aaaccattag actatgggac tattactcta acattggcgg ttccacaagt ccttggggga 14280ttgtcttttc taaatccaga aaagtgtttt tatcgaaact tcggagatcc tgtgacttct 14340ggacttttcc agctacgggt gtacctagaa atggttaaca tgaaagacct attttatcca 14400ttaatatcga aaaatccagg aaattgtagt gccattgatt ttgtcttaaa tccatccgga 14460ttaaatgttc caggatcaca agacttgaca tcctttttgc gacagatcgt taggcgtagt 14520attacactaa ctgcaagaaa taagttaatt aacactctct tccatgcctc tgctgatttg 14580gaagatgaga tggtttgtaa atggctcctt tcatcaaacc ctgtcatgag tcgctttgca 14640gcggatattt tttccaggac acctagtggt aaacgtctcc aaatattagg ttatcttgaa 14700gggaccagga ctctattggc ctccaaaatc ataaacaaca acagtgagac acctgtactt 14760gataagctga ggaagatcac cctacaaaga tggaatctgt ggttcagtta tttggaccat 14820tgtgaccaat tactagcaga tgctctacag aaaattagtt gcacggtgga tttggcccag 14880attttgcgtg agtatacatg gtcacacatc ttagagggta gaccattgat cggagcgaca 14940ttaccatgta tggtggagca attcaaagtt aagtggctaa gacaatatga accttgtcca 15000gaatgcctca acaaaaaagg ctcaaatgct tatgtctcag ttgcagtcaa agatcaagtg 15060gtcagtgctt ggcctaatac ttctcgaata agttggacaa tagggagtgg tgtcccctat 15120atagggtcaa gaaccgagga taaaatcgga cagcctgcaa tcaagccgcg atgcccttca 15180tctgccctca aggaggctat agaattagca tcaaggctca cttgggttac acaaggaagt 15240tctaatagtg aacaattaat ccggcctttc ttagaagcga gagtcaacct tagtgtcagt 15300gaagtcctgc aaatgacacc atcacattat tcaggaaata ttgtccatcg atataacgac 15360caatatagcc cgcactcatt tatggcgaat cgcatgagca atactgcgac ccgtctcata 15420gtgtcaacta atacacttgg agaattttca ggtggagggc aggccgccag ggatagcaat 15480ataattttcc agaatgttat aaatttagca gttgcccttt atgatattag attccggaat 15540acgaacacct ctgatataag gcataatagg gctcatcttc acctgacaga gtgctgtact 15600aaagaggtcc cggcccagta tttgacatat acaagtgcac tcaatctgga tttaagccgt 15660tatcgtgata atgaactaat atatgactca aatccactga ggggaggatt gaactgcaat 15720ttaacaatgg atagtccttt agtgaagggt cctaggctta acatgattga agatgatctt 15780ctccgctttc cacacctttc tggatgggag ttagcgaaaa cggtggtaca atccatcatc 15840tcagacaata gcaactcatc aacagatcca atcagtagcg gagaaacacg ctctttcaca 15900actcattttc tcacttaccc tcagattggc cttctttaca gtttcggggc agtattatgc 15960ttttatctag gcaatactat cctatggact aaaaaacttg attatgaaca gtttctatat 16020tatttgcata accagctgca caacttacct catcgagcac tccgtgtttt taaaccaaca 16080tttaagcatg ccagtgtgat gtcccgatta atggaaattg attccaactt ctcaatttat 16140attggcggga catctggaga tcgagggctg tctgatgctg ctcgactgtt tcttcggaca 16200gcaatcgcga gttttttaca atttcttaaa agctggatca tcgatcgcca aaaggcaatt 16260cctttatgga tagtatatcc gcttgaaggt caacagccgg aatccatcaa tgaatttcta 16320cataaaattt ttggtctgct caaacaaggc cccaaaaata ttccaaagga ggtcagcatt 16380caaaatgatg gacatttgga tttggcagaa aataattatg tttacaatag taagagcact 16440gctagtaatt tcttccatgc atccttagct tactggagaa gtaggaaatc tcggaaaact 16500caagaccata atgatttctc aagaggggat ggaacactta cagaacccgt gtgtaagttc 16560tcaagcaatc atcagtcaga tgaaaagtac tacaatgtga catgtggaaa gtcaccgaag 16620ccgcaagaac gcaaagactt ctcgcaatac agactcagca ataacgggca aacaatgagt 16680aatcatcgta agaaagggaa gttccacaag tggaatccct gcaaagtgtt aatggagagt 16740caaaggggaa ctgttctaaa agagggtgac tactttcaaa acaatactcc accaacagat 16800gatgtatcaa gtcctcaccg actcattcta ccatttttta aattgggaaa tcacaaccat 16860gcacatgatc aagatgccca agaattgata aatcaaaata ttaaacagta cctacatcag 16920ctaaggtcta tgttggacac cactatatat tgtagattca cagggattgt ctcatccatg 16980cattacaaat tggacgaagt tcttctagaa tacaatagtt tcgattcagc tatcacatta 17040gctgaaggtg aggggtcagg ggctctatta cttttgcaga aatatagtac aaggttatta 17100tttttgaaca cattggcaac agaacacagt atagagtcag aagttgtatc aggtttttct 17160actccgagaa tgttgttacc aataatgcaa aaggttcatg aaggacaagt cactgttatc 17220ttaaataatt cagcaagtca gataactgac ataactagct caatgtggtt aagtaatcaa 17280aaatataatc taccttgtca agttgaaatc attacgatgg atgctgaaac aacagagaac 17340ttaaacaggt cccaactcta ccgagcagta tataacttaa tacttgatca cattgatccg 17400cagtatctca aggtggtggt actcaaagta tttctgagtg atatagaagg aatattatgg 17460attaatgatt acttggctcc attattcggg gctggttact tgattaaacc gattacatca 17520agtgcccggt caagtgaatg gtacctttgc ttatcaaatt tgatatctac taacaggaga 17580tcggcccatc agactcacaa ggcatgtctt ggtgttatca gagatgcttt gcaagcacaa 17640gtccagcgag gcgtgtactg gttgagtcac atcgcacagt atgctacaaa gaatctccat 17700tgtgaataca tatgccttgg tttcccacct ctagaaaagg tcctatatca caggtataat 17760ctagttgata ctggactcgg tccattgtcg tcagttatta gacatttaac taacctccag 17820gcagagatac gagacttagt attagattat accctgatga gagagagtcg cactcaaacg 17880taccatttta ttaagactgc aaaaggcaga atcacaaagt tagtcaatga ctttctgaag 17940ttttctttaa ttgtccaggc actcaaaaat aattcttctt ggtatactga gcttaaaaaa 18000ttacctgagg tgattaatgt gtgtaatcga ttttatcata ctcacagttg cgaatgtcag 18060gaaaaattct ttgtccagac gctttattta caacgcctac gcgatgcaga aatcaagcta 18120attgaacgcc ttaccgggtt aatgcgattt tatccagaag ggttaatata ttccaatcac 18180acataggtac taaatcatca tagtatgagg aataaaataa tgataattcc tgacgacagt 18240tttagttccg attctaagta tatcggaaga gagtatgcca atcttaatta ttaaaggtaa 18300caagctatta gttattactt attgataaga ataaacttta tcatagcgta acacatcata 18360actttatagc gattttgcat ttctaatcct agtatttatt agaatgtact atcagagaaa 18420tgaccccagt tcctatcttt aaataatgat tgtgtgtatt aaattattag tttattaggt 18480ttatgagttg gttacacagt gagtattagt aattgaggat tatgtagata ggtaatctaa 18540cactgaatca cccatctgat gtcaccatat ccaaatattg tgctagtcgc atttaaacat 18600gctatcttca gttaagtaac atagactgaa aatgctaaga agagattgga gtaaaagtat 18660aaaataaatt taattaaact tcaaagtgat taaatgataa tgatcttggg aactcgatat 18720gacctcaagt caaaaataat gtcaatataa ttgtttagta atatgagtta taatgtgaat 18780tttgataact aactagcttt agtagttaag atcaaatgca aacattctaa gaatgttaag 18840cgcacacaaa aacattataa aaaaccaatt ttttcctttt tgtgtgtccc 1889022287PRTReston ebolavirus 22Met Glu His Ser Arg Glu Arg Gly Arg Ser Ser Asn Met Arg His Asn1 5 10 15 Ser Arg Glu Pro Tyr Glu Asn Pro Ser Arg Ser Arg Ser Leu Ser Arg 20 25 30 Asp Pro Asn Gln Val Asp Arg Arg Gln Pro Arg Ser Ala Ser Gln Ile 35 40 45 Arg Val Pro Asn Leu Phe His Arg Lys Lys Thr Asp Ala Leu Ile Val 50 55 60 Pro Pro Ala Pro Lys Asp Ile Cys Pro Thr Leu Lys Lys Gly Phe Leu65 70 75 80 Cys Asp Ser Lys Phe Cys Lys Lys Asp His Gln Leu Asp Ser Leu Asn 85 90 95 Asp His Glu Leu Leu Leu Leu Ile Ala Arg Arg Thr Cys Gly Ile Ile 100 105 110 Glu Ser Asn Ser Gln Ile Thr Ser Pro Lys Asp Met Arg Leu Ala Asn 115 120 125 Pro Thr Ala Glu Asp Phe Ser Gln Gly Asn Ser Pro Lys Leu Thr Leu 130 135 140 Ala Val Leu Leu Gln Ile Ala Glu His Trp Ala Thr Arg Asp Leu Arg145 150 155 160 Gln Ile Glu Asp Ser Lys Leu Arg Ala Leu Leu Thr Leu Cys Ala Val 165 170 175 Leu Thr Arg Lys Phe Ser Lys Ser Gln Leu Gly Leu Leu Cys Glu Thr 180 185 190 His Leu Arg His Glu Gly Leu Gly Gln Asp Gln Ala Asp Ser Val Leu 195 200 205 Glu Val Tyr Gln Arg Leu His Ser Asp Lys Gly Gly Asn Phe Glu Ala 210 215 220 Ala Leu Trp Gln Gln Trp Asp Arg Gln Ser Leu Ile Met Phe Ile Ser225 230 235 240 Ala Phe Leu Asn Ile Ala Leu Gln Thr Pro Cys Glu Ser Ser Ser Val 245 250 255 Val Val Ser Gly Leu Ala Thr Leu Tyr Pro Ala Gln Asp Asn Ser Thr 260 265 270 Pro Ser Glu Ala Thr Asn Asp Thr Thr Trp Ser Ser Thr Val Glu 275 280 285 2318959DNAZaire ebolavirus 23cggacacaca aaaagaaaga agaattttta ggatcttttg tgtgcgaata actatgagga 60agattaataa ttttcctctc attgaaattt atatcggaat ttaaattgaa attgttactg 120taatcacacc tggtttgttt cagagccaca tcacaaagat agagaacaac ctaggtctcc 180gaagggagca agggcatcag tgtgctcagt tgaaaatccc ttgtcaacac ctaggtctta 240tcacatcaca agttccacct cagactctgc agggtgatcc aacaacctta atagaaacat 300tattgttaaa ggacagcatt agttcacagt caaacaagca agattgagaa ttaaccttgg 360ttttgaactt gaacacttag gggattgaag attcaacaac cctaaagctt ggggtaaaac 420attggaaata gttaaaagac aaattgctcg gaatcacaaa attccgagta tggattctcg 480tcctcagaaa atctggatgg cgccgagtct cactgaatct gacatggatt accacaagat 540cttgacagca ggtctgtccg ttcaacaggg gattgttcgg caaagagtca tcccagtgta 600tcaagtaaac aatcttgaag aaatttgcca acttatcata caggcctttg aagcaggtgt 660tgattttcaa gagagtgcgg acagtttcct tctcatgctt tgtcttcatc atgcgtacca 720gggagattac aaacttttct tggaaagtgg cgcagtcaag tatttggaag ggcacgggtt 780ccgttttgaa gtcaagaagc gtgatggagt gaagcgcctt gaggaattgc tgccagcagt 840atctagtgga aaaaacatta agagaacact tgctgccatg ccggaagagg agacaactga 900agctaatgcc ggtcagtttc tctcctttgc aagtctattc cttccgaaat tggtagtagg 960agaaaaggct tgccttgaga aggttcaaag gcaaattcaa gtacatgcag agcaaggact 1020gatacaatat ccaacagctt ggcaatcagt aggacacatg atggtgattt tccgtttgat 1080gcgaacaaat tttctgatca aatttctcct aatacaccaa gggatgcaca tggttgccgg 1140gcatgatgcc aacgatgctg tgatttcaaa ttcagtggct caagctcgtt tttcaggctt 1200attgattgtc aaaacagtac ttgatcatat cctacaaaag acagaacgag gagttcgtct 1260ccatcctctt gcaaggaccg ccaaggtaaa aaatgaggtg aactccttta aggctgcact 1320cagctccctg gccaagcatg gagagtatgc tcctttcgcc cgacttttga acctttctgg 1380agtaaataat cttgagcatg gtcttttccc tcaactatcg gcaattgcac tcggagtcgc 1440cacagcacac gggagtaccc tcgcaggagt aaatgttgga gaacagtatc aacaactcag 1500agaggctgcc actgaggctg agaagcaact ccaacaatat gcagagtctc gcgaacttga 1560ccatcttgga cttgatgatc aggaaaagaa aattcttatg aacttccatc agaaaaagaa 1620cgaaatcagc ttccagcaaa caaacgctat ggtaactcta agaaaagagc gcctggccaa 1680gctgacagaa gctatcactg ctgcgtcact gcccaaaaca agtggacatt acgatgatga 1740tgacgacatt ccctttccag gacccatcaa tgatgacgac aatcctggcc atcaagatga 1800tgatccgact gactcacagg atacgaccat tcccgatgtg gtggttgatc ccgatgatgg 1860aagctacggc gaataccaga gttactcgga aaacggcatg aatgcaccag atgacttggt 1920cctattcgat ctagacgagg acgacgagga cactaagcca gtgcctaata gatcgaccaa 1980gggtggacaa cagaagaaca gtcaaaaggg ccagcatata gagggcagac agacacaatc 2040caggccaatt caaaatgtcc caggccctca cagaacaatc caccacgcca gtgcgccact 2100cacggacaat gacagaagaa atgaaccctc cggctcaacc agccctcgca tgctgacacc 2160aattaacgaa gaggcagacc cactggacga tgccgacgac gagacgtcta gccttccgcc 2220cttggagtca gatgatgaag agcaggacag ggacggaact tccaaccgca cacccactgt 2280cgccccaccg gctcccgtat acagagatca ctctgaaaag aaagaactcc cgcaagacga 2340gcaacaagat caggaccaca ctcaagaggc caggaaccag gacagtgaca acacccagtc 2400agaacactct tttgaggaga tgtatcgcca cattctaaga tcacaggggc catttgatgc 2460tgttttgtat tatcatatga tgaaggatga gcctgtagtt ttcagtacca gtgatggcaa 2520agagtacacg tatccagact cccttgaaga ggaatatcca ccatggctca ctgaaaaaga 2580ggctatgaat gaagagaata gatttgttac attggatggt caacaatttt attggccggt 2640gatgaatcac aagaataaat tcatggcaat cctgcaacat catcagtgaa tgagcatgga 2700acaatgggat gattcaaccg acaaatagct aacattaagt agtcaaggaa cgaaaacagg 2760aagaattttt gatgtctaag gtgtgaatta ttatcacaat aaaagtgatt cttatttttg 2820aatttaaagc tagcttatta ttactagccg tttttcaaag ttcaatttga gtcttaatgc 2880aaataggcgt taagccacag ttatagccat aattgtaact caatattcta actagcgatt 2940tatctaaatt aaattacatt atgcttttat aacttaccta ctagcctgcc caacatttac 3000acgatcgttt tataattaag aaaaaactaa tgatgaagat taaaaccttc atcatcctta 3060cgtcaattga attctctagc actcgaagct tattgtcttc aatgtaaaag aaaagctggt 3120ctaacaagat gacaactaga acaaagggca ggggccatac tgcggccacg actcaaaacg 3180acagaatgcc aggccctgag ctttcgggct ggatctctga gcagctaatg accggaagaa 3240ttcctgtaag cgacatcttc tgtgatattg agaacaatcc aggattatgc tacgcatccc 3300aaatgcaaca aacgaagcca aacccgaaga cgcgcaacag tcaaacccaa acggacccaa 3360tttgcaatca tagttttgag gaggtagtac aaacattggc ttcattggct actgttgtgc 3420aacaacaaac catcgcatca gaatcattag aacaacgcat tacgagtctt gagaatggtc 3480taaagccagt ttatgatatg gcaaaaacaa tctcctcatt gaacagggtt tgtgctgaga 3540tggttgcaaa atatgatctt ctggtgatga caaccggtcg ggcaacagca accgctgcgg 3600caactgaggc ttattgggcc gaacatggtc aaccaccacc tggaccatca

