Patent Publication Number: US-2022233681-A1

Title: Live attenuated oral vaccine against covid-19 and typhoid fever

Description:
CROSS REFERENCE TO RELATED APPLICATIONS 
     The present application claims priority benefit of U.S. Provisional Application No. 63/133,131, filed Dec. 31, 2020, which is hereby incorporated by reference in its entirety. 
    
    
     REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY VIA EFS-WEB 
     The content of the electronically submitted sequence listing in ASCII text file format (Name: 4661_0010001_Seqlisting_ST25.txt; Size: 158,919 Bytes; and Date of Creation: Dec. 22, 2021) is hereby incorporated by reference in its entirety. 
     FIELD OF THE DISCLOSURE 
     The present disclosure is generally related to bioengineering and provides multivalent vaccines for protection against COVID-19 and/or typhoid fever. 
     BACKGROUND OF THE DISCLOSURE 
     Severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2), the causative agent of COVID-19, is spreading globally in the most significant pandemic of the past 100 years (World Health Organization,  WHO Director - General&#39;s opening remarks at the media briefing on COVID -19-11 (March 2020); World Health Organaization,  Timeline of WHO&#39;s response to COVID -19. 29 Jun. 2020, https://www.who.int/news-room/detail/29-06-2020-covidtimeline). As of 27 Dec. 2020, there were 79,232,555 million cases and 1,754,493 deaths worldwide (GISAID,  Coronavirus COVID -19  Global Cases by Johns Hopkins CSSE , https://www.gisaid.org/epiflu-applications/global-cases-covid-19/ (2020)). As of the same time, an approved vaccine was urgently needed. As of December 2020, there were about 236 COVID-19 vaccine approaches under development (FastCures, The Milken Institute,  COVID -19  treatment and vaccine tracker  (2020), https://covid-19tracker.milkeninstitute.org/). The most advanced approach (using mRNA technologies) resulted in 2 successful candidates being rolled out to the public (https://www.fda.gov/emergency-preparedness-and-response/coronavirus-disease-2019-covid-19/pfizer-biontech-covid-19-vaccine; https://www.fda.gov/emergency-preparedness-and-response/coronavirus-disease-2019-covid-19/moderna-covid-19-vaccine). 
     Other candidates include recombinant chimpanzee adenovirus, nucleic acid (mRNA and DNA), and protein/adjuvant vaccines, all expressing or containing the spike (S) glycoprotein of SARS-CoV-2. Many approaches suffer from the downstream challenges to scale up manufacturing of these vaccines (and adjuvant) and implement widespread vaccination in a safe and efficient manner. However, all these candidates likely will require storage and shipment at &lt;−60° C., and widespread distribution at these temperatures will require dry ice for shipping and ultra-low freezers for storage, which in turn are dependent on a reliable electrical source. These requirements are problematic for many of the health care facilities that will be administering the vaccine, including physician&#39;s offices and pharmacies all over the world, as such facilities rarely have ultra-low freezers. In addition, in low and middle-income countries, electricity is minimal, unreliable and/or absent in many areas and populations, creating a significant challenge for vaccine distribution. Finally, given that COVID-19 is highly infectious and easily spread, requiring populations to congregate at hospitals, clinics or vaccine administration centers to receive vaccination (likely requiring multiple visits) will pose a confounding problem. 
       Salmonella Typhi  Ty21a typhoid vaccine (Vivotif®) (Kopecko, D. J., et al.,  Int J Med Microbiol,  299(4):233-46 (2009)) is one of only a few live, oral, attenuated bacterial vaccines licensed in the US. Ty21a, when administered for a one week period, affords sustained protection from typhoid fever for 7 years with efficacies ranging from 62-96% as reported in Chilean/Egyptian field trials (Levine, M. M.  Typhoid fever vaccines . In: Plotkin, S. A., Orenstein, W. A., eds.  Vaccines,  3 rd  ed. Philadelphia, Pa., WB Saunders Company, 1999:781-814; Levine, M. M., et al.,  Vaccine  17 Suppl 2:S22-7 (1999); Wandan, M. H., et al.,  J Infect Dis.  145(3):292-5 (1982)), and it has had an unrivaled safety record during the past 25 years (Gilman, R. H., et al.,  J Infect Dis.  136(6):717-23 (1977); Cryz, S. J., Jr.,  Lancet  341(8836):49-50 (1993); Simanjuntak, C. H., et al.,  Lancet  338(8774):1055-9 (1991); Levine, M. M., et al.,  The efficacy of attenuated Salmonella typhi oral vaccine strain Ty 21 a evaluated in controlled field trials . In: J. Holmgren, A. Lindberg, and R. Mollby, eds.  Development of vaccines against diarrhea.  11th Nobel Conference, Stockholm, 1985. Studentliteratur, Lund, Sweden, 1986:90-101). As of 2013, there has never been a reported case of bacteremic dissemination of Ty21a after administration to more than 150 million recipients (Kopecko, D. J., et al.,  Int J Med Microbiol.  299(4):233-46 (2009)), and Ty21a is nonpathogenic even when given at 100 times the standard dose (Levine, M. M., et al.,  Vaccine  17 Suppl 2:S22-7 (1999)). Also, there are no reports of post-vaccination inflammatory arthritis (e.g., Reiter&#39;s syndrome) with Ty21a, a potential problem with other live attenuated vectors including non-typhoid  Salmonella, Shigella , and  Yersinia . Ty21a can be foam-dried, which provides for temperature stabilization (Ohtake, S., et al.,  Vaccine  29(15): p. 2761-71 (2011)). 