ctttatgaag 3660aaagtgcgat tcggggtaag attgaatcta gagatgagac cgtccctcaa agtgttaggg 3720aggcattcaa caatctaaac agtaccactt cactaactga ggaaaatttt gggaaacctg 3780acatttcggc aaaggatttg agaaacatta tgtatgatca cttgcctggt tttggaactg 3840ctttccacca attagtacaa gtgatttgta aattgggaaa agatagcaac tcattggaca 3900tcattcatgc tgagttccag gccagcctgg ctgaaggaga ctctcctcaa tgtgccctaa 3960ttcaaattac aaaaagagtt ccaatcttcc aagatgctgc tccacctgtc atccacatcc 4020gctctcgagg tgacattccc cgagcttgcc agaaaagctt gcgtccagtc ccaccatcgc 4080ccaagattga tcgaggttgg gtatgtgttt ttcagcttca agatggtaaa acacttggac 4140tcaaaatttg agccaatctc ccttccctcc gaaagaggcg aataatagca gaggcttcaa 4200ctgctgaact atagggtacg ttacattaat gatacacttg tgagtatcag ccctggataa 4260tataagtcaa ttaaacgacc aagataaaat tgttcatatc tcgctagcag cttaaaatat 4320aaatgtaata ggagctatat ctctgacagt attataatca attgttatta agtaacccaa 4380accaaaagtg atgaagatta agaaaaacct acctcggctg agagagtgtt ttttcattaa 4440ccttcatctt gtaaacgttg agcaaaattg ttaaaaatat gaggcgggtt atattgccta 4500ctgctcctcc tgaatatatg gaggccatat accctgtcag gtcaaattca acaattgcta 4560gaggtggcaa cagcaataca ggcttcctga caccggagtc agtcaatggg gacactccat 4620cgaatccact caggccaatt gccgatgaca ccatcgacca tgccagccac acaccaggca 4680gtgtgtcatc agcattcatc cttgaagcta tggtgaatgt catatcgggc cccaaagtgc 4740taatgaagca aattccaatt tggcttcctc taggtgtcgc tgatcaaaag acctacagct 4800ttgactcaac tacggccgcc atcatgcttg cttcatacac tatcacccat ttcggcaagg 4860caaccaatcc acttgtcaga gtcaatcggc tgggtcctgg aatcccggat catcccctca 4920ggctcctgcg aattggaaac caggctttcc tccaggagtt cgttcttccg ccagtccaac 4980taccccagta tttcaccttt gatttgacag cactcaaact gatcacccaa ccactgcctg 5040ctgcaacatg gaccgatgac actccaacag gatcaaatgg agcgttgcgt ccaggaattt 5100catttcatcc aaaacttcgc cccattcttt tacccaacaa aagtgggaag aaggggaaca 5160gtgccgatct aacatctccg gagaaaatcc aagcaataat gacttcactc caggacttta 5220agatcgttcc aattgatcca accaaaaata tcatgggaat cgaagtgcca gaaactctgg 5280tccacaagct gaccggtaag aaggtgactt ctaaaaatgg acaaccaatc atccctgttc 5340ttttgccaaa gtacattggg ttggacccgg tggctccagg agacctcacc atggtaatca 5400cacaggattg tgacacgtgt cattctcctg caagtcttcc agctgtgatt gagaagtaat 5460tgcaataatt gactcagatc cagttttata gaatcttctc agggatagtg ataacatcta 5520tttagtaatc cgtccattag aggagacact tttaattgat caatatacta aaggtgcttt 5580acaccattgt cttttttctc tcctaaatgt agaacttaac aaaagactca taatatactt 5640gtttttaaag gattgattga tgaaagatca taactaataa cattacaaat aatcctacta 5700taatcaatac ggtgattcaa atgttaatct ttctcattgc acatactttt tgcccttatc 5760ctcaaattgc ctgcatgctt acatctgagg atagccagtg tgacttggat tggaaatgtg 5820gagaaaaaat cgggacccat ttctaggttg ttcacaatcc aagtacagac attgcccttc 5880taattaagaa aaaatcggcg atgaagatta agccgacagt gagcgtaatc ttcatctctc 5940ttagattatt tgttttccag agtaggggtc gtcaggtcct tttcaatcgt gtaaccaaaa 6000taaactccac tagaaggata ttgtggggca acaacacaat gggcgttaca ggaatattgc 6060agttacctcg tgatcgattc aagaggacat cattctttct ttgggtaatt atccttttcc 6120aaagaacatt ttccatccca cttggagtca tccacaatag cacattacag gttagtgatg 6180tcgacaaact agtttgtcgt gacaaactgt catccacaaa tcaattgaga tcagttggac 6240tgaatctcga agggaatgga gtggcaactg acgtgccatc tgcaactaaa agatggggct 6300tcaggtccgg tgtcccacca aaggtggtca attatgaagc tggtgaatgg gctgaaaact 6360gctacaatct tgaaatcaaa aaacctgacg ggagtgagtg tctaccagca gcgccagacg 6420ggattcgggg cttcccccgg tgccggtatg tgcacaaagt atcaggaacg ggaccgtgtg 6480ccggagactt tgccttccat aaagagggtg ctttcttcct gtatgatcga cttgcttcca 6540cagttatcta ccgaggaacg actttcgctg aaggtgtcgt tgcatttctg atactgcccc 6600aagctaagaa ggacttcttc agctcacacc ccttgagaga gccggtcaat gcaacggagg 6660acccgtctag tggctactat tctaccacaa ttagatatca ggctaccggt tttggaacca 6720atgagacaga gtacttgttc gaggttgaca atttgaccta cgtccaactt gaatcaagat 6780tcacaccaca gtttctgctc cagctgaatg agacaatata tacaagtggg aaaaggagca 6840ataccacggg aaaactaatt tggaaggtca accccgaaat tgatacaaca atcggggagt 6900gggccttctg ggaaactaaa aaaacctcac tagaaaaatt cgcagtgaag agttgtcttt 6960cacagttgta tcaaacggag ccaaaaacat cagtggtcag agtccggcgc gaacttcttc 7020cgacccaggg accaacacaa caactgaaga ccacaaaatc atggcttcag aaaattcctc 7080tgcaatggtt caagtgcaca gtcaaggaag ggaagctgca gtgtcgcatc taacaaccct 7140tgccacaatc tccacgagtc cccaatccct cacaaccaaa ccaggtccgg acaacagcac 7200ccataataca cccgtgtata aacttgacat ctctgaggca actcaagttg aacaacatca 7260ccgcagaaca gacaacgaca gcacagcctc cgacactccc tctgccacga ccgcagccgg 7320acccccaaaa gcagagaaca ccaacacgag caagagcact gacttcctgg accccgccac 7380cacaacaagt ccccaaaacc acagcgagac cgctggcaac aacaacactc atcaccaaga 7440taccggagaa gagagtgcca gcagcgggaa gctaggctta attaccaata ctattgctgg 7500agtcgcagga ctgatcacag gcgggagaag aactcgaaga gaagcaattg tcaatgctca 7560acccaaatgc aaccctaatt tacattactg gactactcag gatgaaggtg ctgcaatcgg 7620actggcctgg ataccatatt tcgggccagc agccgaggga atttacatag aggggctaat 7680gcacaatcaa gatggtttaa tctgtgggtt gagacagctg gccaacgaga cgactcaagc 7740tcttcaactg ttcctgagag ccacaactga gctacgcacc ttttcaatcc tcaaccgtaa 7800ggcaattgat ttcttgctgc agcgatgggg cggcacatgc cacattctgg gaccggactg 7860ctgtatcgaa ccacatgatt ggaccaagaa cataacagac aaaattgatc agattattca 7920tgattttgtt gataaaaccc ttccggacca gggggacaat gacaattggt ggacaggatg 7980gagacaatgg ataccggcag gtattggagt tacaggcgtt ataattgcag ttatcgcttt 8040attctgtata tgcaaatttg tcttttagtt tttcttcaga ttgcttcatg gaaaagctca 8100gcctcaaatc aatgaaacca ggatttaatt atatggatta cttgaatcta agattacttg 8160acaaatgata atataataca ctggagcttt aaacatagcc aatgtgattc taactccttt 8220aaactcacag ttaatcataa acaaggtttg acatcaatct agttatctct ttgagaatga 8280taaacttgat gaagattaag aaaaaggtaa tctttcgatt atctttaatc ttcatccttg 8340attctacaat catgacagtt gtctttagtg acaagggaaa gaagcctttt tattaagttg 8400taataatcag atctgcgaac cggtagagtt tagttgcaac ctaacacaca taaagcattg 8460gtcaaaaagt caatagaaat ttaaacagtg agtggagaca acttttaaat ggaagcttca 8520tatgagagag gacgcccacg agctgccaga cagcattcaa gggatggaca cgaccaccat 8580gttcgagcac gatcatcatc cagagagaat tatcgaggtg agtaccgtca atcaaggagc 8640gcctcacaag tgcgcgttcc tactgtattt cataagaaga gagttgaacc attaacagtt 8700cctccagcac ctaaagacat atgtccgacc ttgaaaaaag gatttttgtg tgacagtagt 8760ttttgcaaaa aagatcacca gttggagagt ttaactgata gggaattact cctactaatc 8820gcccgtaaga cttgtggatc agtagaacaa caattaaata taactgcacc caaggactcg 8880cgcttagcaa atccaacggc tgatgatttc cagcaagagg aaggtccaaa aattaccttg 8940ttgacactga tcaagacggc agaacactgg gcgagacaag acatcagaac catagaggat 9000tcaaaattaa gagcattgtt gactctatgt gctgtgatga cgaggaaatt ctcaaaatcc 9060cagctgagtc ttttatgtga gacacaccta aggcgcgagg ggcttgggca agatcaggca 9120gaacccgttc tcgaagtata tcaacgatta cacagtgata aaggaggcag ttttgaagct 9180gcactatggc aacaatggga ccgacaatcc ctaattatgt ttatcactgc attcttgaat 9240attgctctcc agttaccgtg tgaaagttct gctgtcgttg tttcagggtt aagaacattg 9300gttcctcaat cagataatga ggaagcttca accaacccgg ggacatgctc atggtctgat 9360gagggtaccc cttaataagg ctgactaaaa cactatataa ccttctactt gatcacaata 9420ctccgtatac ctatcatcat atatttaatc aagacgatat cctttaaaac ttattcagta 9480ctataatcac tctcgtttca aattaataag atgtgcatga ttgccctaat atatgaagag 9540gtatgataca accctaacag tgatcaaaga aaatcataat ctcgtatcgc tcgtaatata 9600acctgccaag catacctctt gcacaaagtg attcttgtac acaaataatg ttttactcta 9660caggaggtag caacgatcca tcccatcaaa aaataagtat ttcatgactt actaatgatc 9720tcttaaaata ttaagaaaaa ctgacggaac ataaattctt tatgcttcaa gctgtggagg 9780aggtgtttgg tattggctat tgttatatta caatcaataa caagcttgta aaaatattgt 9840tcttgtttca agaggtagat tgtgaccgga aatgctaaac taatgatgaa gattaatgcg 9900gaggtctgat aagaataaac cttattattc agattaggcc ccaagaggca ttcttcatct 9960ccttttagca aagtactatt tcagggtagt ccaattagtg gcacgtcttt tagctgtata 10020tcagtcgccc ctgagatacg ccacaaaagt gtctctaagc taaattggtc tgtacacatc 10080ccatacattg tattaggggc aataatatct aattgaactt agccgtttaa aatttagtgc 10140ataaatctgg gctaacacca ccaggtcaac tccattggct gaaaagaagc ttacctacaa 10200cgaacatcac tttgagcgcc ctcacaatta aaaaatagga acgtcgttcc aacaatcgag 10260cgcaaggttt caaggttgaa ctgagagtgt ctagacaaca aaatattgat actccagaca 10320ccaagcaaga cctgagaaaa aaccatggct aaagctacgg gacgatacaa tctaatatcg 10380cccaaaaagg acctggagaa aggggttgtc ttaagcgacc tctgtaactt cttagttagc 10440caaactattc aggggtggaa ggtttattgg gctggtattg agtttgatgt gactcacaaa 10500ggaatggccc tattgcatag actgaaaact aatgactttg cccctgcatg gtcaatgaca 10560aggaatctct ttcctcattt atttcaaaat ccgaattcca caattgaatc accgctgtgg 10620gcattgagag tcatccttgc agcagggata caggaccagc tgattgacca gtctttgatt 10680gaacccttag caggagccct tggtctgatc tctgattggc tgctaacaac caacactaac 10740catttcaaca tgcgaacaca acgtgtcaag gaacaattga gcctaaaaat gctgtcgttg 10800attcgatcca atattctcaa gtttattaac aaattggatg ctctacatgt cgtgaactac 10860aacggattgt tgagcagtat tgaaattgga actcaaaatc atacaatcat cataactcga 10920actaacatgg gttttctggt ggagctccaa gaacccgaca aatcggcaat gaaccgcatg 10980aagcctgggc cggcgaaatt ttccctcctt catgagtcca cactgaaagc atttacacaa 11040ggatcctcga cacgaatgca aagtttgatt cttgaattta atagctctct tgctatctaa 11100ctaaggtaga atacttcata ttgagctaac tcatatatgc tgactcaata gttatcttga 11160catctctgct ttcataatca gatatataag cataataaat aaatactcat atttcttgat 11220aatttgttta accacagata aatcctcact gtaagccagc ttccaagttg acacccttac 11280aaaaaccagg actcagaatc cctcaaacaa gagattccaa gacaacatca tagaattgct 11340ttattatatg aataagcatt ttatcaccag aaatcctata tactaaatgg ttaattgtaa 11400ctgaacccgc aggtcacatg tgttaggttt cacagattct atatattact aactctatac 11460tcgtaattaa cattagataa gtagattaag aaaaaagcct gaggaagatt aagaaaaact 11520gcttattggg tctttccgtg ttttagatga agcagttgaa attcttcctc ttgatattaa 11580atggctacac aacataccca atacccagac gctaggttat catcaccaat tgtattggac 11640caatgtgacc tagtcactag agcttgcggg ttatattcat catactccct taatccgcaa 11700ctacgcaact gtaaactccc gaaacatatc taccgtttga aatacgatgt aactgttacc 11760aagttcttga gtgatgtacc agtggcgaca ttgcccatag atttcatagt cccagttctt 11820ctcaaggcac tgtcaggcaa tggattctgt cctgttgagc cgcggtgcca acagttctta 11880gatgaaatca ttaagtacac aatgcaagat gctctcttct tgaaatatta tctcaaaaat 11940gtgggtgctc aagaagactg tgttgatgaa cactttcaag agaaaatctt atcttcaatt 12000cagggcaatg aatttttaca tcaaatgttt ttctggtatg atctggctat tttaactcga 12060aggggtagat taaatcgagg aaactctaga tcaacatggt ttgttcatga tgatttaata 12120gacatcttag gctatgggga ctatgttttt tggaagatcc caatttcaat gttaccactg 12180aacacacaag gaatccccca tgctgctatg gactggtatc aggcatcagt attcaaagaa 12240gcggttcaag ggcatacaca cattgtttct gtttctactg ccgacgtctt gataatgtgc 12300aaagatttaa ttacatgtcg attcaacaca actctaatct caaaaatagc agagattgag 12360gatccagttt gttctgatta tcccaatttt aagattgtgt ctatgcttta ccagagcgga 12420gattacttac tctccatatt agggtctgat gggtataaaa ttattaagtt cctcgaacca 12480ttgtgcttgg ccaaaattca attatgctca aagtacactg agaggaaggg ccgattctta 12540acacaaatgc atttagctgt aaatcacacc ctagaagaaa ttacagaaat gcgtgcacta 12600aagccttcac aggctcaaaa gatccgtgaa ttccatagaa cattgataag gctggagatg 12660acgccacaac aactttgtga gctattttcc attcaaaaac actgggggca tcctgtgcta 12720catagtgaaa cagcaatcca aaaagttaaa aaacatgcta cggtgctaaa agcattacgc 12780cctatagtga ttttcgagac atactgtgtt tttaaatata gtattgccaa acattatttt 12840gatagtcaag gatcttggta cagtgttact tcagatagga atctaacacc gggtcttaat 12900tcttatatca aaagaaatca attccctccg ttgccaatga ttaaagaact actatgggaa 12960ttttaccacc ttgaccaccc tccacttttc tcaaccaaaa ttattagtga cttaagtatt 13020tttataaaag acagagctac cgcagtagaa aggacatgct gggatgcagt attcgagcct 13080aatgttctag gatataatcc acctcacaaa tttagtacta aacgtgtacc ggaacaattt 13140ttagagcaag aaaacttttc tattgagaat gttctttcct acgcacaaaa actcgagtat 13200ctactaccac aatatcggaa cttttctttc tcattgaaag agaaagagtt gaatgtaggt 13260agaaccttcg gaaaattgcc ttatccgact cgcaatgttc aaacactttg tgaagctctg 13320ttagctgatg gtcttgctaa agcatttcct agcaatatga tggtagttac ggaacgtgag 13380caaaaagaaa gcttattgca tcaagcatca tggcaccaca caagtgatga ttttggtgaa 13440catgccacag ttagagggag tagctttgta actgatttag agaaatacaa tcttgcattt 13500agatatgagt ttacagcacc ttttatagaa tattgcaacc gttgctatgg tgttaagaat 13560gtttttaatt ggatgcatta tacaatccca cagtgttata tgcatgtcag tgattattat 13620aatccaccac ataacctcac actggagaat cgagacaacc cccccgaagg gcctagttca 13680tacaggggtc atatgggagg gattgaagga ctgcaacaaa aactctggac aagtatttca 13740tgtgctcaaa tttctttagt tgaaattaag actggtttta agttacgctc agctgtgatg 13800ggtgacaatc agtgcattac tgttttatca gtcttcccct tagagactga cgcagacgag 13860caggaacaga gcgccgaaga caatgcagcg agggtggccg ccagcctagc aaaagttaca 13920agtgcctgtg gaatcttttt aaaacctgat gaaacatttg tacattcagg ttttatctat 13980tttggaaaaa aacaatattt gaatggggtc caattgcctc agtcccttaa aacggctaca 14040agaatggcac cattgtctga tgcaattttt gatgatcttc aagggaccct ggctagtata 14100ggcactgctt ttgagcgatc catctctgag acacgacata tctttccttg caggataacc 14160gcagctttcc atacgttttt ttcggtgaga atcttgcaat atcatcatct cgggttcaat 14220aaaggttttg accttggaca gttaacactc ggcaaacctc tggatttcgg aacaatatca 14280ttggcactag cggtaccgca ggtgcttgga gggttatcct tcttgaatcc tgagaaatgt 14340ttctaccgga atctaggaga tccagttacc tcaggcttat tccagttaaa aacttatctc 14400cgaatgattg agatggatga tttattctta cctttaattg cgaagaaccc tgggaactgc 14460actgccattg actttgtgct aaatcctagc ggattaaatg tccctgggtc gcaagactta 14520acttcatttc tgcgccagat tgtacgcagg accatcaccc taagtgcgaa aaacaaactt 14580attaatacct tatttcatgc gtcagctgac ttcgaagacg aaatggtttg taaatggcta 14640ttatcatcaa ctcctgttat gagtcgtttt gcggccgata tcttttcacg cacgccgagc 14700gggaagcgat tgcaaattct aggatacctg gaaggaacac gcacattatt agcctctaag 14760atcatcaaca ataatacaga gacaccggtt ttggacagac tgaggaaaat aacattgcaa 14820aggtggagcc tatggtttag ttatcttgat cattgtgata atatcctggc ggaggcttta 14880acccaaataa cttgcacagt tgatttagca cagattctga gggaatattc atgggctcat 14940attttagagg gaagacctct tattggagcc acactcccat gtatgattga gcaattcaaa 15000gtgttttggc tgaaacccta cgaacaatgt ccgcagtgtt caaatgcaaa gcaaccaggt 15060gggaaaccat tcgtgtcagt ggcagtcaag aaacatattg ttagtgcatg gccgaacgca 15120tcccgaataa gctggactat cggggatgga atcccataca ttggatcaag gacagaagat 15180aagataggac aacctgctat taaaccaaaa tgtccttccg cagccttaag agaggccatt 15240gaattggcgt cccgtttaac atgggtaact caaggcagtt cgaacagtga cttgctaata 15300aaaccatttt tggaagcacg agtaaattta agtgttcaag aaatacttca aatgacccct 15360tcacattact caggaaatat tgttcacagg tacaacgatc aatacagtcc tcattctttc 15420atggccaatc gtatgagtaa ttcagcaacg cgattgattg tttctacaaa cactttaggt 15480gagttttcag gaggtggcca gtctgcacgc gacagcaata ttattttcca gaatgttata 15540aattatgcag ttgcactgtt cgatattaaa tttagaaaca ctgaggctac agatatccaa 15600tataatcgtg ctcaccttca tctaactaag tgttgcaccc gggaagtacc agctcagtat 15660ttaacataca catctacatt ggatttagat ttaacaagat accgagaaaa cgaattgatt 15720tatgacagta atcctctaaa aggaggactc aattgcaata tctcattcga taatccattt 15780ttccaaggta aacggctgaa cattatagaa gatgatctta ttcgactgcc tcacttatct 15840ggatgggagc tagccaagac catcatgcaa tcaattattt cagatagcaa caattcatct 15900acagacccaa ttagcagtgg agaaacaaga tcattcacta cccatttctt aacttatccc 15960aagataggac ttctgtacag ttttggggcc tttgtaagtt attatcttgg caatacaatt 16020cttcggacta agaaattaac acttgacaat tttttatatt acttaactac tcaaattcat 16080aatctaccac atcgctcatt gcgaatactt aagccaacat tcaaacatgc aagcgttatg 16140tcacggttaa tgagtattga tcctcatttt tctatttaca taggcggtgc tgcaggtgac 16200agaggactct cagatgcggc caggttattt ttgagaacgt ccatttcatc ttttcttaca 16260tttgtaaaag aatggataat taatcgcgga acaattgtcc ctttatggat agtatatccg 16320ctagagggtc aaaacccaac acctgtgaat aattttctct atcagatcgt agaactgctg 16380gtgcatgatt catcaagaca acaggctttt aaaactacca taagtgatca tgtacatcct 16440cacgacaatc ttgtttacac atgtaagagt acagccagca atttcttcca tgcatcattg 16500gcgtactgga ggagcagaca cagaaacagc aaccgaaaat acttggcaag agactcttca 16560actggatcaa gcacaaacaa cagtgatggt catattgaga gaagtcaaga acaaaccacc 16620agagatccac atgatggcac tgaacggaat ctagtcctac aaatgagcca tgaaataaaa 16680agaacgacaa ttccacaaga aaacacgcac cagggtccgt cgttccagtc ctttctaagt 16740gactctgctt gtggtacagc aaatccaaaa ctaaatttcg atcgatcgag acacaatgtg 16800aaatttcagg atcataactc ggcatccaag agggaaggtc atcaaataat ctcacaccgt 16860ctagtcctac ctttctttac attatctcaa gggacacgcc aattaacgtc atccaatgag 16920tcacaaaccc aagacgagat atcaaagtac ttacggcaat tgagatccgt cattgatacc 16980acagtttatt gtagatttac cggtatagtc tcgtccatgc attacaaact tgatgaggtc 17040ctttgggaaa tagagagttt caagtcggct gtgacgctag cagagggaga aggtgctggt 17100gccttactat tgattcagaa ataccaagtt aagaccttat ttttcaacac gctagctact 17160gagtccagta tagagtcaga aatagtatca ggaatgacta ctcctaggat gcttctacct 17220gttatgtcaa aattccataa tgaccaaatt gagattattc ttaacaactc agcaagccaa 17280ataacagaca taacaaatcc tacttggttt aaagaccaaa gagcaaggct acctaagcaa 17340gtcgaggtta taaccatgga tgcagagaca acagagaata taaacagatc gaaattgtac 17400gaagctgtat ataaattgat cttacaccat attgatccta gcgtattgaa agcagtggtc 17460cttaaagtct ttctaagtga tactgagggt atgttatggc taaatgataa tttagccccg 17520ttttttgcca ctggttattt aattaagcca ataacgtcaa gtgctagatc tagtgagtgg 17580tatctttgtc tgacgaactt cttatcaact acacgtaaga tgccacacca aaaccatctc 17640agttgtaaac aggtaatact tacggcattg caactgcaaa ttcaacgaag cccatactgg 17700ctaagtcatt taactcagta tgctgactgt gagttacatt taagttatat ccgccttggt 17760tttccatcat tagagaaagt actataccac aggtataacc tcgtcgattc aaaaagaggt 17820ccactagtct ctatcactca gcacttagca catcttagag cagagattcg agaattaact 17880aatgattata atcaacagcg acaaagtcgg actcaaacat atcactttat tcgtactgca 17940aaaggacgaa tcacaaaact agtcaatgat tatttaaaat tctttcttat tgtgcaagca 18000ttaaaacata atgggacatg gcaagctgag tttaagaaat taccagagtt gattagtgtg 18060tgcaataggt tctaccatat tagagattgc aattgtgaag aacgtttctt agttcaaacc 18120ttatatttac atagaatgca ggattctgaa gttaagctta tcgaaaggct gacagggctt 18180ctgagtttat ttccggatgg tctctacagg tttgattgaa ttaccgtgca tagtatcctg 18240atacttgcaa aggttggtta ttaacataca gattataaaa aactcataaa ttgctctcat 18300acatcatatt gatctaatct caataaacaa ctatttaaat aacgaaagga gtccctatat 18360tatatactat atttagcctc tctccctgcg tgataatcaa aaaattcaca atgcagcatg 18420tgtgacatat tactgccgca atgaatttaa cgcaacataa taaactctgc actctttata 18480attaagcttt aacgaaaggt ctgggctcat attgttattg atataataat gttgtatcaa 18540tatcctgtca gatggaatag tgttttggtt gataacacaa cttcttaaaa caaaattgat 18600ctttaagatt aagtttttta taattatcat tactttaatt tgtcgtttta aaaacggtga 18660tagccttaat ctttgtgtaa aataagagat taggtgtaat aaccttaaca