     Ty21a has been used for expression of heterologous antigens from plasmids (U.S. Pat. Nos. 7,541,043, 8,071,084, 8,337,832, and 8,992,943; Chin&#39;ombe, N.,  Viruses  5(9):2062-78 (2013); and Frey, S. E., et al.,  Vaccine  31(42):4874-80 (2013)). Additionally, recombinant genome-integrated Ty21a strains have been used to stably express the 0-antigens of  Shigella sonnei  (U.S. Pat. Nos. 9,750,793 and 10,695,415), and Ty21a has been used to express the major virulence factors of enterotoxigenic  Escherichia coli  (ETEC) (Wai, T. et al.,  Amer Soc Trop Med  &amp;  Hygiene  101:539 (2019); U.S. Publication No. 2020/0376107 A1) and the protective antigen (PA) of  B. anthracis  (Sim, B. K. L., et al.,  NPJ Vaccines  2:17 (2017). A recent study described the beneficial nonspecific effects of oral vaccination with live-attenuated  Salmonella Typhi  strain Ty21a through the innate immune system (Pennington, S. H., et al.,  Sci Adv.  5(2):eaau6849 (2019)). 
     To date, there is no reported use of live attenuated Ty21a for expression of a heterologous viral antigen, nor use of such a vector to treat, prevent, or reduce the incidence of a viral infection. 
     SUMMARY OF THE DISCLOSURE 
     The present disclosure relates generally to the development of bivalent or multivalent, live, attenuated compositions (e.g., vaccine compositions) for protection against COVID-19 caused by SARS-CoV-2. Disclosed herein are constructed vaccine strains of  Salmonella typhi  in which one or more SARS-CoV-2-specific antigenic sequences from the spike (S), membrane (M), and/or nucleocapsid (N) proteins are recombineered into the chromosome of  Salmonella typhi  Ty21a such that the one or more antigenic sequences are stably expressed. The resulting recombineered Ty21a-SARS-CoV-2 strains can be used, for example, in orally or intranasally administered vaccine preparations to elicit an immune response to said antigens and to treat, prevent, or reduce the incidence of COVID-19 in an individual. In some aspects, the recombineered Ty21a-SARS-CoV-2 strains of the present disclosure are constructed to co-express the concerted YbaS-GadBC acid resistance (AR) system (SEQ ID NO: 20) and the one or more SARS-CoV-2-specific antigenic sequences. 
     Certain aspects of the present disclosure are directed to a transgenic  Salmonella typhi  Ty21a comprising a chromosome with one or more heterologous nucleic acid regions, wherein the heterologous nucleic acid regions encode one or more SARS-CoV-2 viral antigens and are integrated into the  Salmonella typhi  Ty21a chromosome, and wherein the transgenic  Salmonella typhi  Ty21a stably expresses the one or more SARS-CoV-2 viral antigens. 
     In some aspects, the heterologous nucleic acid regions comprise: (a) a nucleic acid sequence encoding a SARS-CoV-2 spike (S) protein or one or more antigenic fragments thereof, (b) a nucleic acid sequence encoding one or more antigens retaining at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% amino acid residue homology to a native SARS-CoV-2 S protein antigen, (c) a nucleic acid sequence encoding a SARS-CoV-2 S protein ectodomain (eS) or one or more antigenic fragments thereof, (d) a nucleic acid encoding one or more antigens retaining at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% amino acid residue homology to a native SARS-CoV-2 eS antigen, (e) a nucleic acid sequence encoding a SARS-CoV2 Membrane (M) protein or one or more antigenic fragments thereof, (f) a nucleic acid sequence encoding one or more antigens retaining at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% amino acid residue homology to a native SARS-CoV-2 M protein antigen, (g) a nucleic acid sequence encoding a SARS-CoV2 Nucleocapsid (N) protein or one or more antigenic fragments thereof, (h) a nucleic acid sequence encoding one or more antigens retaining at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% amino acid residue homology to a native SARS-CoV-2 N protein antigen, or (i) any combination thereof. 
     In some aspects, the heterologous nucleic acid regions encode a full length SARS-CoV-2 M protein. In some aspects, the full length SARS-CoV-2 M protein comprises the amino acid sequence of SEQ ID NO: 2. 
     In some aspects, the heterologous nucleic acid regions encode a full length SARS-CoV-2 N protein. In some aspects, the full length SARS-CoV-2 M protein comprises the amino acid sequence of SEQ ID NO: 3. 
     In some aspects, the heterologous nucleic acid regions encode a modified SARS-CoV-2 S protein Receptor Binding Domain (RBD) or a modified SARS-CoV-2 eS. In some aspects, the modified SARS-CoV-2 S protein RBD or the modified SARS-CoV-2 eS comprises a D614G substitution. In some aspects, the modified SARS-CoV-2 S protein RBD or the modified SARS-CoV-2 eS comprises a GSAS substitution at amino acid residues 682-685. In some aspects, the modified SARS-CoV-2 S protein RBD or the modified SARS-CoV-2 eS comprises proline substitutions at amino acid residues 986 and 987. In some aspects, the modified SARS-CoV-2 S protein RBD or the modified SARS-CoV-2 eS comprises a C-terminal T4 fibritin trimerization motif (SEQ ID NO: 12). In some aspects, the modified SARS-CoV-2 S protein RBD or the modified SARS-CoV-2 eS comprises F817P, A892P, A899P, and A942P substitutions. In some aspects, the modified SARS-CoV-2 RBD comprises the amino acid sequence of SEQ ID NO: 11. In some aspects, the modified SARS-CoV-2-eS comprises a hemolysin A (HLYA) signal sequence (SEQ ID NO: 13). In some aspects, the modified SARS-CoV-2 eS comprises the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 5. In some aspects, the heterologous nucleic acid regions encoding the modified SARS-CoV-2 eS comprise a nucleic acid sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90% at least 95%, at least 99%, or 100% identical to the nucleic acid sequence of SEQ ID NO: 6. 
     In some aspects, the heterologous nucleic acid regions encoding the modified SARS-CoV-2 S protein RBD or the modified SARS-CoV-2 eS further encode a full length SARS-CoV2 M protein and a full length SARS-CoV2 N protein. In some aspects, the full length SARS-CoV-2 M protein comprises the amino acid sequence of SEQ ID NO: 2, and the full length SARS-CoV-2 N protein comprises the amino acid sequence of SEQ ID NO: 3. 
     In some aspects, the heterologous nucleic acid regions encode a full length SARS-CoV-2 M protein and a full length SARS-CoV-2 N protein. In some aspects, the full length SARS-CoV-2 M protein comprises the amino acid sequence of SEQ ID NO: 2, and the full length SARS-CoV-2 N protein comprises the amino acid sequence of SEQ ID NO: 3. In some aspects, the heterologous nucleic acid regions encoding the full length SARS-CoV-2 M protein and the full length SARS-CoV-2 N protein comprise a nucleic acid sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90% at least 95%, at least 99%, or 100% identical to the nucleic acid sequence of SEQ ID NO: 7. 
     In some aspects, the transgenic  Salmonella typhi  Ty21a further comprises a heterologous acid resistance gene cassette integrated into the  Salmonella typhi  Ty21a chromosome. In some aspects, the heterologous acid resistance gene cassette comprises a gene selected from the group consisting of: a YbaS gene (SEQ ID NO: 15) or a fragment thereof, a GadB gene (SEQ ID NO: 17) or fragment thereof, a GadC gene (SEQ ID NO: 19) or fragment thereof, a GadA gene or fragment thereof (SEQ ID NO: 22), and any combination thereof. In some aspects, the heterologous acid resistance gene cassette comprises a sequence having at least 80%, 85%, 90%, 95%, or 100% sequence identity to one or more nucleic acid sequences selected from: SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 20, and SEQ ID NO: 22. In some aspects, the heterologous nucleic acid regions of the transgenic  Salmonella typhi  Ty21a comprising the heterologous acid resistance gene cassette comprise a nucleic acid sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90% at least 95%, at least 99%, or 100% identical to the nucleic acid sequence of SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10. In some aspects, the transgenic  Salmonella typhi  Ty21a comprising the heterologous acid resistance gene cassette is more acid stable at pH 2.5 than  Salmonella typhi  Ty21a without the integrated acid resistance gene. 