tttttgtcta 18720gtaagctact atttcataca gaatgataaa attaaaagaa aaggcaggac tgtaaaatca 18780gaaatacctt ctttacaata tagcagacta gataataatc ttcgtgttaa tgataattaa 18840gacattgacc acgctcatca gaaggctcgc cagaataaac gttgcaaaaa ggattcctgg 18900aaaaatggtc gcacacaaaa atttaaaaat aaatctattt cttctttttt gtgtgtcca 1895924288PRTZaire ebolavirus 24Met Glu Ala Ser Tyr Glu Arg Gly Arg Pro Arg Ala Ala Arg Gln His1 5 10 15 Ser Arg Asp Gly His Asp His His Val Arg Ala Arg Ser Ser Ser Arg 20 25 30 Glu Asn Tyr Arg Gly Glu Tyr Arg Gln Ser Arg Ser Ala Ser Gln Val 35 40 45 Arg Val Pro Thr Val Phe His Lys Lys Arg Val Glu Pro Leu Thr Val 50 55 60 Pro Pro Ala Pro Lys Asp Ile Cys Pro Thr Leu Lys Lys Gly Phe Leu65 70 75 80 Cys Asp Ser Ser Phe Cys Lys Lys Asp His Gln Leu Glu Ser Leu Thr 85 90 95 Asp Arg Glu Leu Leu Leu Leu Ile Ala Arg Lys Thr Cys Gly Ser Val 100 105 110 Glu Gln Gln Leu Asn Ile Thr Ala Pro Lys Asp Ser Arg Leu Ala Asn 115 120 125 Pro Thr Ala Asp Asp Phe Gln Gln Glu Glu Gly Pro Lys Ile Thr Leu 130 135 140 Leu Thr Leu Ile Lys Thr Ala Glu His Trp Ala Arg Gln Asp Ile Arg145 150 155 160 Thr Ile Glu Asp Ser Lys Leu Arg Ala Leu Leu Thr Leu Cys Ala Val 165 170 175 Met Thr Arg Lys Phe Ser Lys Ser Gln Leu Ser Leu Leu Cys Glu Thr 180 185 190 His Leu Arg Arg Glu Gly Leu Gly Gln Asp Gln Ala Glu Pro Val Leu 195 200 205 Glu Val Tyr Gln Arg Leu His Ser Asp Lys Gly Gly Ser Phe Glu Ala 210 215 220 Ala Leu Trp Gln Gln Trp Asp Arg Gln Ser Leu Ile Met Phe Ile Thr225 230 235 240 Ala Phe Leu Asn Ile Ala Leu Gln Leu Pro Cys Glu Ser Ser Ala Val 245 250 255 Val Val Ser Gly Leu Arg Thr Leu Val Pro Gln Ser Asp Asn Glu Glu 260 265 270 Ala Ser Thr Asn Pro Gly Thr Cys Ser Trp Ser Asp Glu Gly Thr Pro 275 280 285 2519111DNALake Victoria ebolavirus 25agacacacaa aaacaagaga tgatgatttt gtgtatcata taaataaaga agaatattaa 60cattgacatt gagacttgtc agtctgttaa tattcttgaa gagatggatt tacacagttt 120gttggagttg ggtacaaaac ccactgcccc tcatgttcgt aataagaaag tgatattatt 180tgacacaaat catcaggtta gtatctgtaa tcagataata gatgcaataa actcagggat 240tgatcttgga gatctcctag aagggggttt gctgacgttg tgtgttgagc attactataa 300ttctgataag gataaattca acacaagtcc tatcgcgaag tacttacgtg atgcgggcta 360tgaatttgat gtcatcaaga atgcagatgc aacccgcttt ctggatgtga ttcctaatga 420acctcattac agccctttaa ttctagccct taagacattg gaaagtactg aatctcagag 480ggggagaatt gggctctttt tatcattttg cagtcttttc ctcccaaaac ttgtcgtcgg 540agaccgagct agtatcgaaa aggctttaag acaagtaaca gtgcatcaag aacaggggat 600cgtcacatac cctaatcatt ggcttaccac aggccacatg aaagtaattt tcgggatttt 660gaggtccagc ttcattttaa agtttgtgtt gattcatcaa ggagtaaatt tggtgacagg 720tcatgatgcc tatgacagta tcattagtaa ttcagtaggt caaactagat tctcaggact 780tcttatcgtg aaaacagttc tcgagttcat cttgcaaaaa actgattcag gggtgacact 840acatcctttg gtgcggacct ccaaagtaaa aaatgaagtt gctagtttca agcaggcgtt 900gagcaaccta gcccgacatg gggaatacgc accatttgca cgggttctga atttatcagg 960gattaacaac ctcgaacatg gactctatcc tcagctttca gcaattgcgc tgggtgtggc 1020aacagcacac ggcagtacat tggctggtgt caatgttggc gaacaatatc aacaactacg 1080agaggcggca catgatgcgg aagtaaaact acaaaggcga catgaacatc aggaaattca 1140agctattgcc gaggatgacg aggaaaggaa gatattagaa caattccacc ttcagaaaac 1200tgaaatcaca cacagtcaga cactagccgt cctcagccag aaacgagaaa aattagctcg 1260tctcgctgca gaaattgaaa acaatattgt ggaagatcag ggatttaagc aatcacagaa 1320tcgggtgtca cagtcgtttt tgaatgaccc tacacctgtg gaagtaacgg ttcaagccag 1380gcccatgaat cgaccaactg ctctgcctcc cccagttgac gacaagattg agcatgaatc 1440tacagaagat agctcttctt caagtagctt tgttgacttg aatgatccat ttgcactgct 1500gaatgaggac gaggatactc ttgatgacag tgtcatgatc ccgggcacaa catcgagaga 1560atttcaaggg attcctgaac cgccaagaca atcccaagac ctcaataaca gccaaggaaa 1620gcaggaagat gaatccacaa atccgattaa gaaacagttt ctgagatatc aagaattgcc 1680tcctgttcaa gaggatgatg aatcggaata cacaactgac tctcaagaaa gcatcgacca 1740accaggatcc gacaatgaac aaggagttga tcttccacct cctccgttgt acgctcagga 1800aaaaagacag gacccaatac agcacccagc agcaaaccct caggatccct tcggcagtat 1860tggtgatgta aatggtgata tcttagaacc tataagatca ccttcttcac catctgctcc 1920tcaggaagac acaaggatga gggaagccta tgaattgtcg cctgatttca caaatgatga 1980ggataatcag cagaattggc cacaaagagt ggtgacaaag aagggtagaa ctttccttta 2040tcctaatgat cttctgcaaa caaatcctcc agagtcactt ataacagccc tcgttgagga 2100ataccaaaat cctgtctcag ctaaggagct tcaagcagat tggcccgaca tgtcatttga 2160tgaaaggaga catgttgcga tgaacttgta gtccagataa cacagcacgg ttactcactt 2220atctattttg atatgactca tcctcagatc acagcaatca aatttatttg aatatttgaa 2280ccacctttta gtatcctatt acttgttact attgtgtgag acaacataag ccatcaaata 2340acaatcacgg gcaaggactg ggcatactat ggtggtctta gagcattgtc cagtgctaca 2400aattcttttt tcaattgcta taattataca actacaaacc tccatacatt tgccgcaaca 2460ctgtaatcaa cactgctgta tctcttcttc aagccatctg atttaactta ataaacatga 2520cttgattcaa agaatatact gacaatgtta ctgtttgaat ttctcaagtg gtgcactatc 2580ctactgtttt gctcagctta gtatattgta atatgtaagt ggactctccc cttctcctct 2640cgtgtattct ttataaatca cttacttgat agaatgtcga gtctactggt ttggagtttc 2700cttactctaa tggatgtaat aattaactgt tggcctagat gataacagat atgaggttat 2760ataattactc atagtgtaaa gtataattct tacctctgtt tcttctgttt tccctttctt 2820ttataatatg ccaattaaga aaaactaaaa atcgaagaat attaaagatt ttctttaata 2880ttcagaaaag gctttttatt ctattctttc tttttacaaa cgtattgaaa tagtaattct 2940cacaatgtgg gactcatcat acatgcagca agtcagcgaa gggttgatga ctggaaaagt 3000acccatagat caagtgtttg gtgccaatcc cttagagaag ttatacaaga gaagaaaacc 3060aaaaggcaca gttggactac aatgtagccc ttgtctaatg tcaaaggcga caagtactga 3120tgatattatt tgggaccaac tgatcgtgaa gagaacacta gctgatctac ttataccgat 3180aaataggcag atatcagaca ttcaaagcac tctaagcgaa gtaacaacaa gagtccatga 3240aattgagcgg caattacatg agattacccc agttttaaaa atgggaagga cactggaagc 3300aatttccaag gggatgtcag aaatgttagc caaatacgac caccttgtaa tttcaactgg 3360aagaaccact gcaccagctg ctgcctttga tgcctactta aatgagcatg gtgtccctcc 3420ccctcaaccc gcgattttca aagatcttgg ggttgcccaa caagcttgta gtaaggggac 3480catggttaaa aatgcaacaa cagatgcagc cgacaagatg tcgaaggttc ttgaactcag 3540tgaggaaacg ttctccaagc caaacctttc agctaaggat ttagcccttt tattgtttac 3600ccatctaccc ggcaacaaca ctccattcca tatcctagct caggtccttt caaaaattgc 3660ttacaagtca ggaaaatccg gagcattctt ggatgcattt caccagattc taagtgaagg 3720agagaatgct caggcggcat taactcgact aagcagaaca tttgacgctt tccttggagt 3780ggttcctcca gtgataagag tcaaaaactt ccaaacagtc cctcgtccat gtcaaaaaag 3840tcttcgggct gtccctccaa atccaacaat tgacaaagga tgggtctgtg tttattcatc 3900tgagcaaggt gaaacacggg cccttaaaat ctaattctca ttgttcatag ttgcaaggga 3960agtgatcttt ccgagttgat acaaagacac taaacatttc aaaagcatgt atgtggacaa 4020aacataatta gaccatctta attggagtag taatttattt ctgtcttaaa tgtgattttc 4080actttaaaag cgttaaatgg tgatagatta atccttgaag ttactcttct atatattata 4140gagaaaccaa tgttactaac aaaaggggtc tacctaacgc atatgattga gtaatccgta 4200tattttataa accaaacaat taacttctta ctttttaaga atcaactaac aacatagaaa 4260agacatttat ccttatgtaa tcctcggctt agttgaaatt aacttttgtt ggacctcaag 4320acgcttattc atagtatatt atatgatttt ttataagttt aagatatctt aaattatacc 4380cacaaaagat actgttttaa ttaagaaaaa ctatgaagaa cattaagaag atctttcttt 4440cgtagtgttc ttttactgga aggagtattc caatttcagc ttgttggatt aattgttact 4500taaattgtcc tttttgaaat taattcacac aaggtagttt aaatttatat ccaaaataaa 4560ttttgatatg gccagttcca gcaattacaa cacatacatg caatacttga acccccctcc 4620ttatgctgat cacggtgcaa accagttgat cccggcggat cagctatcaa atcagcaggg 4680tataactcca aattacgtgg gtgatttaaa cctagatgat cagttcaaag ggaatgtctg 4740ccatgctttc actttagagg caataattga catatctgca tataacgagc gaacagtcaa 4800aggcgttccg gcatggctgc ctcttgggat tatgagcaat tttgaatatc ctttagctca 4860tactgtggcc gcgttgctca caggcagcta tacaatcacc caatttactc acaacgggca 4920aaaattcgtc cgtgttaatc gacttggtac aggaatccca gcacacccac tcagaatgtt 4980gcgtgaagga aatcaagctt ttattcagaa tatggtgatc cccaggaatt tttcaactaa 5040tcaattcacc tacaatctca ctaatttagt attgagtgtg caaaaacttc ctgatgatgc 5100ctggcgccca tccaaggaca aattaattgg gaacactatg catcccgcag tctccatcca 5160cccgaatctg ccgcctattg ttctaccaac agtcaagaag caggcttatc gtcagcacaa 5220aaatcccaac aatggaccat tgctggccat atctggcatc ctccatcaac tgagggtcga 5280aaaagtccca gagaagacga gcctgtttag gatctcgctt cctgccgaca tgttctcagt 5340aaaagagggt atgatgaaga aaaggggaga aaattccccc gtggtttatt ttcaagcacc 5400tgagaacttc cctttgaatg gcttcaataa cagacaagtt gtgctagcgt atgcgaatcc 5460aacgctcagt gccgtttgaa atgatgctca aatgagacag gagtccatct gtataagaag 5520tatggcttaa atggatattt gtcaaattct tacaagatta gtttgtattg atttcaacaa 5580tgctttaacc ttacattgct gctttaaata gttgattaag ctgatcagct tgtaatatgt 5640aatctcttct gggccatcag atccataatg ggtttactag actatataag agaaatagta 5700atattttata aacaattctt gctcagtttt actgtgattt aataacatat gtcattgtgc 5760cctccattgc taagtcaact caactgacga taatactcct tctgaaatag taagaaaaac 5820taatgaagaa cattaattgc tgggtaaaag tgattaattt ctttaaattt gaccagaata 5880atattttgtc agtgaatata ttctcatatc acttgattaa aaacagaaaa ttaccctaac 5940atgaagacca catgtttcct tatcagtctt atcttaattc aagggacaaa aaatctcccc 6000attttagaga tagctagtaa taatcaaccc caaaatgtgg attcggtatg ctccggaact 6060ctccagaaga cagaagacgt ccatctgatg ggattcacac tgagtgggca aaaagttgct 6120gattcccctt tggaggcatc caagcgatgg gctttcagga caggtgtacc tcccaagaat 6180gttgagtaca cagaggggga ggaagccaaa acatgctaca atataagtgt aacggatccc 6240tctggaaaat ccttgctgtt agatcctcct accaacatcc gtgactatcc taaatgcaaa 6300actatccatc atattcaagg tcaaaaccct catgcacagg ggatcgccct tcatttatgg 6360ggagcatttt ttctgtatga tcgcattgcc tccacaacaa tgtaccgagg caaagtcttc 6420actgaaggga acatagcagc tatgattgtc aataagacag tgcacaaaat gattttctcg 6480cggcaaggac aagggtaccg tcatatgaat ctgacttcta ctaataaata ttggacaagt 6540agtaacggaa cgcaaacgaa tgacactgga tgtttcggcg ctcttcaaga atacaattct 6600acaaagaacc aaacatgtgc tccgtccaaa atacctccac cactgcccac agcccgtccg 6660gagatcaaac tcacaagcac cccaactgat gccaccaaac tcaataccac ggacccaagc 6720agtgatgatg aggacctcgc aacatccggc tcagggtccg gagaacgaga accccacaca 6780acttctgatg cggtcaccaa gcaagggctt tcatcaacaa tgccacccac tccctcacca 6840caaccaagca cgccacagca aggaggaaac aacacaaacc attcccaaga tgctgtgact 6900gaactagaca aaaataacac aactgcacaa ccgtccatgc cccctcataa cactaccaca 6960atctctacta acaacacctc caaacacaac ttcagcactc tctctgcacc attacaaaac 7020accaccaatg acaacacaca gagcacaatc actgaaaatg agcaaaccag tgccccctcg 7080ataacaaccc tgcctccaac gggaaatccc accacagcaa agagcaccag cagcaaaaaa 7140ggccccgcca caacggcacc aaacacgaca aatgagcatt tcaccagtcc tccccccacc 7200cccagctcga ctgcacaaca tcttgtatat ttcagaagaa agcgaagtat cctctggagg 7260gaaggcgaca tgttcccttt tctggatggg ttaataaatg ctccaattga ttttgaccca 7320gttccaaata caaaaacaat ctttgatgaa tcctctagtt ctggtgcctc ggctgaggaa 7380gatcaacatg cctcccccaa tattagttta actttatctt attttcctaa tataaatgag 7440aacactgcct actctggaga aaatgagaat gattgtgatg cagagttaag aatttggagc 7500gttcaggagg atgacctggc cgcagggctc agttggatac cgttttttgg ccctggaatt 7560gaaggacttt acactgctgt tttaattaaa aatcaaaaca atttggtctg caggttgagg 7620cgtctagcca atcaaactgc caaatccttg gaactcttat tgagagtcac aactgaggaa 7680agaacattct ccttaatcaa tagacatgct attgactttc tactcacaag atggggagga 7740acatgcaaag tgcttggacc tgattgttgc atcgggatag aagacttgtc caaaaatatt 7800tcagagcaaa ttgaccaaat taaaaaggac gaacaaaaag aggggactgg ttggggtctg 7860ggtggtaaat ggtggacatc cgactggggt gttcttacta acttgggcat tttgctacta 7920ttatccatag ctgtcttgat tgctctatcc tgtatttgtc gtatctttac taaatatatc 7980ggataacgtt aaatgtgtaa tgattaggac tttaggacaa ttgctactga gccctttttc 8040taatctactg aaatcaactt gggagatttt taagaagctg ataacttaat gtgaatcaat 8100agtttatgta ttatcgatta ttatggtttg atattcaatt gttattattg tcaggagtga 8160ccttttctat ttgatgcatt aatgttttaa actacctctt aagcctttga gggcggtccc 8220aatatgtgcg taggggttaa tttaaaggga tttcttattg tacagttttc tgtattactt 8280atttgggctt gaagacatag ttaagatttg ccgaaaatgc tctccagtca attccatccc 8340ctctcagaaa agacgtgctg ttcaaagagt cttaatttat aaccaactat tgcaagaatt 8400aatttacttt ttccgttata cttagttaca ttaatctttt gactgttcag cattattaac 8460gacttgtctt aattcaatcg ttcggatgaa attcataagg aaaaatgagc ctccttcccc 8520ctattctggg ctgagaaaat ttctcttatc cgcctaaaat ctgatctgtt aggtcatggg 8580tccttcataa tctgtttgag catgaatatt gatcaaatga ccaaatgata gtgcatttgt 8640atagactcaa ttatccttta ttaagaaaaa gataaataga acacaaagaa ttgacaaaat 8700tttactttga ttgattttgc aaggagttat aaaaatcttg aaggataaat tgttataaag 8760tagagtcgaa gaacattaaa tgttctttgt tagaattatt catctaagtt gtttttgagt 8820atattcgctt caatacaact cctctttata tttgatttaa gttttaaaat gcaacaacct 8880cgcggaagaa gtcgaacccg caaccaccaa gtcacaccga ctatatatca tgaaactcaa 8940ttgccctcca aacctcatta taccaattat catccacgtg caagatcgat gagctcaacc 9000cgtagtagtg cagaaagtag tcccaccaat catattcccc gtgctcgacc accctcaaca 9060ttcaacttat cgaaaccccc tcctcctcca aaagacatgt gcaggaacat gaaaattgga 9120ttgccgtgcg ctgatcccac ttgtaataga gatcatgacc ttgataatct aacaaatcgt 9180gaacttttgc tattgatggc ccgaaaaatg ctccccaata cagacaagac ctttagaagt 9240ccgcaggact gtggatcacc gtctctttct aaaggtctct caaaagataa acaggagcaa 9300acgaaagatg tgttgacctt ggaaaatcta ggacacattc tgagctatct ccacagatca 9360gaaattggga aattggatga gacatcactt cgtgcagcat taagtctgac gtgtgctgga 9420attcgaaaga cgaatagatc cttgatcaac accatgacag aattacacat gaaccatgaa 9480aatctcccgc aagaccaaaa cggtgttatc aagcagacct atacaggtat tcaccttgac 9540aaaggaggtc aattcgaagc cgccttatgg caaggttggg ataagagatc gatatctcta 9600ttcgtacaag cagctttata tgtaatgaac aatatcccct gtgaatcatc aatcagtgtg 9660caagcctcat acgatcattt tattcttcct caaagtcaag gtaaaggaca gtgattattg 9720ttcgaaagtt gacaatttga tcactttcag ttttcagttt caacccttat cgcgagactt 9780gaatacaatc ctactaactt caataagtga ccccaaattc aagtttgctg aaagctaaga 9840tgacaatgat cactagttca ttgtaaatta ctcgatcaaa atgttcttaa gctatcttaa 9900gcttactgat gcggctctgc ttcacttttc ttttgatttt aaagccatag ctatatctaa 9960gtgtctaatt aacaacttgt acctctaagg aaaaacatga agaacattaa gaaaaaggat 10020gttcttattc tttgactaaa cctgcatatt ctttgttgat accctcgaga gacaactttt 10080gacaccagat cacggatcaa gcacacttca atcaagcacc ctaaattttc aatcatacac 10140ataataacca ttttagtagc gttgcctttc agtacagtct aggtgattgt tgaaagactt 10200ccaagcatgg cagaattatc aacgcgttac aacttgcctg caaatgttac ggaaaatagt 10260ataaatcttg accttaattc cacagcacga tggataaaag aacccagtgt tgggggctgg 10320acagtgaagt ggggaaactt tgttttccat ataccaaata ctggaatgac attgttgcat 10380catttaaagt ctaacttcgt tgttccagag tggcaacaaa caaggaatct attctcccac 10440ctctttaaaa acccaaaatc aacaattata gaaccgtttt tggccctgag gattttgctt 10500ggagttgctt tgaaggatca agaattacag caatcattga ttcctggatt tagatctatt 10560gttcatatgc tatcagaatg gctgctcctg gaggtcacgt cggcaatcca tattagccct 10620aatctgttgg gaatctattt gacttcagac atgtttaaaa ttctgatggc aggtgtgaaa 10680aatttcttca ataagatgtt cactcttcat gttgtaaatg accacggaaa acccagcagt 10740attgaaataa agttaactgg acaacagatc attatcactc gtgttaatat ggggtttcta 10800gtggaagtca ggaggattga tattgaacct tgctgtggtg agacagtcct ctcagaatca 10860gttgtttttg gactagtggc tgaggcagtt ctaagagaac acagtcaaat ggagaagggc 10920caacctctca atctgacaca atacatgaac agcaaaattg ctatataagt ggcttaaatt 10980agcatgggta ttcctagttc gaccacataa taatgttgga ggcacagtac attatagtta 11040attgtcttgt atactaaggg atatacctaa cctgatttat atttactggt ataaaatagt 11100agcatcatct tattgaatag ttatcataca ataggctgtt cctataatct gattgtgaga 11160ttataaactt gtagaattac cgtgggtcac aactgttgca tatcctccaa aatatatctt 11220ttgcaagtga tgtgtgcttg aatacttcga tataatacat actaataacg attgattaag 11280aaaaatcaat gatggatatt aaatgtccat caagcaagtg ttgtagaata ccaggggttt 11340cacaggctgc taaacttact aaattttaca taggattata taattctttt cgatacacgt 11400tatatcttta gcaaagtgag gaaaacagct ttatcatgtt agatgccagt tatccatttt 11460aagtgaatcc tttcctcaac atgcagcatc caactcaata tcctgatgca aggttgtcct 11520ctcctataat cctagaccag tgtgacttat tagccagaag tttagggttg tatagtcatt 11580attcacataa tccgaaattg cgtaattgta ggattccaca tcacatttac cgtttaagga 11640attcgacagc attaaagaca tttcttcaga attgttcgat actcaccgtt ccttttcatt 11700caatttggga tcatatttta acttccattc aatatgatgc aattaatcat gttgatgatt 11760ttaaatacct attgccatct gagctagtca agtatgcgaa ttgggacaac gagttcttaa 11820aagcatatct taataagatc ttaggacttg accatgtttt ttcagcttct gcaaggtcac 11880agtgtgagga tttttctcct aaggaaaatc cttattattg ggggatgtta ttactcgtgc 11940atctatctca acttgccagg aggataaaag gacaaagagg gtcattaaga agtaactgga 12000aatttatagg aacagatttg gagctgtttg gaatagcaga ttttgttatt tttaaggttc 12060cagtaaaaac aataatccga aatgctgtaa gcttacaagc ttcaaaaccg ggattaagaa 12120tatggtaccg tgaccaaaac ttgacccctt atctatgcga cgatgagttt attgtaagcg 12180tcgctagtta tgaatgtttt atcatgatta aagacgtctt cattgagagg tataacacat 12240gggaaatctg tgcccgcgct tggctcgaag acagtgatgg agctgattat cctcctcttg 12300atgtgttagg tgaattatac aaccagggag accaaattat tgccatgtac ctggaggacg 12360gtttcaaatt gattaaacac ttagaaccct tgtgtgtcag ctgtatacaa acacatggca 12420tctttacacc aagaaaatac tggttccaat cacagatgat taagtcatat tatgatgaac 12480tccatgatct caatttgaaa cttcaaattt cagacaataa ggctgagtgt gcccaaaact 12540ttattaaaac tatagttcag gcgaaattga ctcctcaaca atactgtgaa ttattctccc 12600tacaaaagca ttggggtcat cccgttttat acaatgatgt tgcactagat aaggttaaaa 12660aacatgcgca atcgacaaaa atcttaaaac ctaaagtcat gtttgaaact ttttgtgttt 12720tcaaatttat agtagcaaag aatcattatc attctcaagg atcatggtat aaaaccacac 12780atgatttgca tttgactcca tatcttagac aacatattgt gtcaaattca tttccatcac 12840aagccgaaat ttatcagcat ctttgggagt

ggtatttcgt ggagcatgaa cctcttttct 12900caactaaaat aataagtgat ttaagtatct ttataaaaga cagggctacc gctgtgaacc 12960aggagtgttg ggacagtgtc ttcgatagaa gtgtattagg atataaccct cctgttagat 13020ttcaatcaaa gagagtgcca gagcaatttt tgggtcaagc agacttttcc ttgaatcaaa 13080tattagagtt tgctgaaaag ttagagtatt tggctccttc ttataggaat ttttccttct 13140cattaaaaga aaaagagttg aatataggaa gaacttttgg gaagttaccg tatcgtgtca 13200gaaatgtcca aacactcgca gaagccctgc tagcagatgg actggcaaaa gcattcccta 13260gtaacatgat ggttgtcact gagagggaac agaaagaagc attattacat caggcttctt 13320ggcaccacaa ttcagcaagc ataggggaga acgctatagt gaggggtgca agttttgtta 13380ctgatcttga gaaatacaac ctcgccttcc gatatgaatt tacacggcat ttcatagact 13440actgtaatcg atgttatggt gtgaagaatt tatttgattg gatgcatttt ttaataccac 13500tatgttatat gcatgtcagt gacttttata gcccaccaca ttgtgtgaca gaagataatc 13560gaaataaccc acctgattgt gctaatgctt atcattatca cttaggaggt atagagggac 13620ttcagcagaa attgtggaca tgcatatcat gtgcccaaat cacccttgtg gagttaaaaa 13680ctaaattaaa attgaagtcc agcgtcatgg gtgataatca atgtataaca actctaagtc 13740tttttccaat tgatgctccc aacgattatc aagagaacga ggctgaattg aatgcggcac 13800gagttgctgt cgaattagct attactacag gttatagtgg tatattttta aagcctgagg 13860aaacatttgt ccattcaggg ttcatttatt ttggtaaaaa gcaatatctc aacggtgttc 13920aactgccgca atcattgaaa acaatggcaa ggtgtggacc cttatctgac tctattttcg 13980atgatcttca aggttctctg gccagtattg gtacatcctt tgagagagga acaagcgaga 14040cacggcacat ttttccgagc cgttggatag cttcattcca ttcaatgtta gcaataaatt 14100tattaaatca gaatcacctt gggtttcctc tagggttcaa tattgatatt tcttgtttca 14160aaaagcctct taccttctcg gaaaaattaa ttgctctcat aacgccccaa gttttaggag 14220ggttatcatt tttaaatcca gaaaaattgt tctaccggaa cataagtgat cctctcactt 14280caggtctatt tcaactcaag aatgcattgg aatttcttga aaaggaagaa ttattctata 14340tcttgatttc taaaaaacct ggtttagcag atgcctcaga ttttgtcatg aatccattag 14400gcttaaatgt accaggatca aaggaaataa taacgttcct tagacaaaca gttcgcgaaa 14460atatcacgat cacgtcacaa aatagaataa taaattctct tttccacata ggttctgatt 14520tagaggacca aagagtgtgt gagtggcttt tatcatcaaa ccctgtaatg agccgatttg 14580ctgctgacat cttttcaaga acacctagtg gaaaacggct tcaagtctta ggctatctag 14640aaggaacaag aacattacta gcttctcgga caatcagttt aactacagaa ggaacaatgt 14700tgatgaaatt aagagaatta acgagaaacc gatggaaaag ttggttttct tatattgatg 14760cactggacga tgatttatct gagtccttgg aaaagttcac atgtactgtt gatgtggcta 14820atttcttgag ggcatattca tggtctgacg tcttaaaagg gaaaaggcta attggtgcca 14880cactgccatg tttactagag caatttgagg taaagtggat taatttatct gaggatttaa 14940gggaacaatt taatctatct tcagactcaa aatcaactat aaacttgttg ccgtatgact 15000gtaaggaact gcgacttgaa ggaagcaatg acacagagtt aaattatgtc agttgtgctc 15060ttgaccggaa agttgtccag aaacatccct ctgttaatcg tctagcttgg acgataggaa 15120atcgagcacc gtatattggc tcacggacag aagataagat cggttatcct cccttaagag 15180taaattgccc atcagcagca cttaaagaag ctattgagat ggtttctaga ttgttatggg 15240tgactcaagg cactgcagac cgagaaaaat tgcttattcc tcttctcaat tcaagagtaa 15300atctggacta tcagacggtg cttaactttt tacctacaca ctactcaggc aacatagttc 15360atagatataa tgatcaatat ggacaacatt cctttatggc aaacaggatg agtaatacat 15420ctacacgtgc aattatatca actaacacac tgggtaaata tgctggggga ggtcaagctg 15480ctattgatag taatataatc tttcaaaata ctattaattt aggagttgca gttttagata 15540ttgcattgtc tcttgctaaa ttgtcgtcag catcaaatgt cactttccgt ttaatgttaa 15600ataagtgctg cacgcggcat gtaccgtccg aatacctata ttttgataaa cctttagatg 15660tggatttgaa caagtatatg gacaatgagt tggtttatga caatgaccct ctttgcagtg 15720ggattaaagg gagattaggc agagtatctc gatcaacact cacattgagt ttgaatgtca 15780gtgacattgg ttcttatgac tttccaacta ttgctgcatg gacactagga gaaactatag 15840tcggaagcat tttttctgat gaatcttctc aaagtacgga tccaataagc tcaggttgca 15900caaaaacttt cgtcacacat ttccttgtgt atccagttga gagtatattt tatgcattcg 15960gagctaactt aatagttgaa agtttaagtc taagtaggat caaatcaatt aagaacctct 16020cagatttgac attccttata tcatccacaa tcaggaattt atcacataga tcacttcgga 16080ttcttcaatc tactttccga catgaattgg tgctcacccg actagcccac cacataccgt 16140taatttcttt aatgttaggg ggctctgcag gagagaagag ttcatcagat gctgttcggc 16200tatttcttac agcaagttat cagaatttta tcaataattt cagttgtctg atgaagaagg 16260gtcagtcatc gctaccggtt tggctttact ttcctagtga agggcaacaa ttaaagccta 16320tattaaaaat cttacagaga ttatcagact tgttatcacc tgacaaaatt caaaagcgta 16380aaattttggc tgacacctgt tgtccaattg gcagcttttg ggtctatcca agcaagtcca 16440caaggactaa ccattattat gcaagcctta attattggag agacaaagct aataaggtta 16500agaatactcc tttttcacac ttgataaatt gttcatttcc tgaattttct tcacatacca 16560gttcagtctc ctctaatcaa caagtgacca attcgaagta tattgtttat ccagaaaata 16620tcactgaaat aaatgcaaga accagattaa taaattatgg atcaacagct ctacagggga 16680tggacaccaa gatgccactc tcagagcaaa atctagtcga aaattgtcga ccatcagagg 16740gcatcagatt caaggacaat caaaaaataa caaaacatga ccagagatgt gagagggagg 16800aatcttcacc gcaacagatg ttccctgaag ataacatgca gactcctgcg cacatacata 16860gttcctcccc atttcaaatc cttataaaat cactagatgc acatgaggac tttgatgcct 16920cgaagataat cttaaattct gaaataaata atcttaacct tacggagtat actcttaata 16980caaagttatt gacaactcct accaggacag aaattttaga tacaagtccg ttacaatcct 17040ctagatattc atcaacttcc agggaacggt ctctactatc cagagaacaa gcttcatatt 17100tgtacgttga ttgcagtaat attccttcta tctctctaga cccaggtttt cggagtatgt 17160ctgatcagaa tcaagttcaa atgttaatca atacctacaa acgtgattta catgcttgtt 17220ttgatagcaa tcaattctgt cggtttacag gggtagtctc atcaatgcat tacaagcttt 17280atgatctttt gcctccaggt aaattgaaaa aggcaatttg tttggccgaa ggggaaggaa 17340gtggtgctcg gttacttttg aagtggaagg aaacggatta tttattcttc aacactttgg 17400ctacggattc acaacaagaa gccgagattt tgagtggccg ggtaataccg agaatgttgt 17460ataacataga cagattaagt gctttgcttg aatcaaggag actaatattg aacaacctaa 17520ctatccaaat tacagatatt acaaatccat tatggctaga ttctgtaata caatatttac 17580ctgaagatag tgacattctt acaatggacg cagagaccac caaggatgaa acaagggaac 17640agctttataa aactattgtg aatatttgga cacgtacttc tcctaatatc ccaaaaatta 17700gcatcatcaa ggtattttta ttagactatg aagggacttt attcttaatg aagaatgcta 17760ttcagtatta tgggcaagtt caactcaaga aaccatatag ctcaaatgca aaaaactcag 17820aatggtactt gtgttgcggt aaacgaagaa ttcaacggct ccaaattgat ttctcagacc 17880aggtgggaat ttttctgatt tgtaaagcaa tgtcacgcca aagacaagca attccttact 17940ggttaaaaca tatagaaaag aattatcctg cttcattaca cgagtttttc ctaactttgg 18000gtttcccttc tttagagtca tctttctgcc atcgttatac tattccattc agtgaaggaa 18060aggctctttt tcacaaggtc cagtcttatg ttcgtcaagg caaacaacat ttacattctc 18120ttatgttgga ttatgaaaac aattcacctc tactagactt gagaaatcac tttatttgct 18180cattaagggg aaagataact aaatattaca atgatatatt aaagttaaat ctagtcatca 18240aggcagtaga aaaaggtaaa aattggtcac aacttgttga gatccttcct aatatgcatt 18300cagtatgcat agtgcacgtg gatcatgagt gttctggatg cgagaaacgg ttattactta 18360aattggattt tatcagaaat acaaagatcg cagaacaaaa attacttaac agagtaatcg 18420ggtatatcct attctttcca ttcggtctgt ttaaatctgg atcattaagg gcataatttc 18480aacagagaga acttcattta attcacaaaa acaatctatt taagagtgag ggttacattg 18540tctaagatat tgtatgagaa gtaataaaat aaataagaaa acgaaaagac tattagacag 18600cttattttat acaagataat cttatatcgc tttaggcctc acacaagtga gaaaattacg 18660cgcacagatt aactagtgat tagtgtttgg tcacaccaga ggtaactttt taacgttaat 18720tactcagatg ttattgctca taattagcat taatattggc acattgggtg aatccttgag 18780ctttatccct aatatggtgt aagaaattaa ggaaatactg agatacacta gttgaattga 18840attatgacat accatatatc ataaatataa aaaagtgtct gctgtaatct acaagcacct 18900cttttaaata cattaggaaa agaattaagt taccgttgag atcaaaaaaa ccacgtcatg 18960ttttctctga tgacaagtga taaaacttcg tagttaaatt tctagaatgt cgatgtgaat 19020gtaaattaag aaaaaccaat atataaaatt aaaaaattaa aaagctttga tataagtaac 19080acaaaacatt cttcatcttt tttgtgtgtc c 1911126281PRTLake Victoria ebolavirus 26Met Gln Gln Pro Arg Gly Arg Ser Arg Thr Arg Asn His Gln Val Thr1 5 10 15 Pro Thr Ile Tyr His Glu Thr Gln Leu Pro Ser Lys Pro His Tyr Thr 20 25 30 Asn Tyr His Pro Arg Ala Arg Ser Met Ser Ser Thr Arg Ser Ser Ala 35 40 45 Glu Ser Ser Pro Thr Asn His Ile Pro Arg Ala Arg Pro Pro Ser Thr 50 55 60 Phe Asn Leu Ser Lys Pro Pro Pro Pro Pro Lys Asp Met Cys Arg Asn65 70 75 80 Met Lys Ile Gly Leu Pro Cys Ala Asp Pro Thr Cys Asn Arg Asp His 85 90 95 Asp Leu Asp Asn Leu Thr Asn Arg Glu Leu Leu Leu Leu Met Ala Arg 100 105 110 Lys Met Leu Pro Asn Thr Asp Lys Thr Phe Arg Ser Pro Gln Asp Cys 115 120 125 Gly Ser Pro Ser Leu Ser Lys Gly Leu Ser Lys Asp Lys Gln Glu Gln 130 135 140 Thr Lys Asp Val Leu Thr Leu Glu Asn Leu Gly His Ile Leu Ser Tyr145 150 155 160 Leu His Arg Ser Glu Ile Gly Lys Leu Asp Glu Thr Ser Leu Arg Ala 165 170 175 Ala Leu Ser Leu Thr Cys Ala Gly Ile Arg Lys Thr Asn Arg Ser Leu 180 185 190 Ile Asn Thr Met Thr Glu Leu His Met Asn His Glu Asn Leu Pro Gln 195 200 205 Asp Gln Asn Gly Val Ile Lys Gln Thr Tyr Thr Gly Ile His Leu Asp 210 215 220 Lys Gly Gly Gln Phe Glu Ala Ala Leu Trp Gln Gly Trp Asp Lys Arg225 230 235 240 Ser Ile Ser Leu Phe Val Gln Ala Ala Leu Tyr Val Met Asn Asn Ile 245 250 255 Pro Cys Glu Ser Ser Ile Ser Val Gln Ala Ser Tyr Asp His Phe Ile 260 265 270 Leu Pro Gln Ser Gln Gly Lys Gly Gln 275 280 2718959DNAZaire ebolavirus 27cggacacaca aaaagaaaga agaattttta ggatcttttg tgtgcgaata actatgagga 60agattaataa ttttcctctc attgaaattt atatcggaat ttaaattgaa attgttactg 120taatcacacc tggtttgttt cagagccaca tcacaaagat agagaacaac ctaggtctcc 180gaagggagca agggcatcag tgtgctcagt tgaaaatccc ttgtcaacac ctaggtctta 240tcacatcaca agttccacct cagactctgc agggtgatcc aacaacctta atagaaacat 300tattgttaaa ggacagcatt agttcacagt caaacaagca agattgagaa ttaaccttgg 360ttttgaactt gaacacttag gggattgaag attcaacaac cctaaagctt ggggtaaaac 420attggaaata gttaaaagac aaattgctcg gaatcacaaa attccgagta tggattctcg 480tcctcagaaa atctggatgg cgccgagtct cactgaatct gacatggatt accacaagat 540cttgacagca ggtctgtccg ttcaacaggg gattgttcgg caaagagtca tcccagtgta 600tcaagtaaac aatcttgaag aaatttgcca acttatcata caggcctttg aagcaggtgt 660tgattttcaa gagagtgcgg acagtttcct tctcatgctt tgtcttcatc atgcgtacca 720gggagattac aaacttttct tggaaagtgg cgcagtcaag tatttggaag ggcacgggtt 780ccgttttgaa gtcaagaagc gtgatggagt gaagcgcctt gaggaattgc tgccagcagt 840atctagtgga aaaaacatta agagaacact tgctgccatg ccggaagagg agacaactga 900agctaatgcc ggtcagtttc tctcctttgc aagtctattc cttccgaaat tggtagtagg 960agaaaaggct tgccttgaga aggttcaaag gcaaattcaa gtacatgcag agcaaggact 1020gatacaatat ccaacagctt ggcaatcagt aggacacatg atggtgattt tccgtttgat 1080gcgaacaaat tttctgatca aatttctcct aatacaccaa gggatgcaca tggttgccgg 1140gcatgatgcc aacgatgctg tgatttcaaa ttcagtggct caagctcgtt tttcaggctt 1200attgattgtc aaaacagtac ttgatcatat cctacaaaag acagaacgag gagttcgtct 1260ccatcctctt gcaaggaccg ccaaggtaaa aaatgaggtg aactccttta aggctgcact 1320cagctccctg gccaagcatg gagagtatgc tcctttcgcc cgacttttga acctttctgg 1380agtaaataat cttgagcatg gtcttttccc tcaactatcg gcaattgcac tcggagtcgc 1440cacagcacac gggagtaccc tcgcaggagt aaatgttgga gaacagtatc aacaactcag 1500agaggctgcc actgaggctg agaagcaact ccaacaatat gcagagtctc gcgaacttga 1560ccatcttgga cttgatgatc aggaaaagaa aattcttatg aacttccatc agaaaaagaa 1620cgaaatcagc ttccagcaaa caaacgctat ggtaactcta agaaaagagc gcctggccaa 1680gctgacagaa gctatcactg ctgcgtcact gcccaaaaca agtggacatt acgatgatga 1740tgacgacatt ccctttccag gacccatcaa tgatgacgac aatcctggcc atcaagatga 1800tgatccgact gactcacagg atacgaccat tcccgatgtg gtggttgatc ccgatgatgg 1860aagctacggc gaataccaga gttactcgga aaacggcatg aatgcaccag atgacttggt 1920cctattcgat ctagacgagg acgacgagga cactaagcca gtgcctaata gatcgaccaa 1980gggtggacaa cagaagaaca gtcaaaaggg ccagcatata gagggcagac agacacaatc 2040caggccaatt caaaatgtcc caggccctca cagaacaatc caccacgcca gtgcgccact 2100cacggacaat gacagaagaa atgaaccctc cggctcaacc agccctcgca tgctgacacc 2160aattaacgaa gaggcagacc cactggacga tgccgacgac gagacgtcta gccttccgcc 2220cttggagtca gatgatgaag agcaggacag ggacggaact tccaaccgca cacccactgt 2280cgccccaccg gctcccgtat acagagatca ctctgaaaag aaagaactcc cgcaagacga 2340gcaacaagat caggaccaca ctcaagaggc caggaaccag gacagtgaca acacccagtc 2400agaacactct tttgaggaga tgtatcgcca cattctaaga tcacaggggc catttgatgc 2460tgttttgtat tatcatatga tgaaggatga gcctgtagtt ttcagtacca gtgatggcaa 2520agagtacacg tatccagact cccttgaaga ggaatatcca ccatggctca ctgaaaaaga 2580ggctatgaat gaagagaata gatttgttac attggatggt caacaatttt attggccggt 2640gatgaatcac aagaataaat tcatggcaat cctgcaacat catcagtgaa tgagcatgga 2700acaatgggat gattcaaccg acaaatagct aacattaagt agtcaaggaa cgaaaacagg 2760aagaattttt gatgtctaag gtgtgaatta ttatcacaat aaaagtgatt cttatttttg 2820aatttaaagc tagcttatta ttactagccg tttttcaaag ttcaatttga gtcttaatgc 2880aaataggcgt taagccacag ttatagccat aattgtaact caatattcta actagcgatt 2940tatctaaatt aaattacatt atgcttttat aacttaccta ctagcctgcc caacatttac 3000acgatcgttt tataattaag aaaaaactaa tgatgaagat taaaaccttc atcatcctta 3060cgtcaattga attctctagc actcgaagct tattgtcttc aatgtaaaag aaaagctggt 3120ctaacaagat gacaactaga acaaagggca ggggccatac tgcggccacg actcaaaacg 3180acagaatgcc aggccctgag ctttcgggct ggatctctga gcagctaatg accggaagaa 3240ttcctgtaag cgacatcttc tgtgatattg agaacaatcc aggattatgc tacgcatccc 3300aaatgcaaca aacgaagcca aacccgaaga cgcgcaacag tcaaacccaa acggacccaa 3360tttgcaatca tagttttgag gaggtagtac aaacattggc ttcattggct actgttgtgc 3420aacaacaaac catcgcatca gaatcattag aacaacgcat tacgagtctt gagaatggtc 3480taaagccagt ttatgatatg gcaaaaacaa tctcctcatt gaacagggtt tgtgctgaga 3540tggttgcaaa atatgatctt ctggtgatga caaccggtcg ggcaacagca accgctgcgg 3600caactgaggc ttattgggcc gaacatggtc aaccaccacc tggaccatca ctttatgaag 3660aaagtgcgat tcggggtaag attgaatcta gagatgagac cgtccctcaa agtgttaggg 3720aggcattcaa caatctaaac agtaccactt cactaactga ggaaaatttt gggaaacctg 3780acatttcggc aaaggatttg agaaacatta tgtatgatca cttgcctggt tttggaactg 3840ctttccacca attagtacaa gtgatttgta aattgggaaa agatagcaac tcattggaca 3900tcattcatgc tgagttccag gccagcctgg ctgaaggaga ctctcctcaa tgtgccctaa 3960ttcaaattac aaaaagagtt ccaatcttcc aagatgctgc tccacctgtc atccacatcc 4020gctctcgagg tgacattccc cgagcttgcc agaaaagctt gcgtccagtc ccaccatcgc 4080ccaagattga tcgaggttgg gtatgtgttt ttcagcttca agatggtaaa acacttggac 4140tcaaaatttg agccaatctc ccttccctcc gaaagaggcg aataatagca gaggcttcaa 4200ctgctgaact atagggtacg ttacattaat gatacacttg tgagtatcag ccctggataa 4260tataagtcaa ttaaacgacc aagataaaat tgttcatatc tcgctagcag cttaaaatat 4320aaatgtaata ggagctatat ctctgacagt attataatca attgttatta agtaacccaa 4380accaaaagtg atgaagatta agaaaaacct acctcggctg agagagtgtt ttttcattaa 4440ccttcatctt gtaaacgttg agcaaaattg ttaaaaatat gaggcgggtt atattgccta 4500ctgctcctcc tgaatatatg gaggccatat accctgtcag gtcaaattca acaattgcta 4560gaggtggcaa cagcaataca ggcttcctga caccggagtc agtcaatggg gacactccat 4620cgaatccact caggccaatt gccgatgaca ccatcgacca tgccagccac acaccaggca 4680gtgtgtcatc agcattcatc cttgaagcta tggtgaatgt catatcgggc cccaaagtgc 4740taatgaagca aattccaatt tggcttcctc taggtgtcgc tgatcaaaag acctacagct 4800ttgactcaac tacggccgcc atcatgcttg cttcatacac tatcacccat ttcggcaagg 4860caaccaatcc acttgtcaga gtcaatcggc tgggtcctgg aatcccggat catcccctca 4920ggctcctgcg aattggaaac caggctttcc tccaggagtt cgttcttccg ccagtccaac 4980taccccagta tttcaccttt gatttgacag cactcaaact gatcacccaa ccactgcctg 5040ctgcaacatg gaccgatgac actccaacag gatcaaatgg agcgttgcgt ccaggaattt 5100catttcatcc aaaacttcgc cccattcttt tacccaacaa aagtgggaag aaggggaaca 5160gtgccgatct aacatctccg gagaaaatcc aagcaataat gacttcactc caggacttta 5220agatcgttcc aattgatcca accaaaaata tcatgggaat cgaagtgcca gaaactctgg 5280tccacaagct gaccggtaag aaggtgactt ctaaaaatgg acaaccaatc atccctgttc 5340ttttgccaaa gtacattggg ttggacccgg tggctccagg agacctcacc atggtaatca 5400cacaggattg tgacacgtgt cattctcctg caagtcttcc agctgtgatt gagaagtaat 5460tgcaataatt gactcagatc cagttttata gaatcttctc agggatagtg ataacatcta 5520tttagtaatc cgtccattag aggagacact tttaattgat caatatacta aaggtgcttt 5580acaccattgt cttttttctc tcctaaatgt agaacttaac aaaagactca taatatactt 5640gtttttaaag gattgattga tgaaagatca taactaataa cattacaaat aatcctacta 5700taatcaatac ggtgattcaa atgttaatct ttctcattgc acatactttt tgcccttatc 5760ctcaaattgc ctgcatgctt acatctgagg atagccagtg tgacttggat tggaaatgtg 5820gagaaaaaat cgggacccat ttctaggttg ttcacaatcc aagtacagac attgcccttc 5880taattaagaa aaaatcggcg atgaagatta agccgacagt gagcgtaatc ttcatctctc 5940ttagattatt tgttttccag agtaggggtc gtcaggtcct tttcaatcgt gtaaccaaaa 6000taaactccac tagaaggata ttgtggggca acaacacaat gggcgttaca ggaatattgc 6060agttacctcg tgatcgattc aagaggacat cattctttct ttgggtaatt atccttttcc 6120aaagaacatt ttccatccca cttggagtca tccacaatag cacattacag gttagtgatg 6180tcgacaaact agtttgtcgt gacaaactgt catccacaaa tcaattgaga tcagttggac 6240tgaatctcga agggaatgga gtggcaactg acgtgccatc tgcaactaaa agatggggct 6300tcaggtccgg tgtcccacca aaggtggtca attatgaagc tggtgaatgg gctgaaaact 6360gctacaatct tgaaatcaaa aaacctgacg ggagtgagtg tctaccagca gcgccagacg 6420ggattcgggg cttcccccgg tgccggtatg tgcacaaagt atcaggaacg ggaccgtgtg 6480ccggagactt tgccttccat aaagagggtg ctttcttcct gtatgatcga cttgcttcca 6540cagttatcta ccgaggaacg actttcgctg aaggtgtcgt tgcatttctg atactgcccc 6600aagctaagaa ggacttcttc agctcacacc ccttgagaga gccggtcaat gcaacggagg 6660acccgtctag tggctactat tctaccacaa ttagatatca ggctaccggt tttggaacca 6720atgagacaga gtacttgttc gaggttgaca atttgaccta cgtccaactt gaatcaagat 6780tcacaccaca gtttctgctc cagctgaatg agacaatata tacaagtggg aaaaggagca 6840ataccacggg aaaactaatt tggaaggtca accccgaaat tgatacaaca atcggggagt 6900gggccttctg