     Certain aspects of the present disclosure are directed to a composition, e.g., a bacterial composition, comprising a transgenic  Salmonella typhi  Ty21a disclosed herein in combination with a carrier, e.g., such that the carrier renders the composition suitable for pharmaceutical use. 
     Certain aspects of the present disclosure are directed to a vaccine comprising a composition disclosed herein. Certain aspects of the present disclosure are directed to a vaccine comprising a transgenic  Salmonella typhi  Ty21a disclosed herein in combination with a carrier, e.g., such that the carrier renders the vaccine suitable for pharmaceutical use. In some aspects, the vaccine is suitable for oral or inhaled administration. 
     In some aspects, the vaccine is protective against COVID-19. In some aspects, the vaccine is protective against typhoid fever. In some aspects, the vaccine is protective against COVID-19 and typhoid fever. 
     In some aspects, the vaccine is foam dried. In some aspects, the vaccine is foam dried, rendering the vaccine at least 90% stable, at least 95% stable, at least 99% stable, or 100% stable at room temperature for at least one month. In some aspects, the vaccine is foam dried, rendering the vaccine at least 40% stable, at least 50% stable, at least 60% stable, at least 70% stable, at least 80% stable, at least 90% stable, at least 95% stable, at least 99% stable, or 100% stable at room temperature for at least one year. In some aspects, the vaccine is foam dried, rendering the vaccine at least 40% stable, at least 50% stable, at least 60% stable, at least 70% stable, at least 80% stable, at least 90% stable, at least 95% stable, at least 99% stable, or 100% stable at room temperature for at least two years. In some aspects, the vaccine is foam dried and has a shelf-life of at least 3 months, at least 6 months, at least 9 months, at least 1 year, at least 2 years, at least 3 years, at least 4 years, at least 5 years at 4° C. In some aspects, the vaccine is foam dried and has a shelf-life of 6-24 months, 1-2 years, 1-5 years, 1-10 years, or 5-10 years at 4° C. 
     In some aspects, administration of a vaccine or composition of the present disclosure to one or more individuals or a human population reduces the incidence of COVID-19 in those individuals or that human population subsequently exposed to pathogenic SARS-CoV-2. 
     In some aspects, administration of a vaccine or composition of the present disclosure to one or more individuals or a human population reduces the incidence of typhoid fever in those individuals or that human population subsequently exposed to pathogenic  S. typhi.    
     In some aspects, administration of a vaccine or composition of the present disclosure to one or more individuals or a human population reduces the incidence of both COVID-19 and typhoid fever in those individuals or that human population subsequently exposed to both pathogenic SARS-CoV-2 and pathogenic  S. typhi.    
     Certain aspects of the present disclosure are directed to an isolated nucleic acid comprising the nucleic acid sequence of SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10, wherein the isolated nucleic acid is codon optimized for expression in  Salmonella typhi  Ty21a. 
     Certain aspects of the present disclosure provide a method of eliciting an immune response in a subject against a SARS-CoV-2 viral antigen or a modification thereof, the method comprising administering one or more doses of a composition or a vaccine disclosed herein to the subject. In some aspects, the composition or vaccine is administered to the mucosa of the subject. In some aspects, the composition or vaccine is administered orally or intranasally to the subject. In some aspects, the composition or vaccine is administered by inhalation. In some aspects, the composition or vaccine induces a humoral antibody response in the subject. In some aspects, the composition or vaccine induces a secretory IgA antibody response in the subject. In some aspects, the composition or vaccine induces a cellular T-cell response in the subject. In some aspects, the subject is a human subject. In some aspects, an immune response specific to a SARS-CoV-2 viral antigen is elicited in the subject by subsequent exposure of the subject to SARS-CoV-2. In some aspects, an immune response to a  Salmonella typhi  antigen is additionally elicited in the subject by subsequent exposure of the subject to pathogenic  Salmonella typhi.    
     Certain aspects of the present disclosure provide a method of eliciting an immune response in a subject against a  Salmonella typhi  antigen, the method comprising administering one or more doses of a composition or a vaccine disclosed herein to the subject. In some aspects, the composition or vaccine is administered to the mucosa of the subject. In some aspects, the composition or vaccine is administered orally or intranasally to the subject. In some aspects, the composition or vaccine is administered by inhalation. In some aspects, the composition or vaccine induces a humoral antibody response in the subject. In some aspects, the composition or vaccine induces a secretory IgA antibody response in the subject. In some aspects, the composition or vaccine induces a cellular T-cell response in the subject. In some aspects, the subject is a human subject. In some aspects, an immune response specific to a  Salmonella typhi  antigen is elicited in the subject by subsequent exposure of the subject to pathogenic  Salmonella typhi . In some aspects, an immune response to a SARS-CoV-2 antigen is additionally elicited in the subject by subsequent exposure of the subject to SARS-CoV-2. 