ggaaactaaa aaaacctcac tagaaaaatt cgcagtgaag agttgtcttt 6960cacagttgta tcaaacggag ccaaaaacat cagtggtcag agtccggcgc gaacttcttc 7020cgacccaggg accaacacaa caactgaaga ccacaaaatc atggcttcag aaaattcctc 7080tgcaatggtt caagtgcaca gtcaaggaag ggaagctgca gtgtcgcatc taacaaccct 7140tgccacaatc tccacgagtc cccaatccct cacaaccaaa ccaggtccgg acaacagcac 7200ccataataca cccgtgtata aacttgacat ctctgaggca actcaagttg aacaacatca 7260ccgcagaaca gacaacgaca gcacagcctc cgacactccc tctgccacga ccgcagccgg 7320acccccaaaa gcagagaaca ccaacacgag caagagcact gacttcctgg accccgccac 7380cacaacaagt ccccaaaacc acagcgagac cgctggcaac aacaacactc atcaccaaga 7440taccggagaa gagagtgcca gcagcgggaa gctaggctta attaccaata ctattgctgg 7500agtcgcagga ctgatcacag gcgggagaag aactcgaaga gaagcaattg tcaatgctca 7560acccaaatgc aaccctaatt tacattactg gactactcag gatgaaggtg ctgcaatcgg 7620actggcctgg ataccatatt tcgggccagc agccgaggga atttacatag aggggctaat 7680gcacaatcaa gatggtttaa tctgtgggtt gagacagctg gccaacgaga cgactcaagc 7740tcttcaactg ttcctgagag ccacaactga gctacgcacc ttttcaatcc tcaaccgtaa 7800ggcaattgat ttcttgctgc agcgatgggg cggcacatgc cacattctgg gaccggactg 7860ctgtatcgaa ccacatgatt ggaccaagaa cataacagac aaaattgatc agattattca 7920tgattttgtt gataaaaccc ttccggacca gggggacaat gacaattggt ggacaggatg 7980gagacaatgg ataccggcag gtattggagt tacaggcgtt ataattgcag ttatcgcttt 8040attctgtata tgcaaatttg tcttttagtt tttcttcaga ttgcttcatg gaaaagctca 8100gcctcaaatc aatgaaacca ggatttaatt atatggatta cttgaatcta agattacttg 8160acaaatgata atataataca ctggagcttt aaacatagcc aatgtgattc taactccttt 8220aaactcacag ttaatcataa acaaggtttg acatcaatct agttatctct ttgagaatga 8280taaacttgat gaagattaag aaaaaggtaa tctttcgatt atctttaatc ttcatccttg 8340attctacaat catgacagtt gtctttagtg acaagggaaa gaagcctttt tattaagttg 8400taataatcag atctgcgaac cggtagagtt tagttgcaac ctaacacaca taaagcattg 8460gtcaaaaagt caatagaaat ttaaacagtg agtggagaca acttttaaat ggaagcttca 8520tatgagagag gacgcccacg agctgccaga cagcattcaa gggatggaca cgaccaccat 8580gttcgagcac gatcatcatc cagagagaat tatcgaggtg agtaccgtca atcaaggagc 8640gcctcacaag tgcgcgttcc tactgtattt cataagaaga gagttgaacc attaacagtt 8700cctccagcac ctaaagacat atgtccgacc ttgaaaaaag gatttttgtg tgacagtagt 8760ttttgcaaaa aagatcacca gttggagagt ttaactgata gggaattact cctactaatc 8820gcccgtaaga cttgtggatc agtagaacaa caattaaata taactgcacc caaggactcg 8880cgcttagcaa atccaacggc tgatgatttc cagcaagagg aaggtccaaa aattaccttg 8940ttgacactga tcaagacggc agaacactgg gcgagacaag acatcagaac catagaggat 9000tcaaaattaa gagcattgtt gactctatgt gctgtgatga cgaggaaatt ctcaaaatcc 9060cagctgagtc ttttatgtga gacacaccta aggcgcgagg ggcttgggca agatcaggca 9120gaacccgttc tcgaagtata tcaacgatta cacagtgata aaggaggcag ttttgaagct 9180gcactatggc aacaatggga ccgacaatcc ctaattatgt ttatcactgc attcttgaat 9240attgctctcc agttaccgtg tgaaagttct gctgtcgttg tttcagggtt aagaacattg 9300gttcctcaat cagataatga ggaagcttca accaacccgg ggacatgctc atggtctgat 9360gagggtaccc cttaataagg ctgactaaaa cactatataa ccttctactt gatcacaata 9420ctccgtatac ctatcatcat atatttaatc aagacgatat cctttaaaac ttattcagta 9480ctataatcac tctcgtttca aattaataag atgtgcatga ttgccctaat atatgaagag 9540gtatgataca accctaacag tgatcaaaga aaatcataat ctcgtatcgc tcgtaatata 9600acctgccaag catacctctt gcacaaagtg attcttgtac acaaataatg ttttactcta 9660caggaggtag caacgatcca tcccatcaaa aaataagtat ttcatgactt actaatgatc 9720tcttaaaata ttaagaaaaa ctgacggaac ataaattctt tatgcttcaa gctgtggagg 9780aggtgtttgg tattggctat tgttatatta caatcaataa caagcttgta aaaatattgt 9840tcttgtttca agaggtagat tgtgaccgga aatgctaaac taatgatgaa gattaatgcg 9900gaggtctgat aagaataaac cttattattc agattaggcc ccaagaggca ttcttcatct 9960ccttttagca aagtactatt tcagggtagt ccaattagtg gcacgtcttt tagctgtata 10020tcagtcgccc ctgagatacg ccacaaaagt gtctctaagc taaattggtc tgtacacatc 10080ccatacattg tattaggggc aataatatct aattgaactt agccgtttaa aatttagtgc 10140ataaatctgg gctaacacca ccaggtcaac tccattggct gaaaagaagc ttacctacaa 10200cgaacatcac tttgagcgcc ctcacaatta aaaaatagga acgtcgttcc aacaatcgag 10260cgcaaggttt caaggttgaa ctgagagtgt ctagacaaca aaatattgat actccagaca 10320ccaagcaaga cctgagaaaa aaccatggct aaagctacgg gacgatacaa tctaatatcg 10380cccaaaaagg acctggagaa aggggttgtc ttaagcgacc tctgtaactt cttagttagc 10440caaactattc aggggtggaa ggtttattgg gctggtattg agtttgatgt gactcacaaa 10500ggaatggccc tattgcatag actgaaaact aatgactttg cccctgcatg gtcaatgaca 10560aggaatctct ttcctcattt atttcaaaat ccgaattcca caattgaatc accgctgtgg 10620gcattgagag tcatccttgc agcagggata caggaccagc tgattgacca gtctttgatt 10680gaacccttag caggagccct tggtctgatc tctgattggc tgctaacaac caacactaac 10740catttcaaca tgcgaacaca acgtgtcaag gaacaattga gcctaaaaat gctgtcgttg 10800attcgatcca atattctcaa gtttattaac aaattggatg ctctacatgt cgtgaactac 10860aacggattgt tgagcagtat tgaaattgga actcaaaatc atacaatcat cataactcga 10920actaacatgg gttttctggt ggagctccaa gaacccgaca aatcggcaat gaaccgcatg 10980aagcctgggc cggcgaaatt ttccctcctt catgagtcca cactgaaagc atttacacaa 11040ggatcctcga cacgaatgca aagtttgatt cttgaattta atagctctct tgctatctaa 11100ctaaggtaga atacttcata ttgagctaac tcatatatgc tgactcaata gttatcttga 11160catctctgct ttcataatca gatatataag cataataaat aaatactcat atttcttgat 11220aatttgttta accacagata aatcctcact gtaagccagc ttccaagttg acacccttac 11280aaaaaccagg actcagaatc cctcaaacaa gagattccaa gacaacatca tagaattgct 11340ttattatatg aataagcatt ttatcaccag aaatcctata tactaaatgg ttaattgtaa 11400ctgaacccgc aggtcacatg tgttaggttt cacagattct atatattact aactctatac 11460tcgtaattaa cattagataa gtagattaag aaaaaagcct gaggaagatt aagaaaaact 11520gcttattggg tctttccgtg ttttagatga agcagttgaa attcttcctc ttgatattaa 11580atggctacac aacataccca atacccagac gctaggttat catcaccaat tgtattggac 11640caatgtgacc tagtcactag agcttgcggg ttatattcat catactccct taatccgcaa 11700ctacgcaact gtaaactccc gaaacatatc taccgtttga aatacgatgt aactgttacc 11760aagttcttga gtgatgtacc agtggcgaca ttgcccatag atttcatagt cccagttctt 11820ctcaaggcac tgtcaggcaa tggattctgt cctgttgagc cgcggtgcca acagttctta 11880gatgaaatca ttaagtacac aatgcaagat gctctcttct tgaaatatta tctcaaaaat 11940gtgggtgctc aagaagactg tgttgatgaa cactttcaag agaaaatctt atcttcaatt 12000cagggcaatg aatttttaca tcaaatgttt ttctggtatg atctggctat tttaactcga 12060aggggtagat taaatcgagg aaactctaga tcaacatggt ttgttcatga tgatttaata 12120gacatcttag gctatgggga ctatgttttt tggaagatcc caatttcaat gttaccactg 12180aacacacaag gaatccccca tgctgctatg gactggtatc aggcatcagt attcaaagaa 12240gcggttcaag ggcatacaca cattgtttct gtttctactg ccgacgtctt gataatgtgc 12300aaagatttaa ttacatgtcg attcaacaca actctaatct caaaaatagc agagattgag 12360gatccagttt gttctgatta tcccaatttt aagattgtgt ctatgcttta ccagagcgga 12420gattacttac tctccatatt agggtctgat gggtataaaa ttattaagtt cctcgaacca 12480ttgtgcttgg ccaaaattca attatgctca aagtacactg agaggaaggg ccgattctta 12540acacaaatgc atttagctgt aaatcacacc ctagaagaaa ttacagaaat gcgtgcacta 12600aagccttcac aggctcaaaa gatccgtgaa ttccatagaa cattgataag gctggagatg 12660acgccacaac aactttgtga gctattttcc attcaaaaac actgggggca tcctgtgcta 12720catagtgaaa cagcaatcca aaaagttaaa aaacatgcta cggtgctaaa agcattacgc 12780cctatagtga ttttcgagac atactgtgtt tttaaatata gtattgccaa acattatttt 12840gatagtcaag gatcttggta cagtgttact tcagatagga atctaacacc gggtcttaat 12900tcttatatca aaagaaatca attccctccg ttgccaatga ttaaagaact actatgggaa 12960ttttaccacc ttgaccaccc tccacttttc tcaaccaaaa ttattagtga cttaagtatt 13020tttataaaag acagagctac cgcagtagaa aggacatgct gggatgcagt attcgagcct 13080aatgttctag gatataatcc acctcacaaa tttagtacta aacgtgtacc ggaacaattt 13140ttagagcaag aaaacttttc tattgagaat gttctttcct acgcacaaaa actcgagtat 13200ctactaccac aatatcggaa cttttctttc tcattgaaag agaaagagtt gaatgtaggt 13260agaaccttcg gaaaattgcc ttatccgact cgcaatgttc aaacactttg tgaagctctg 13320ttagctgatg gtcttgctaa agcatttcct agcaatatga tggtagttac ggaacgtgag 13380caaaaagaaa gcttattgca tcaagcatca tggcaccaca caagtgatga ttttggtgaa 13440catgccacag ttagagggag tagctttgta actgatttag agaaatacaa tcttgcattt 13500agatatgagt ttacagcacc ttttatagaa tattgcaacc gttgctatgg tgttaagaat 13560gtttttaatt ggatgcatta tacaatccca cagtgttata tgcatgtcag tgattattat 13620aatccaccac ataacctcac actggagaat cgagacaacc cccccgaagg gcctagttca 13680tacaggggtc atatgggagg gattgaagga ctgcaacaaa aactctggac aagtatttca 13740tgtgctcaaa tttctttagt tgaaattaag actggtttta agttacgctc agctgtgatg 13800ggtgacaatc agtgcattac tgttttatca gtcttcccct tagagactga cgcagacgag 13860caggaacaga gcgccgaaga caatgcagcg agggtggccg ccagcctagc aaaagttaca 13920agtgcctgtg gaatcttttt aaaacctgat gaaacatttg tacattcagg ttttatctat 13980tttggaaaaa aacaatattt gaatggggtc caattgcctc agtcccttaa aacggctaca 14040agaatggcac cattgtctga tgcaattttt gatgatcttc aagggaccct ggctagtata 14100ggcactgctt ttgagcgatc catctctgag acacgacata tctttccttg caggataacc 14160gcagctttcc atacgttttt ttcggtgaga atcttgcaat atcatcatct cgggttcaat 14220aaaggttttg accttggaca gttaacactc ggcaaacctc tggatttcgg aacaatatca 14280ttggcactag cggtaccgca ggtgcttgga gggttatcct tcttgaatcc tgagaaatgt 14340ttctaccgga atctaggaga tccagttacc tcaggcttat tccagttaaa aacttatctc 14400cgaatgattg agatggatga tttattctta cctttaattg cgaagaaccc tgggaactgc 14460actgccattg actttgtgct aaatcctagc ggattaaatg tccctgggtc gcaagactta 14520acttcatttc tgcgccagat tgtacgcagg accatcaccc taagtgcgaa aaacaaactt 14580attaatacct tatttcatgc gtcagctgac ttcgaagacg aaatggtttg taaatggcta 14640ttatcatcaa ctcctgttat gagtcgtttt gcggccgata tcttttcacg cacgccgagc 14700gggaagcgat tgcaaattct aggatacctg gaaggaacac gcacattatt agcctctaag 14760atcatcaaca ataatacaga gacaccggtt ttggacagac tgaggaaaat aacattgcaa 14820aggtggagcc tatggtttag ttatcttgat cattgtgata atatcctggc ggaggcttta 14880acccaaataa cttgcacagt tgatttagca cagattctga gggaatattc atgggctcat 14940attttagagg gaagacctct tattggagcc acactcccat gtatgattga gcaattcaaa 15000gtgttttggc tgaaacccta cgaacaatgt ccgcagtgtt caaatgcaaa gcaaccaggt 15060gggaaaccat tcgtgtcagt ggcagtcaag aaacatattg ttagtgcatg gccgaacgca 15120tcccgaataa gctggactat cggggatgga atcccataca ttggatcaag gacagaagat 15180aagataggac aacctgctat taaaccaaaa tgtccttccg cagccttaag agaggccatt 15240gaattggcgt cccgtttaac atgggtaact caaggcagtt cgaacagtga cttgctaata 15300aaaccatttt tggaagcacg agtaaattta agtgttcaag aaatacttca aatgacccct 15360tcacattact caggaaatat tgttcacagg tacaacgatc aatacagtcc tcattctttc 15420atggccaatc gtatgagtaa ttcagcaacg cgattgattg tttctacaaa cactttaggt 15480gagttttcag gaggtggcca gtctgcacgc gacagcaata ttattttcca gaatgttata 15540aattatgcag ttgcactgtt cgatattaaa tttagaaaca ctgaggctac agatatccaa 15600tataatcgtg ctcaccttca tctaactaag tgttgcaccc gggaagtacc agctcagtat 15660ttaacataca catctacatt ggatttagat ttaacaagat accgagaaaa cgaattgatt 15720tatgacagta atcctctaaa aggaggactc aattgcaata tctcattcga taatccattt 15780ttccaaggta aacggctgaa cattatagaa gatgatctta ttcgactgcc tcacttatct 15840ggatgggagc tagccaagac catcatgcaa tcaattattt cagatagcaa caattcatct 15900acagacccaa ttagcagtgg agaaacaaga tcattcacta cccatttctt aacttatccc 15960aagataggac ttctgtacag ttttggggcc tttgtaagtt attatcttgg caatacaatt 16020cttcggacta agaaattaac acttgacaat tttttatatt acttaactac tcaaattcat 16080aatctaccac atcgctcatt gcgaatactt aagccaacat tcaaacatgc aagcgttatg 16140tcacggttaa tgagtattga tcctcatttt tctatttaca taggcggtgc tgcaggtgac 16200agaggactct cagatgcggc caggttattt ttgagaacgt ccatttcatc ttttcttaca 16260tttgtaaaag aatggataat taatcgcgga acaattgtcc ctttatggat agtatatccg 16320ctagagggtc aaaacccaac acctgtgaat aattttctct atcagatcgt agaactgctg 16380gtgcatgatt catcaagaca acaggctttt aaaactacca taagtgatca tgtacatcct 16440cacgacaatc ttgtttacac atgtaagagt acagccagca atttcttcca tgcatcattg 16500gcgtactgga ggagcagaca cagaaacagc aaccgaaaat acttggcaag agactcttca 16560actggatcaa gcacaaacaa cagtgatggt catattgaga gaagtcaaga acaaaccacc 16620agagatccac atgatggcac tgaacggaat ctagtcctac aaatgagcca tgaaataaaa 16680agaacgacaa ttccacaaga aaacacgcac cagggtccgt cgttccagtc ctttctaagt 16740gactctgctt gtggtacagc aaatccaaaa ctaaatttcg atcgatcgag acacaatgtg 16800aaatttcagg atcataactc ggcatccaag agggaaggtc atcaaataat ctcacaccgt 16860ctagtcctac ctttctttac attatctcaa gggacacgcc aattaacgtc atccaatgag 16920tcacaaaccc aagacgagat atcaaagtac ttacggcaat tgagatccgt cattgatacc 16980acagtttatt gtagatttac cggtatagtc tcgtccatgc attacaaact tgatgaggtc 17040ctttgggaaa tagagagttt caagtcggct gtgacgctag cagagggaga aggtgctggt 17100gccttactat tgattcagaa ataccaagtt aagaccttat ttttcaacac gctagctact 17160gagtccagta tagagtcaga aatagtatca ggaatgacta ctcctaggat gcttctacct 17220gttatgtcaa aattccataa tgaccaaatt gagattattc ttaacaactc agcaagccaa 17280ataacagaca taacaaatcc tacttggttt aaagaccaaa gagcaaggct acctaagcaa 17340gtcgaggtta taaccatgga tgcagagaca acagagaata taaacagatc gaaattgtac 17400gaagctgtat ataaattgat cttacaccat attgatccta gcgtattgaa agcagtggtc 17460cttaaagtct ttctaagtga tactgagggt atgttatggc taaatgataa tttagccccg 17520ttttttgcca ctggttattt aattaagcca ataacgtcaa gtgctagatc tagtgagtgg 17580tatctttgtc tgacgaactt cttatcaact acacgtaaga tgccacacca aaaccatctc 17640agttgtaaac aggtaatact tacggcattg caactgcaaa ttcaacgaag cccatactgg 17700ctaagtcatt taactcagta tgctgactgt gagttacatt taagttatat ccgccttggt 17760tttccatcat tagagaaagt actataccac aggtataacc tcgtcgattc aaaaagaggt 17820ccactagtct ctatcactca gcacttagca catcttagag cagagattcg agaattaact 17880aatgattata atcaacagcg acaaagtcgg actcaaacat atcactttat tcgtactgca 17940aaaggacgaa tcacaaaact agtcaatgat tatttaaaat tctttcttat tgtgcaagca 18000ttaaaacata atgggacatg gcaagctgag tttaagaaat taccagagtt gattagtgtg 18060tgcaataggt tctaccatat tagagattgc aattgtgaag aacgtttctt agttcaaacc 18120ttatatttac atagaatgca ggattctgaa gttaagctta tcgaaaggct gacagggctt 18180ctgagtttat ttccggatgg tctctacagg tttgattgaa ttaccgtgca tagtatcctg 18240atacttgcaa aggttggtta ttaacataca gattataaaa aactcataaa ttgctctcat 18300acatcatatt gatctaatct caataaacaa ctatttaaat aacgaaagga gtccctatat 18360tatatactat atttagcctc tctccctgcg tgataatcaa aaaattcaca atgcagcatg 18420tgtgacatat tactgccgca atgaatttaa cgcaacataa taaactctgc actctttata 18480attaagcttt aacgaaaggt ctgggctcat attgttattg atataataat gttgtatcaa 18540tatcctgtca gatggaatag tgttttggtt gataacacaa cttcttaaaa caaaattgat 18600ctttaagatt aagtttttta taattatcat tactttaatt tgtcgtttta aaaacggtga 18660tagccttaat ctttgtgtaa aataagagat taggtgtaat aaccttaaca tttttgtcta 18720gtaagctact atttcataca gaatgataaa attaaaagaa aaggcaggac tgtaaaatca 18780gaaatacctt ctttacaata tagcagacta gataataatc ttcgtgttaa tgataattaa 18840gacattgacc acgctcatca gaaggctcgc cagaataaac gttgcaaaaa ggattcctgg 18900aaaaatggtc gcacacaaaa atttaaaaat aaatctattt cttctttttt gtgtgtcca 1895928288PRTZaire ebolavirus 28Met Glu Ala Ser Tyr Glu Arg Gly Arg Pro Arg Ala Ala Arg Gln His1 5 10 15 Ser Arg Asp Gly His Asp His His Val Arg Ala Arg Ser Ser Ser Arg 20 25 30 Glu Asn Tyr Arg Gly Glu Tyr Arg Gln Ser Arg Ser Ala Ser Gln Val 35 40 45 Arg Val Pro Thr Val Phe His Lys Lys Arg Val Glu Pro Leu Thr Val 50 55 60 Pro Pro Ala Pro Lys Asp Ile Cys Pro Thr Leu Lys Lys Gly Phe Leu65 70 75 80 Cys Asp Ser Ser Phe Cys Lys Lys Asp His Gln Leu Glu Ser Leu Thr 85 90 95 Asp Arg Glu Leu Leu Leu Leu Ile Ala Arg Lys Thr Cys Gly Ser Val 100 105 110 Glu Gln Gln Leu Asn Ile Thr Ala Pro Lys Asp Ser Arg Leu Ala Asn 115 120 125 Pro Thr Ala Asp Asp Phe Gln Gln Glu Glu Gly Pro Lys Ile Thr Leu 130 135 140 Leu Thr Leu Ile Lys Thr Ala Glu His Trp Ala Arg Gln Asp Ile Arg145 150 155 160 Thr Ile Glu Asp Ser Lys Leu Arg Ala Leu Leu Thr Leu Cys Ala Val 165 170 175 Met Thr Arg Lys Phe Ser Lys Ser Gln Leu Ser Leu Leu Cys Glu Thr 180 185 190 His Leu Arg Arg Glu Gly Leu Gly Gln Asp Gln Ala Glu Pro Val Leu 195 200 205 Glu Val Tyr Gln Arg Leu His Ser Asp Lys Gly Gly Ser Phe Glu Ala 210 215 220 Ala Leu Trp Gln Gln Trp Asp Arg Gln Ser Leu Ile Met Phe Ile Thr225 230 235 240 Ala Phe Leu Asn Ile Ala Leu Gln Leu Pro Cys Glu Ser Ser Ala Val 245 250 255 Val Val Ser Gly Leu Arg Thr Leu Val Pro Gln Ser Asp Asn Glu Glu 260 265 270 Ala Ser Thr Asn Pro Gly Thr Cys Ser Trp Ser Asp Glu Gly Thr Pro 275 280 285 2918959DNAZaire ebolavirus 29cggacacaca aaaagaaaga agaattttta ggatcttttg tgtgcgaata actatgagga 60agattaataa ttttcctctc attgaaattt atatcggaat ttaaattgaa attgttactg 120taatcacacc tggtttgttt cagagccaca tcacaaagat agagaacaac ctaggtctcc 180gaagggagca agggcatcag tgtgctcagt tgaaaatccc ttgtcaacac ctaggtctta 240tcacatcaca agttccacct cagactctgc agggtgatcc aacaacctta atagaaacat 300tattgttaaa ggacagcatt agttcacagt caaacaagca agattgagaa ttaaccttgg 360ttttgaactt gaacacttag gggattgaag attcaacaac cctaaagctt ggggtaaaac 420attggaaata gttaaaagac aaattgctcg gaatcacaaa attccgagta tggattctcg 480tcctcagaaa atctggatgg cgccgagtct cactgaatct gacatggatt accacaagat 540cttgacagca ggtctgtccg ttcaacaggg gattgttcgg caaagagtca tcccagtgta 600tcaagtaaac aatcttgaag aaatttgcca acttatcata caggcctttg aagcaggtgt 660tgattttcaa gagagtgcgg acagtttcct tctcatgctt tgtcttcatc atgcgtacca 720gggagattac aaacttttct tggaaagtgg cgcagtcaag tatttggaag ggcacgggtt 780ccgttttgaa gtcaagaagc gtgatggagt gaagcgcctt gaggaattgc tgccagcagt 840atctagtgga aaaaacatta agagaacact tgctgccatg ccggaagagg agacaactga 900agctaatgcc ggtcagtttc tctcctttgc aagtctattc cttccgaaat tggtagtagg 960agaaaaggct tgccttgaga aggttcaaag gcaaattcaa gtacatgcag agcaaggact 1020gatacaatat ccaacagctt ggcaatcagt aggacacatg atggtgattt tccgtttgat