     Certain aspects of the present disclosure provide a method of treating, preventing, or reducing the incidence of COVID-19 or a SARS-CoV-2 viral infection in one or more subjects, the method comprising administering one or more doses of a composition or a vaccine disclosed herein to each subject. In some aspects, the composition or vaccine is administered to the mucosa of each subject. In some aspects, the composition or vaccine is administered orally or intranasally to each subject. In some aspects, the composition or vaccine is administered by inhalation. In some aspects, the composition or vaccine induces a humoral antibody response in the subject. In some aspects, the composition or vaccine induces a secretory IgA antibody response in the subject. In some aspects, the composition or vaccine induces a cellular T-cell response in the subject. In some aspects, the one or more subjects are human subjects. 
     Certain aspects of the present disclosure provide a method of treating, preventing, or reducing the incidence of typhoid fever in one or more subjects, the method comprising administering one or more doses of a composition or a vaccine disclosed herein to each subject. In some aspects, the composition or vaccine is administered to the mucosa of each subject. In some aspects, the composition or vaccine is administered orally or intranasally to each subject. In some aspects, the composition or vaccine is administered by inhalation. In some aspects, the composition or vaccine induces a humoral antibody response in the subject. In some aspects, the composition or vaccine induces a secretory IgA antibody response in the subject. In some aspects, the composition or vaccine induces a cellular T-cell response in the subject. In some aspects, the one or more subjects are human subjects. 
     Certain aspects of the present disclosure provide a method of treating, preventing, or reducing the incidence of both COVID-19 and typhoid fever in one or more subjects, the method comprising administering one or more doses of a composition or a vaccine disclosed herein to each subject. In some aspects, the composition or vaccine is administered to the mucosa of each subject. In some aspects, the composition or vaccine is administered orally or intranasally to each subject. In some aspects, the composition or vaccine is administered by inhalation. In some aspects, the composition or vaccine induces a humoral antibody response in the subject. In some aspects, the composition or vaccine induces a secretory IgA antibody response in the subject. In some aspects, the composition or vaccine induces a cellular T-cell response in the subject. In some aspects, the one or more subjects are human subjects. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  ( FIG. 1 ) shows the structural components of a coronavirus viral particle (Alsaadi, E. A. J., and J. M. Jones,  Future Virol.  14: 275-86 (2019)). 
         FIG. 2  ( FIG. 2 ) shows the amino acid sequence of a modified SARS CoV-2 Spike glycoprotein ectodomain (eS) (SEQ ID NO: 1) (Wrapp, D., et al.,  Science  367: 1260-3 (2020)). Bolded and underlined=N-glycosylation sites; Solid box=RBD; Bolded and italicized=D614G substitution (Korber B., et al.,  Cell  182(4):812027 (2020)); Hashed box with short lines=GSAS substitution (for RRAR) at the furin cleavage site (residues 682-685); Hashed box with long lines=Proline substitutions at residues 986-987. Additions at the C-terminus: Hashed box with long and short lines=T4 fibritin trimerization motif. 
         FIGS. 3A-3B  ( FIGS. 3A-3B ) show the structure ( FIG. 3A ) and amino acid sequence ( FIG. 3B ) of SARS-CoV2 M protein (SEQ ID NO: 2; GenBank Accession No. MN908947.3; https://COVID-19.uniprot.org/uniprotkb/PODTC5). The region corresponding to amino acids S108 to P123 of the M protein is a conserved region among SARS-CoV-2, SARS-CoV, Bat CoV, and MERS CoV ( FIG. 3B , solid box). 
         FIGS. 4A-4B  ( FIGS. 4A-4B ) show the structure ( FIG. 4A ) and amino acid sequence ( FIG. 4B ) of SARS-CoV-2 N protein (SEQ ID NO: 3; GenBank Accession No. MN908947.3; https://COVID-19.uniprot.org/uniprotkb/PODTC5). Intrinsically disordered regions (IDR) (a.a. 1-44; 182-247; 366-422); N terminal domain (NTD) (a.a. 45-181); Linker region (LKR) (182-247); C-terminal domain (CTD) (248-365). The charged SR rich (striated box) and the nuclear localization signal (NLS, solid box) motifs are shown (FIG. 1 of McBride, R., et al.,  Viruses  6(8), 2991-3018 (2014); Huang, Q., et al., Biochemistry (Mosc) 43: 6059-63 (2004)). 