1080gcgaacaaat tttctgatca aatttctcct aatacaccaa gggatgcaca tggttgccgg 1140gcatgatgcc aacgatgctg tgatttcaaa ttcagtggct caagctcgtt tttcaggctt 1200attgattgtc aaaacagtac ttgatcatat cctacaaaag acagaacgag gagttcgtct 1260ccatcctctt gcaaggaccg ccaaggtaaa aaatgaggtg aactccttta aggctgcact 1320cagctccctg gccaagcatg gagagtatgc tcctttcgcc cgacttttga acctttctgg 1380agtaaataat cttgagcatg gtcttttccc tcaactatcg gcaattgcac tcggagtcgc 1440cacagcacac gggagtaccc tcgcaggagt aaatgttgga gaacagtatc aacaactcag 1500agaggctgcc actgaggctg agaagcaact ccaacaatat gcagagtctc gcgaacttga 1560ccatcttgga cttgatgatc aggaaaagaa aattcttatg aacttccatc agaaaaagaa 1620cgaaatcagc ttccagcaaa caaacgctat ggtaactcta agaaaagagc gcctggccaa 1680gctgacagaa gctatcactg ctgcgtcact gcccaaaaca agtggacatt acgatgatga 1740tgacgacatt ccctttccag gacccatcaa tgatgacgac aatcctggcc atcaagatga 1800tgatccgact gactcacagg atacgaccat tcccgatgtg gtggttgatc ctgatgatgg 1860aagctacggc gaataccaga gttactcgga aaacggcatg aatgcaccag atgacttggt 1920cctattcgat ctagacgagg acgacgagga cactaagcca gtgcctaata gatcgaccaa 1980gggtggacaa cagaagaaca gtcaaaaggg ccagcatata gagggcagac agacacaatt 2040caggccaatt caaaatgtcc caggccctca cagaacaatc caccacgcca gtgcgccact 2100cacggacaat gacagaagaa atgaaccctc cggctcaacc agccctcgca tgctgacacc 2160aattaacgaa gaggcagacc cactggacga tgccgacgac gagacgtcta gccttccgcc 2220cttggagtca gatgatgaag agcaggacag ggacggaact tccaaccgca cacccactgt 2280cgccccaccg gctcccgtat acagagatca ctctgaaaag aaagaactcc cgcaagacga 2340gcaacaagat caggaccaca ctcaagaggc caggaaccag gacagtgaca acacccagtc 2400agaacactcc cttgaggaga tgtatcgcca cattctaaga tcacaggggc catttgatgc 2460tgttttgtat tatcatatga tgaaggatga gcctgtagtt ttcagtacca gtgatggcaa 2520agagtacacg tatccagact cccttgaaga ggaatatcca ccatggctca ctgaaaaaga 2580ggctatgaat gaagagaata gatttgttac attggatggt caacaatttt attggccggt 2640gatgaatcac aagaataaat tcatggcaat cctgcaacat catcagtgaa tgagcatgga 2700acaatgggat gattcaaccg acaaatagct aacattaagt agtcaaggaa cgaaaacagg 2760aagaattttt gatgtctaag gtgtgaatta ttatcacaat aaaagtgatt cttatttttg 2820aatttaaagc tagcttatta ttactagccg tttttcaaag ttcaatttga gtcttaatgc 2880aaataggcgt taagccacag ttatagccat aattgtaact caatattcta actagcgatt 2940tatctaaatt aaattacatt atgcttttat aacttaccta ctagcctgcc caacatttac 3000acgatcgttt tataattaag aaaaaactaa tgatgaagat taaaaccttc atcatcctta 3060cgtcaattga attctctagc actcgaagct tattgtcttc aatgtaaaag aaaagctggt 3120ctaacaagat gacaactaga acaaagggca ggggccatac tgcggccacg actcaaaacg 3180acagaatgcc aggccctgag ctttcgggct ggatctctga gcagctaatg accggaagaa 3240ttcctgtaag cgacatcttc tgtgatattg agaacaatcc aggattatgc tacgcatccc 3300aaatgcaaca aacgaagcca aacccgaaga cgcgcaacag tcaaacccaa acggacccaa 3360tttgcaatca tagttttgag gaggtagtac aaacattggc ttcattggct actgttgtgc 3420aacaacaaac catcgcatca gaatcattag aacaacgcat tacgagtctt gagaatggtc 3480taaagccagt ttatgatatg gcaaaaacaa tctcctcatt gaacagggtt tgtgctgaga 3540tggttgcaaa atatgatctt ctggtgatga caaccggtcg ggcaacagca accgctgcgg 3600caactgaggc ttattgggcc gaacatggtc aaccaccacc tggaccatca ctttatgaag 3660aaagtgcgat tcggggtaag attgaatcta gagatgagac cgtccctcaa agtgttaggg 3720aggcattcaa caatctaaac agtaccactt cactaactga ggaaaatttt gggaaacctg 3780acatttcggc aaaggatttg agaaacatta tgtatgatca cttgcctggt tttggaactg 3840ctttccacca attagtacaa gtgatttgta aattgggaaa agatagcaac tcattggaca 3900tcattcatgc tgagttccag gccagcctgg ctgaaggaga ctctcctcaa tgtgccctaa 3960ttcaaattac aaaaagagtt ccaatcttcc aagatgctgc tccacctgtc atccacatcc 4020gctctcgagg tgacattccc cgagcttgcc agaaaagctt gcgtccagtc ccaccatcgc 4080ccaagattga tcgaggttgg gtatgtgttt ttcagcttca agatggtaaa acacttggac 4140tcaaaatttg agccaatctc ccttccctcc gaaagaggcg aataatagca gaggcttcaa 4200ctgctgaact atagggtacg ttacattaat gatacacttg tgagtatcag ccctggataa 4260tataagtcaa ttaaacgacc aagataaaat tgttcatatc tcgctagcag cttaaaatat 4320aaatgtaata ggagctatat ctctgacagt attataatca attgttatta agtaacccaa 4380accaaaagtg atgaagatta agaaaaacct acctcggctg agagagtgtt ttttcattaa 4440ccttcatctt gtaaacgttg agcaaaattg ttaaaaatat gaggcgggtt atattgccta 4500ctgctcctcc tgaatatatg gaggccatat accctgtcag gtcaaattca acaattgcta 4560gaggtggcaa cagcaataca ggcttcctga caccggagtc agtcaatggg gacactccat 4620cgaatccact caggccaatt gccgatgaca ccatcgacca tgccagccac acaccaggca 4680gtgtgtcatc agcattcatc cttgaagcta tggtgaatgt catatcgggc cccaaagtgc 4740taatgaagca aattccaatt tggcttcctc taggtgtcgc tgatcaaaag acctacagct 4800ttgactcaac tacggccgcc atcatgcttg cttcatacac tatcacccat ttcggcaagg 4860caaccaatcc acttgtcaga gtcaatcggc tgggtcctgg aatcccggat catcccctca 4920ggctcctgcg aattggaaac caggctttcc tccaggagtt cgttcttccg ccagtccaac 4980taccccagta tttcaccttt gatttgacag cactcaaact gatcacccaa ccactgcctg 5040ctgcaacatg gaccgatgac actccaacag gatcaaatgg agcgttgcgt ccaggaattt 5100catttcatcc aaaacttcgc cccattcttt tacccaacaa aagtgggaag aaggggaaca 5160gtgccgatct aacatctccg gagaaaatcc aagcaataat gacttcactc caggacttta 5220agatcgttcc aattgatcca accaaaaata tcatgggaat cgaagtgcca gaaactctgg 5280tccacaagct gaccggtaag aaggtgactt ctaaaaatgg acaaccaatc atccctgttc 5340ttttgccaaa gtacattggg ttggacccgg tggctccagg agacctcacc atggtaatca 5400cacaggattg tgacacgtgt cattctcctg caagtcttcc agctgtgatt gagaagtaat 5460tgcaataatt gactcagatc cagttttata gaatcttctc agggatagtg ataacatcta 5520tttagtaatc cgtccattag aggagacact tttaattgat caatatacta aaggtgcttt 5580acaccattgt cttttttctc tcctaaatgt agaacttaac aaaagactca taatatactt 5640gtttttaaag gattgattga tgaaagatca taactaataa cattacaaat aatcctacta 5700taatcaatac ggtgattcaa atgttaatct ttctcattgc acatactttt tgcccttatc 5760ctcaaattgc ctgcatgctt acatctgagg atagccagtg tgacttggat tggaaatgtg 5820gagaaaaaat cgggacccat ttctaggttg ttcacaatcc aagtacagac attgcccttc 5880taattaagaa aaaatcggcg atgaagatta agccgacagt gagcgtaatc ttcatctctc 5940ttagattatt tgttttccag agtaggggtc gtcaggtcct tttcaatcgt gtaaccaaaa 6000taaactccac tagaaggata ttgtggggca acaacacaat gggcgttaca ggaatattgc 6060agttacctcg tgatcgattc aagaggacat cattctttct ttgggtaatt atccttttcc 6120aaagaacatt ttccatccca cttggagtca tccacaatag cacattacag gttagtgatg 6180tcgacaaact agtttgtcgt gacaaactgt catccacaaa tcaattgaga tcagttggac 6240tgaatctcga agggaatgga gtggcaactg acgtgccatc tgcaactaaa agatggggct 6300tcaggtccgg tgtcccacca aaggtggtca attatgaagc tggtgaatgg gctgaaaact 6360gctacaatct tgaaatcaaa aaacctgacg ggagtgagtg tctaccagca gcgccagacg 6420ggattcgggg cttcccccgg tgccggtatg tgcacaaagt atcaggaacg ggaccgtgtg 6480ccggagactt tgccttccat aaagagggtg ctttcttcct gtatgatcga cttgcttcca 6540cagttatcta ccgaggaacg actttcgctg aaggtgtcgt tgcatttctg atactgcccc 6600aagctaagaa ggacttcttc agctcacacc ccttgagaga gccggtcaat gcaacggagg 6660acccgtctag tggctactat tctaccacaa ttagatatca ggctaccggt tttggaacca 6720atgagacaga gtacttgttc gaggttgaca atttgaccta cgtccaactt gaatcaagat 6780tcacaccaca gtttctgctc cagctgaatg agacaatata tacaagtggg aaaaggagca 6840ataccacggg aaaactaatt tggaaggtca accccgaaat tgatacaaca atcggggagt 6900gggccttctg ggaaactaaa aaaacctcac tagaaaaatt cgcagtgaag agttgtcttt 6960cacagttgta tcaaacggag ccaaaaacat cagtggtcag agtccggcgc gaacttcttc 7020cgacccaggg accaacacaa caactgaaga ccacaaaatc atggcttcag aaaattcctc 7080tgcaatggtt caagtgcaca gtcaaggaag ggaagctgca gtgtcgcatc taacaaccct 7140tgccacaatc tccacgagtc cccaatccct cacaaccaaa ccaggtccgg acaacagcac 7200ccataataca cccgtgtata aacttgacat ctctgaggca actcaagttg aacaacatca 7260ccgcagaaca gacaacgaca gcacagcctc cgacactccc tctgccacga ccgcagccgg 7320acccccaaaa gcagagaaca ccaacacgag caagagcact gacttcctgg accccgccac 7380cacaacaagt ccccaaaacc acagcgagac cgctggcaac aacaacactc atcaccaaga 7440taccggagaa gagagtgcca gcagcgggaa gctaggctta attaccaata ctattgctgg 7500agtcgcagga ctgatcacag gcgggagaag aactcgaaga gaagcaattg tcaatgctca 7560acccaaatgc aaccctaatt tacattactg gactactcag gatgaaggtg ctgcaatcgg 7620actggcctgg ataccatatt tcgggccagc agccgaggga atttacatag aggggctaat 7680gcacaatcaa gatggtttaa tctgtgggtt gagacagctg gccaacgaga cgactcaagc 7740tcttcaactg ttcctgagag ccacaactga gctacgcacc ttttcaatcc tcaaccgtaa 7800ggcaattgat ttcttgctgc agcgatgggg cggcacatgc cacattctgg gaccggactg 7860ctgtatcgaa ccacatgatt ggaccaagaa cataacagac aaaattgatc agattattca 7920tgattttgtt gataaaaccc ttccggacca gggggacaat gacaattggt ggacaggatg 7980gagacaatgg ataccggcag gtattggagt tacaggcgtt ataattgcag ttatcgcttt 8040attctgtata tgcaaatttg tcttttagtt tttcttcaga ttgcttcatg gaaaagctca 8100gcctcaaatc aatgaaacca ggatttaatt atatggatta cttgaatcta agattacttg 8160acaaatgata atataataca ctggagcttt aaacatagcc aatgtgattc taactccttt 8220aaactcacag ttaatcataa acaaggtttg acatcaatct agttatctct ttgagaatga 8280taaacttgat gaagattaag aaaaaggtaa tctttcgatt atctttaatc ttcatccttg 8340attctacaat catgacagtt gtctttagtg acaagggaaa gaagcctttt tattaagttg 8400taataatcag atctgcgaac cggtagagtt tagttgcaac ctaacacaca taaagcattg 8460gtcaaaaagt caatagaaat ttaaacagtg agtggagaca acttttaaat ggaagcttca 8520tatgagagag gacgcccacg agctgccaga cagcattcaa gggatggaca cgaccaccat 8580gttcgagcac gatcatcatc cagagagaat tatcgaggtg agtaccgtca atcaaggagc 8640gcctcacaag tgcgcgttcc tactgtattt cataagaaga gagttgaacc attaacagtt 8700cctccagcac ctaaagacat atgtccgacc ttgaaaaaag gatttttgtg tgacagtagt 8760ttttgcaaaa aagatcacca gttggagagt ttaactgata gggaattact cctactaatc 8820gcccgtaaga cttgtggatc agtagaacaa caattaaata taactgcacc caaggactcg 8880cgcttagcaa atccaacggc tgatgatttc cagcaagagg aaggtccaaa aattaccttg 8940ttgacactga tcaagacggc agaacactgg gcgagacaag acatcagaac catagaggat 9000tcaaaattaa gagcattgtt gactctatgt gctgtgatga cgaggaaatt ctcaaaatcc 9060cagctgagtc ttttatgtga gacacaccta aggcgcgagg ggcttgggca agatcaggca 9120gaacccgttc tcgaagtata tcaacgatta cacagtgata aaggaggcag ttttgaagct 9180gcactatggc aacaatggga ccgacaatcc ctaattatgt ttatcactgc attcttgaat 9240attgctctcc agttaccgtg tgaaagttct gctgtcgttg tttcagggtt aagaacattg 9300gttcctcaat cagataatga ggaagcttca accaacccgg ggacatgctc atggtctgat 9360gagggtaccc cttaataagg ctgactaaaa cactatataa ccttctactt gatcacaata 9420ctccgtatac ctatcatcat atatttaatc aagacgatat cctttaaaac ttattcagta 9480ctataatcac tctcgtttca aattaataag atgtgcatga ttgccctaat atatgaagag 9540gtatgataca accctaacag tgatcaaaga aaatcataat ctcgtatcgc tcgaaatata 9600acctgccaag catacctctt gcacaaagtg attcttgtac acaaataatg ttttactcta 9660caggaggtag caacgatcca tcccatcaaa aaataagtat ttcatgactt actaatgatc 9720tcttaaaata ttaagaaaaa ctgacggaac ataaattctt tatgcttcaa gctgtggagg 9780aggtgtttgg tattggctat tgttatatta caatcaataa caagcttgta aaaatattgt 9840tcttgtttca agaggtagat tgtgaccgga aatgctaaac taatgatgaa gattaatgcg 9900gaggtctgat aagaataaac cttattattc agattaggcc ccaagaggca ttcttcatct 9960ccttttagca aagtactatt tcagggtagt ccaattagtg gcacgtcttt tagctgtata 10020tcagtcgccc ctgagatacg ccacaaaagt gtctctaagc taaattggtc tgtacacatc 10080ccatacattg tattaggggc aataatatct aattgaactt agccgtttaa aatttagtgc 10140ataaatctgg gctaacacca ccaggtcaac tccattggct gaaaagaagc ttacctacaa 10200cgaacatcac tttgagcgcc ctcacaatta aaaaatagga acgtcgttcc aacaatcgag 10260cgcaaggttt caaggttgaa ctgagagtgt ctagacaaca aaatattgat actccagaca 10320ccaagcaaga cctgagaaaa aaccatggct aaagctacgg gacgatacaa tctaatatcg 10380cccaaaaagg acctggagaa aggggttgtc ttaagcgacc tctgtaactt cttagttagc 10440caaactattc aggggtggaa ggtttattgg gctggtattg agtttgatgt gactcacaaa 10500ggaatggccc tattgcatag actgaaaact aatgactttg cccctgcatg gtcaataaca 10560aggaatctct ttcctcattt atttcaaaat ccgaattcca caattgaatc accgctgtgg 10620gcattgagag tcatccttgc agcagggata caggaccagc tgattgacca gtctttgatt 10680gaacccttag caggagccct tggtctgatc tctgattggc tgctaacaac caacactaac 10740catttcaaca tgcgaacaca acgtgtcaag gaacaattga gcccaaaaat gctgtcgttg 10800attcgatcca atattctcaa gtttattaac aaattggatg ctctacatgt cgtgaactac 10860aacggattgt tgagcagtat tgaaattgga actcaaaatc atataatcat cataactcga 10920actaacatgg gttttctggt ggagctccaa gaacccgaca aatcggcaat gaaccgcatg 10980aagcctgggc cggcgaaatt ttccctcctt catgagtcca cactgaaagc atttacacaa 11040ggatcctcga cacgaatgca aagtttgatt cttgaattta atagctctct tgctatctaa 11100ctaaggtaga atacttcata ttgagctaac tcatatatgc tgactcaata gttatcttga 11160catctctgct ttcataatca gatatataag cataataaat aaatactcat atttcttgat 11220aatttgttta accacagata aatcctcact gtaagccagc ttccaagttg acacccttac 11280aaaaaccagg actcagaatc cctcaaacaa gagattccaa gacaacatca tagaattgct 11340ttattatatg aataagcatt ttatcaccag aaatcctata tactaaatgg ttaattgtaa 11400ctgaacccgc aggtcacatg tgttaggttt cacagattct atatattact aactctatac 11460tcgtaattaa cattagataa gtagattaag aaaaaagcct gaggaagatt aagaaaaact 11520gcttattggg tctttccgtg ttttagatga agcagttgaa attcttcctc ttgatattaa 11580atggctacac aacataccca atacccagac gctaggttat catcaccaat tgtattggac 11640caatgtgacc tagtcactag agcttgcggg ttatattcat catactccct taatccgcaa 11700ctacgcaact gtaaactccc gaaacatatc taccgtttga aatacgatgt aactgttacc 11760aagttcttga gtgatgtacc agtggcgaca ttgcccatag atttcatagt cccagttctt 11820ctcaaggcac tgtcaggcaa tggattctgt cctgttgagc cgcggtgcca acagttctta 11880gatgaaatca ttaagtacac aatgcaagat gctctcttct tgaaatatta tctcaaaaat 11940gtgggtgctc aagaagactg tgttgatgaa cactttcaag agaaaatctt atcttcaatt 12000cagggcaatg aatttttaca tcaaatgttt ttctggtatg atctggctat tttaactcga 12060aggggtagat taaatcgagg aaactctaga tcaacatggt ttgttcatga tgatttaata 12120gacatcttag gctatgggga ctatgttttt tggaagatcc caatttcaat gttaccactg 12180aacacacaag gaatccccca tgctgctatg gactggtatc aggcatcagt attcaaagaa 12240gcggttcaag ggcatacaca cattgtttct gtttctactg ccgacgtctt gataatgtgc 12300aaagatttaa ttacatgtcg attcaacaca actctaatct caaaaatagc agagattgag 12360gatccagttt gttctgatta tcccaatttt aagattgtgt ctatgcttta ccagagcgga 12420gattacttac tctccatatt agggtctgat gggtataaaa ttattaagtt cctcgaacca 12480ttgtgcttgg ccaaaattca attatgctca aagtacactg agaggaaggg ccgattctta 12540acacaaatgc atttagctgt aaatcacacc ctagaagaaa ttacagaaat gcgtgcacta 12600aagccttcac aggctcaaaa gatccgtgaa ttccatagaa cattgataag gctggagatg 12660acgccacaac aactttgtga gctattttcc attcaaaaac actgggggca tcctgtgcta 12720catagtgaaa cagcaatcca aaaagttaaa aaacatgcta cggtgctaaa agcattacgc 12780cctatagtga ttttcgagac atactgtgtt tttaaatata gtattgccaa acattatttt 12840gatagtcaag gatcttggta cagtgttact tcagatagga atctaacacc gggtcttaat 12900tcttatatca aaagaaatca attccctccg ttgccaatga ttaaagaact actatgggaa 12960ttttaccacc ttgaccaccc tccacttttc tcaaccaaaa ttattagtga cttaagtatt 13020tttataaaag acagagctac cgcagtagaa aggacatgct gggatgcagt attcgagcct 13080aatgttctag gatataatcc acctcacaaa tttagtacta aacgtgtacc ggaacaattt 13140ttagagcaag aaaacttttc tattgagaat gttctttcct acgcacaaaa actcgagtat 13200ctactaccac aatatcggaa cttttctttc tcattgaaag agaaagagtt gaatgtaggt 13260agaaccttcg gaaaattgcc ttatccgact cgcaatgttc aaacactttg tgaagctctg 13320ttagctgatg gtcttgctaa agcatttcct agcaatatga tggtagttac ggaacgtgag 13380caaaaagaaa gcttattgca tcaagcatca tggcaccaca caagtgatga ttttggtgaa 13440catgccacag ttagagggag tagctttgta actgatttag agaaatacaa tcttgcattt 13500agatatgagt ttacagcacc ttttatagaa tattgcaacc gttgctatgg tgttaagaat 13560gtttttaatt ggatgcatta tacaatccca cagtgttata tgcatgtcag tgattattat 13620aatccaccac ataacctcac actggagaat cgagacaacc cccccgaagg gcctagttca 13680tacaggggtc atatgggagg gattgaagga ctgcaacaaa aactctggac aagtatttca 13740tgtgctcaaa tttctttagt tgaaattaag actggtttta agttacgctc agctgtgatg 13800ggtgacaatc agtgcattac tgttttatca gtcttcccct tagagactga cgcagacgag 13860caggaacaga gcgccgaaga caatgcagcg agggtggccg ccagcctagc aaaagttaca 13920agtgcctgtg gaatcttttt aaaacctgat gaaacatttg tacattcagg ttttatctat 13980tttggaaaaa aacaatattt gaatggggtc caattgcctc agtcccttaa aacggctgca 14040agaatggcac cattgtctga tgcaattttt gatgatcttc aagggaccct ggctagtata 14100ggcactgctt ttgagcgatc catctctgag acacgacata tctttccttg caggataacc 14160gcagctttcc atacgttttt ttcggtgaga atcttgcaat atcatcatct cgggttcaat 14220aaaggttttg accttggaca gttaacactc ggcaaacctc tggatttcgg aacaatatca 14280ttggcactag cggtaccgca ggtgcttgga gggttatcct tcttgaatcc tgagaaatgt 14340ttctaccgga atctaggaga tccagttacc tcaggcttat tccagttaaa aacttatctc 14400cgaatgattg agatggatga tttattctta cctttaattg cgaagaaccc tgggaactgc 14460actgccattg actttgtgct aaatcctagc ggattaaatg tccctgggtc gcaagactta 14520acttcatttc tgcgccagat tgtacgcagg accatcaccc taagtgcgaa aaacaaactt 14580attaatacct tatttcatgc gtcagctgac ttcgaagacg aaatggtttg taaatggcta 14640ttatcatcaa ctcctgttat gagtcgtttt gcggccgata tcttttcacg cacgccgagc 14700gggaagcgat tgcaaattct aggatacctg gaaggaacac gcacattatt agcctctaag 14760atcatcaaca ataatacaga gacaccggtt ttggacagac tgaggaaaat aacattgcaa 14820aggtggagcc tatggtttag ttatcttgat cattgtgata atatcctggc ggaggcttta 14880acccaaataa cttgcacagt tgatttagca cagattctga gggaatattc atgggctcat 14940attttagagg gaagacctct tattggagcc acactcccat gtatgattga gcaattcaaa 15000gtgttttggc tgaaacccta cgaacaatgt ccgcagtgtt caaatgcaaa gcaaccaggt 15060gggaaaccat tcgtgtcagt ggcagtcaag aaacatattg ttagtgcatg gccgaacgca 15120tcccgaataa gctggactat cggggatgga atcccataca ttggatcaag gacagaagat 15180aagataggac aacctgctat taaaccaaaa tgtccttccg cagccttaag agaggccatt 15240gaattggcgt cccgtttaac atgggtaact caaggcagtt cgaacagtga cttgctaata 15300aaaccatttt tggaagcacg agtaaattta agtgttcaag aaatacttca aatgacccct 15360tcacattact caggaaatat tgttcacagg tacaacgatc aatacagtcc tcattctttc 15420atggccaatc gtatgagtaa ttcagcaacg cgattgattg tttctacaaa cactttaggt 15480gagttttcag gaggtggcca gtctgcacgc gacagcaata ttattttcca gaatgttata 15540aattatgcag ttgcactgtt cgatattaaa tttagaaaca ctgaggctac agatatccaa 15600tataatcgtg ctcaccttca tctaactaag tgttgcaccc gggaagtacc agctcagtat 15660ttaacataca catctacatt ggatttagat ttaacaagat accgagaaaa cgaattgatt 15720tatgacagta atcctctaaa aggaggactc aattgcaata tctcattcga taatccattt 15780ttccaaggta aacggctgaa cattatagaa gatgatctta ttcgactgcc tcacttatct 15840ggatgggagc tagccaagac catcatgcaa tcaattattt cagatagcaa caattcatct 15900acagacccaa ttagcagtgg agaaacaaga tcattcacta cccatttctt aacttatccc 15960aagataggac ttctgtacag ttttggggcc tttgtaagtt attatcttgg caatacaatt 16020cttcggacta agaaattaac acttgacaat tttttatatt acttaactac tcaaattcat 16080aatctaccac atcgctcatt gcgaatactt aagccaacat tcaaacatgc aagcgttatg