         FIGS. 5A-5C  ( FIGS. 5A-5C ) show the schematics of the Ty21a-eS ( FIG. 5B ) and Ty21a-eS-AR ( FIG. 5C ) constructs. Chromosomal integration of eS gene is achieved with the Ty21a-eS and Ty21a-eS-AR constructs ( FIGS. 5A-5C ). Secto=gene cassette representing the eS protein. Lpp=constitutive lpp promoter. P ara =arabinose-inducible promoter. 
         FIGS. 6A-6C  ( FIGS. 6A-6C ) show the schematics of the Ty21a-eS-M-N ( FIG. 6B ) and Ty21-eS-M-N-AR ( FIG. 6C ) constructs. Chromosomal integration of the SARS-CoV-2 spike protein ectodomain (eS), M, and N genes is achieved with the Ty21a-eS-M-N and Ty21a-eS-M-N-AR constructs ( FIGS. 6A-6C ). 
         FIGS. 7A-7B  ( FIGS. 7A-7B ) show the plasmid expression of a recombinant SARS-CoV-2 eS in cell pellets (OD 0.15;  FIG. 7A ) and cell supernatant (OD 0.5;  FIG. 7B ). M=Marker, Ty21a, 2.5 ng Spike, 1-4=4 clones. Probed with rabbit anti-SARS-CoV-2/2019-nCo Spike. 
         FIGS. 8A-8B  ( FIGS. 8A-8B ) show that a chromosomally integrated SARS-CoV-2 eS gene fragment expressed eS protein in both cell pellets ( FIG. 8A ) and cell supernatant ( FIG. 8B ) of the Ty21a-S-ecto clone. R=Ty21a with plasmid expressed Spike-ecto. X=chromosomal integrated Ty21a-S-ecto. 
         FIG. 9  ( FIG. 9 ) shows the amino acid sequence of a modified SARS CoV-2 eS (SEQ ID NO: 5). Bolded=the SARS CoV-2 eS. Hashed boxes with long lines=spike modifications. Additions at the C-terminus: Hashed box with long and short lines=T4 fibritin trimerization motif; Hashed box with short lines=hemolysin A (HLYA) signal sequence. 
         FIG. 10  ( FIG. 10 ) shows a schematic for the construction of a Ty21a-RBD-AR construct. As shown, the SARS-CoV-2 receptor binding domain (RBD) was fused to the HlyA secretion signal, and the  Shigella sonnei  acid resistance cassette was integrated into the Ty21a genome at the tviE locus. The schematic also shows locations for primers 27, 37, 89, 64, and 107, which were used in polymerase chain reaction (PCR) amplification. 
         FIGS. 11A-11C  ( FIG. 11A-11C ) show confirmation of the integration via PCR.  FIG. 11A  shows PCR amplification using primers 27 and primer 107, which flank the homologous arms on both the 5′ and 3′ ends of the integration site.  FIG. 11B  shows PCR amplification using primer 27 (flanking the 5′ end of the homologous arm) and primer 37 (inside the insertion and downstream of KanR gene).  FIG. 11C  shows confirmation using primer 64 (downstream of the right homology arm) and primer 89 (inside the insertion and on the 5′ of the RBD). 
     
    
    
     DETAILED DESCRIPTION OF THE DISCLOSURE 
     I. Definitions 
     In order that the present disclosure can be more readily understood, certain terms are first defined. As used in this application, except as otherwise expressly provided herein, each of the following terms shall have the meaning set forth below. Additional definitions are set forth throughout the application. 
     It is to be noted that the term “a” or “an” entity refers to one or more of that entity; for example, “a nucleotide sequence,” is understood to represent one or more nucleotide sequences. As such, the terms “a” (or “an”), “one or more,” and “at least one” can be used interchangeably herein. 
     Furthermore, “and/or” where used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other. Thus, the term “and/or” as used in a phrase such as “A and/or B” herein is intended to include “A and B,” “A or B,” “A” (alone), and “B” (alone). Likewise, the term “and/or” as used in a phrase such as “A, B, and/or C” is intended to encompass each of the following aspects: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone). 
     Similarly, the word “or” is intended to include “and” unless the context clearly indicates otherwise. It is further to be understood that all base sizes or amino acid sizes, and all molecular weight or molecular mass values, given for nucleic acids or polypeptides are approximate, and are provided for description. 
     It is understood that wherever aspects are described herein with the language “comprising,” otherwise analogous aspects described in terms of “consisting of” and/or “consisting essentially of” are also provided. 
     The term “about” is used herein to mean approximately, roughly, around, or in the regions of. When the term “about” is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. Thus, “about 10-20” means “about 10 to about 20.” In general, the term “about” can modify a numerical value above and below the stated value by a variance of, e.g., 10 percent, up or down (higher or lower). 
     The term “expression” as used herein refers to a process by which a polynucleotide produces a gene product, for example, an RNA or a polypeptide. 
     As used herein, the term “recombinant” includes the expression from genes made by genetic engineering or otherwise by laboratory manipulation. 
     “Biosynthetic” as used herein, means produced by a process whereby one or more substrates are converted to more complex products within a living organism or cell. 
     As used herein, “Deoxyribonucleic acid” or “DNA,” is a polynucleotide assembled in a particular sequence that encodes a polypeptide. DNA can include a promoter or secretion signal and/or other transcription or translation control elements operably associated with one or more coding regions. An operable association means that a coding region for a gene product, e.g., a polypeptide, is associated with one or more regulatory sequences in such a way as to place expression of the gene product under the influence or control of the regulatory sequence(s). 
     As used herein, “Nucleic acid” refers to any one or more nucleic acid segments, e.g., DNA fragments, present in a polynucleotide. Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions, codon optimized sequences) and complementary sequences as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and/or deoxyinosine residues (Batzer et al.,  Nucleic Acid Res.  19:5081 (1991); Ohtsuka et al., J. Biol. Chem. 260:2605-8 (1985); Rossolini et al.,  Mol. Cell. Probes  8:91-8 (1994)). 
     “Sequence identity” as used herein refers to a relationship between two or more polynucleotide sequences or between two or more polypeptide sequences. When a position in one sequence is occupied by the same nucleic acid base or amino acid residue in the corresponding position of the comparator sequence, the sequences are said to be “identical” at that position. The percentage “sequence identity” is calculated by determining the number of positions at which the identical nucleic acid base or amino acid residue occurs in both sequences to yield the number of “identical” positions. The number of “identical” positions is then divided by the total number of positions in the comparison window and multi-plied by 100 to yield the percentage of “sequence identity.” Percentage of “sequence identity” is determined by comparing two optimally aligned sequences over a comparison window. In order to optimally align sequences for comparison, the portion of a polynucleotide or polypeptide sequence in the comparison window may comprise additions or deletions termed gaps while the reference sequence is kept constant. An optimal alignment is that alignment which, even with gaps, produces the greatest possible number of “identical” positions between the reference and comparator sequences. The terms “sequence identity” and “identical” are used interchangeably herein. Accordingly, sequences sharing a percentage of “sequence identity” are understood to be that same percentage “identical.” In some embodiments, the percentage “sequence identity” between two sequences can be determined using the program “BLAST 2 Sequences” which was available from the National Center for Biotechnology Information, which program incorporates the programs BLASTN (for nucleotide sequence comparison) and BLASTP (for polypeptide sequence comparison), which programs are based on the algorithm of Karlin and Altschul ( Proc. Natl. Acad. Sci. USA  90(12):5873-7 (1993)). 
     As used herein, a “gene” refers to a locus (or region) of DNA, which is made up of nucleotides that can that can be transcribed into RNA that in turn encodes a polypeptide. 
     As used herein, “gene region” refers to a location within chromosomal DNA that encodes one or more polypeptides of interest (e.g., an antigen). 
     “Gene system” as used herein refers to one or more genes that encode one or more polypeptides which when expressed in concert produce a desired effect. 
     “Variant,” as used herein, refers to a polypeptide that differs from the recited polypeptide due to amino acid substitutions, deletions, insertions, and/or modifications. 
     As used herein, “functional variant” means polypeptides that retain at least some of the properties of the corresponding wild-type polypeptide. For example, in some embodiments, the functional variant of an antigen retains at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% antigenicity and/or protective immunity of the corresponding wild-type antigen. 
     As used herein, “transgenic” refers to an organism or cell that comprises a gene, a gene region, and/or a gene system that has been transferred to it by genetic engineering techniques. 
     “Integrated into” as used herein refers to incorporating a heterologous DNA (e.g., a gene, a gene region, and/or a gene system) into a chromosomal DNA. 
     “Heterologous” as used herein means from a different organism, cell type or species. 
     As used herein, “transformation,” “transfection,” and “transduction” refer to methods of transferring nucleic acid (i.e., a recombinant DNA) into a cell. The transferred nucleic acid can be introduced into a cell via an expression vector such as a plasmid, usually comprising components essential for selection, expression of target gene(s), and/or replication in the host cell. 
     “Stably expressed” as used herein refers to expression of a heterologous gene, a gene region and/or a gene system that has been integrated into chromosomal DNA in a fashion that is reproducible through multiple cell passages and/or under a broad range of physiologic conditions. 
     “Acid tolerance” as used herein refers to the ability of cells to remain viable at low pH, and after exposure to low pH such as passage through the stomach. 