16140tcacggttaa tgagtattga tcctcatttt tctatttaca taggcggtgc tgcaggtgac 16200agaggactct cagatgcggc caggttattt ttgagaacgt ccatttcatc ttttcttaca 16260tttgtaaaag aatggataat taatcgcgga acaattgtcc ctttatggat agtatatccg 16320ctagagggtc aaaacccaac acctgtgaat aattttctct atcagatcgt agaactgctg 16380gtgcatgatt catcaagaca acaggctttt aaaactacca taagtgatca tgtacatcct 16440cacgacaatc ttgtttacac atgtaagagt acagccagca atttcttcca tgcatcattg 16500gcgtactgga ggagcagaca cagaaacagc aaccgaaaat acttggcaag agactcttca 16560actggatcaa gcacaaacaa cagtgatggt catattgaga gaagtcaaga acaaaccacc 16620agagatccac atgatggcac tgaacggaat ctagtcctac aaatgagcca tgaaataaaa 16680agaacgacaa ttccacaaga aaacacgcac cagggtccgt cgttccagtc ctttctaagt 16740gactctgctt gtggtacagc aaatccaaaa ctaaatttcg atcgatcgag acacaatgtg 16800aaatttcagg atcataactc ggcatccaag agggaaggtc atcaaataat ctcacaccgt 16860ctagtcctac ctttctttac attatctcaa gggacacgcc aattaacgtc atccaatgag 16920tcacaaaccc aagacgagat atcaaagtac ttacggcaat tgagatccgt cattgatacc 16980acagtttatt gtagatttac cggtatagtc tcgtccatgc attacaaact tgatgaggtc 17040ctttgggaaa tagagagttt caagtcggct gtgacgctag cagagggaga aggtgctggt 17100gccttactat tgattcagaa ataccaagtt aagaccttat ttttcaacac gctagctact 17160gagtccagta tagagtcaga aatagtatca ggaatgacta ctcctaggat gcttctacct 17220gttatgtcaa aattccataa tgaccaaatt gagattattc ttaacaactc agcaagccaa 17280ataacagaca taacaaatcc tacttggttt aaagaccaaa gagcaaggct acctaagcaa 17340gtcgaggtta taaccatgga tgcagagaca acagagaata taaacagatc gaaattgtac 17400gaagctgtat ataaattgat cttacaccat attgatccta gcgtattgaa agcagtggtc 17460cttaaagtct ttctaagtga tactgagggt atgttatggc taaatgataa tttagccccg 17520ttttttgcca ctggttattt aattaagcca ataacgtcaa gtgctagatc tagtgagtgg 17580tatctttgtc tgacgaactt cttatcaact acacgtaaga tgccacacca aaaccatctc 17640agttgtaaac aggtaatact tacggcattg caactgcaaa ttcaacgaag cccatactgg 17700ctaagtcatt taactcagta tgctgactgt gagttacatt taagttatat ccgccttggt 17760tttccatcat tagagaaagt actataccac aggtataacc tcgtcgattc aaaaagaggt 17820ccactagtct ctatcactca gcacttagca catcttagag cagagattcg agaattaact 17880aatgattata atcaacagcg acaaagtcgg actcaaacat atcactttat tcgtactgca 17940aaaggacgaa tcacaaaact agtcaatgat tatttaaaat tctttcttat tgtgcaagca 18000ttaaaacata atgggacatg gcaagctgag tttaagaaat taccagagtt gattagtgtg 18060tgcaataggt tctaccatat tagagattgc aattgtgaag aacgtttctt agttcaaacc 18120ttatatttac atagaatgca ggattctgaa gttaagctta tcgaaaggct gacagggctt 18180ctgagtttat ttccggatgg tctctacagg tttgattgaa ttaccgtgca tagtatcctg 18240atacttgcaa aggttggtta ttaacataca gattataaaa aactcataaa ttgctctcat 18300acatcatatt gatctaatct caataaacaa ctatttaaat aacgaaagga gtccctatat 18360tatatactat atttagcctc tctccctgcg tgataatcaa aaaattcaca atgcagcatg 18420tgtgacatat tactgccgca atgaatttaa cgcaacataa taaactctgc actctttata 18480attaagcttt aacgaaaggt ctgggctcat attgttattg atataataat gttgtatcaa 18540tatcctgtca gatggaatag tgttttggtt gataacacaa cttcttaaaa caaaattgat 18600ctttaagatt aagtttttta taattatcat tactttaatt tgtcgtttta aaaacggtga 18660tagccttaat ctttgtgtaa aataagagat taggtgtaat aaccttaaca tttttgtcta 18720gtaagctact atttcataca gaatgataaa attaaaagaa aaggcaggac tgtaaaatca 18780gaaatacctt ctttacaata tagcagacta gataataatc ttcgtgttaa tgataattaa 18840gacattgacc acgctcatca gaaggctcgc cagaataaac gttgcaaaaa ggattcctgg 18900aaaaatggtc gcacacaaaa atttaaaaat aaatctattt cttctttttt gtgtgtcca 1895930288PRTZaire ebolavirus 30Met Glu Ala Ser Tyr Glu Arg Gly Arg Pro Arg Ala Ala Arg Gln His1 5 10 15 Ser Arg Asp Gly His Asp His His Val Arg Ala Arg Ser Ser Ser Arg 20 25 30 Glu Asn Tyr Arg Gly Glu Tyr Arg Gln Ser Arg Ser Ala Ser Gln Val 35 40 45 Arg Val Pro Thr Val Phe His Lys Lys Arg Val Glu Pro Leu Thr Val 50 55 60 Pro Pro Ala Pro Lys Asp Ile Cys Pro Thr Leu Lys Lys Gly Phe Leu65 70 75 80 Cys Asp Ser Ser Phe Cys Lys Lys Asp His Gln Leu Glu Ser Leu Thr 85 90 95 Asp Arg Glu Leu Leu Leu Leu Ile Ala Arg Lys Thr Cys Gly Ser Val 100 105 110 Glu Gln Gln Leu Asn Ile Thr Ala Pro Lys Asp Ser Arg Leu Ala Asn 115 120 125 Pro Thr Ala Asp Asp Phe Gln Gln Glu Glu Gly Pro Lys Ile Thr Leu 130 135 140 Leu Thr Leu Ile Lys Thr Ala Glu His Trp Ala Arg Gln Asp Ile Arg145 150 155 160 Thr Ile Glu Asp Ser Lys Leu Arg Ala Leu Leu Thr Leu Cys Ala Val 165 170 175 Met Thr Arg Lys Phe Ser Lys Ser Gln Leu Ser Leu Leu Cys Glu Thr 180 185 190 His Leu Arg Arg Glu Gly Leu Gly Gln Asp Gln Ala Glu Pro Val Leu 195 200 205 Glu Val Tyr Gln Arg Leu His Ser Asp Lys Gly Gly Ser Phe Glu Ala 210 215 220 Ala Leu Trp Gln Gln Trp Asp Arg Gln Ser Leu Ile Met Phe Ile Thr225 230 235 240 Ala Phe Leu Asn Ile Ala Leu Gln Leu Pro Cys Glu Ser Ser Ala Val 245 250 255 Val Val Ser Gly Leu Arg Thr Leu Val Pro Gln Ser Asp Asn Glu Glu 260 265 270 Ala Ser Thr Asn Pro Gly Thr Cys Ser Trp Ser Asp Glu Gly Thr Pro 275 280 285 3110PRTZaire ebolavirus 31Ser Phe Lys Ala Ala Leu Ser Ser Leu Ala1 5 10 3210PRTZaire ebolavirus 32Tyr Phe Thr Phe Asp Leu Thr Ala Leu Lys1 5 10 339PRTZaire ebolavirus 33Leu Tyr Asp Arg Leu Ala Ser Thr Val1 5

Claims

1. A method to identify one or more agents that inhibit Ebola virus infection, comprising: a) contacting a host cell, one or more agents and recombinant infectious, biologically contained Ebola virus, the genome of which contains a deletion of sequences corresponding to Ebola virus VP30 sequences, which deletion is effective to prevent expression of functional VP30 upon infection of a cell with the recombinant virus; and b) identifying one or more agents that inhibit viral infection without substantially decreasing host cell viability.

2. The method of claim 1 wherein the host cell is contacted with the virus before or after the one or more agents.

3. The method of claim 1 wherein the host cell is contacted with the virus and the agents simultaneously.

4. The method of claim 1 wherein the identified agent inhibits the infectivity of the virus by at least 90%.

5. The method of claim 1 wherein the detected agent has an IC.sub.50 of less than about 10.0 .mu.M or a CC.sub.50 of more than about 0.1 .mu.M.

6. (canceled)

7. A method to identify inhibitors of filovirus glycoprotein receptor binding or fusion, comprising: a) contacting a host cell with one or more agents and a recombinant replication incompetent pseudotyped rhabdovirus comprising filovirus glycoprotein and a mutant negative sense rhabdovirus genome which lacks sequences for a rhabdovirus glycoprotein but comprises a sequence for a reporter protein; and b) identifying at least one agent that inhibits reporter protein levels or expression in the host cell.

8. The method of claim 7 wherein the rhabdovirus is VSV.

9. The method of claim 7 wherein the filovirus glycoprotein is a Marburg virus glycoprotein or an Ebolavirus glycoprotein.

10. (canceled)

11. The method of claim 7 wherein the reporter protein levels or expression in the presence of the one or more agents is compared to reporter protein levels or expression in the absence of the agent(s).

12. The method of claim 7 wherein at least one agent inhibits filovirus glycoprotein binding.

13. The method of claim 7 wherein at least one agent inhibits filovirus glycoprotein fusion.

14. (canceled)

15. The method of claim 7 wherein the host cell is a mammalian cell.

16. (canceled)

17. The method of claim 1 wherein the at least one agent contacted with the cell is a triphenylethylene, an inhibitor of calcium-independent phospholipase A.sub.2 and/or of magnesium-dependent phosphatidate phosphohydrolase, an inhibitor of PGE.sub.2 synthase, a steroid, dopamine antagonist, anticholinergic or an Hsp90 inhibitor.

18. The method of claim 1 wherein the at least one agent is a compound of formula (I)-(XIII).

19-21. (canceled)

22. A method to prevent, inhibit or treat filovirus infection in a mammal, comprising administering to the mammal an effective amount of a compound of formula (I)-(XIII) or an effective amount of a pharmaceutical composition comprising an agent, wherein the composition comprises a calcium channel blocker, a tetranortriterpenoid, a sigma receptor agonist, triphenylethylene, an inhibitor of calcium-independent phospholipase A.sub.2, an inhibitor of magnesium-dependent phosphatidate phosphohydrolase, an inhibitor of PGE.sub.2 synthase, a steroid, adopamine antagonist, an inhibitor of Hsp90, or an anticholinergic.

23. (canceled)

24. The method of claim 22 wherein the mammal is a human.

25. The method of claim 22 wherein the agent is bepridil hydrochloride, clomiphene citrate, benzotropin mesylate, 7-deacetoxy-3-deacetyl-7-oxokhivorin, 1,2alpha-epoxy-7-deacetoxy-7-oxodihxdrogedunin, epoxygedunin, 1,3-dideacetyl-7-deacetoxy-7-oxokhivorin, gedunin, tamoxifen citrate, fluspirilene, raloxifene hydrochloride, bromoenol lactone, cortexolone maleate, (R,R)-cis,diethyl tetrahydro-2,8-chrysenediol, MK-866, L-687, 384 hydrochloride, cycloheximide, gedunol, dihydrogedunin, 3beta-acetoxydeoxodihydrogedunin, 3alpha-hydroxydeoxodihydrogedunin, deacetoxy-7-oxogedunin, 3beta-hydroxydeoxodihydrogedunin, deacetoxy-7-oxogedunin, 1,2alpha-epoxydeacetoxydihydrogedunin, 3beta-hydroxydeoxydesacetoxy-7-oxogedunin, tridesacetoxykhivorin, 1,3-dideacetylkhivorin, heudelottin C, geldanamycin, 17-allylamino-17-demethoxygeldanamycin, 4-[4-(2,3-dihydro-1,4-benzodioxin-6-yl)-5-methyl-1H-pyrazol-3-yl]-6-ethyl- -1,3-benzenediol, 6-phenylimidazo[2,1-b]-1,3,4-thiadiazole-2-sulfonamide, geldanamycin, 17-Allylamino-17-demethoxygeldanamycin, 4-[4-(2,3-Dihydro-1,4-benzodioxin-6-yl)-5-methyl-1H-pyrazol-3-yl]-6-ethyl- -1,3-benzenediol, 6-Phenylimidazo[2,1-b]-1,3,4-thiadiazole-2-sulfonamide, or any combination thereof.

26. The method of claim 22 wherein the agent is orally, intravenously or subcutaneously administered.

27-30. (canceled)

31. The method of claim 7 wherein the at least one agent contacted with the cell is a triphenylethylene, an inhibitor of calcium-independent phospholipase A.sub.2 and/or of magnesium-dependent phosphatidate phosphohydrolase, an inhibitor of PGE.sub.2 synthase, a steroid, dopamine antagonist, anticholinergic or an Hsp90 inhibitor.

32. The method of claim 7 wherein the at least one agent is a compound of formula (I)-(XII).