     An “antigen” (also referred to as an immunogen) as used herein is a molecule capable of inducing an immune response in a host organism (e.g., a human) that is specific to that molecule. 
     “Immune response” as used herein means a response in a host organism, e.g., a human, to the introduction of an immunogen (e.g., a transgenic Ty21a of the application) generally characterized by, but not limited to, production of antibodies and/or T cells. In some embodiments, an immune response may be a cellular response such as induction or activation of CD4+ T cells or CD8+ T cells specific for an antigen, a humoral response of increased production of pathogen-specific antibodies, or both cellular and humoral responses. 
     “Vaccine” as used herein is a composition comprising an immunogenic agent (e.g., an immunogen or antigen) and a pharmaceutically acceptable diluent or carrier, optionally in combination with excipient, adjuvant and/or additive or protectant. 
     In certain embodiments, when a vaccine is administered to a subject, the immunogen (e.g., a transgenic Ty21a of the application) stimulates an immune response that will, upon subsequent exposure to an infectious agent, protect the subject from illness or mitigate the pathology, symptoms or clinical manifestations caused by that agent. In some embodiments, a therapeutic (treatment) vaccine is given after infection and is intended to reduce or arrest disease progression. In some embodiments, preventive (prophylactic) vaccine is intended to prevent initial infection or reduce the rate or burden of the infection. 
     “Carrier” as used herein refers to a substance suitable for pharmaceutical use. In some aspects, the carrier renders a composition (e.g., vaccine) suitable for pharmaceutical use. In some embodiments, the carrier is selected from the group consisting of water, PBS, saline, or any combination thereof. In another embodiment the carrier is selected from the group consisting of sucrose, ascorbic acid, amino acid mixture, lactose, magnesium stearate, or any combination thereof. 
     “Conferring protective immunity” refers to providing to a subject (i.e., an individual) or a human population (e.g., at least 10 subjects) the ability to generate an immune response to protect against a disease (e.g., COVID-19 or typhoid fever) caused by subsequent exposure to a pathogen (e.g., a bacteria or virus) such that the clinical manifestations, pathology, or symptoms of disease are reduced during subsequent exposure to the pathogen as compared to a non-treated subject, or such that the rate at which infection, or clinical manifestations, pathology, or symptoms of disease appear within a population are reduced, as compared to a non-treated population. 
     “Human population” as used herein refers to a group of humans which can be represented by a defined number of subjects, e.g., at least two subjects. For example, a human population comprises those individuals participating in a clinical trial, or individuals that have received a vaccine and are then challenged to assess protection. 
     “Dose” as used herein refers to a distinct administration event to a subject. 
     “Immunized” as used herein means sufficiently vaccinated to achieve a protective immune response. 
     As used herein, “stable,” with regard to a vaccine or pharmaceutical composition means the retention of potency for a given period of time. In some aspects, stability of a vaccine strain or a pharmaceutical composition comprising the vaccine strain is determined by assessing viability and/or residual water content over a period of time (e.g., at a certain temperature). In some aspects, viability is measured by growing colonies on an agar plate, counting the colony forming units (CFU), and scoring CFU per unit weight. In some aspects, viability is measured by removing a sample of the foam dried vaccine at one or more time points over a period of time, reconstituting the dried material in an equivalent volume of sterile water to the original pre-foam dried state (ml/mg), and quantitating CFU. In some aspects, CFU are quantitated by plating the reconstituted sample on tryptic soy agar and scoring for CFU/mg in the foam dried product after overnight incubation at 37° C. (Sim, B. K. L., et al., NPJ Vaccines 2:17 (2017)). 
     As used herein, the term “subject” includes any human or non-human animal. For example, the methods and compositions described herein can be used to treat, prevent, or reduce the incidence of a SARS-CoV-2 viral infection in a subject or to elicit an immune response in a subject against a SARS-CoV-2 viral antigen or a modification thereof. The term “non-human animal” includes all vertebrates, e.g., mammals and non-mammals, such as non-human primates, sheep, dog, cow, chickens, amphibians, reptiles, etc. 
     “Treat,” “treatment,” or “treating,” as used herein refers to, e.g., the reduction in severity of a disease or condition; the reduction in the duration of a condition course; the amelioration or elimination of one or more symptoms associated with a disease or condition; the provision of beneficial effects to a subject with a disease or condition, without necessarily curing the disease or condition. 
     “Pharmaceutical formulation” or “pharmaceutical composition,” as used herein, refers to a preparation which is in such form as to permit the biological activity of the active ingredient to be effective, and which contains no additional components which are unacceptably toxic to a subject to which the pharmaceutical formulation or composition would be administered. The pharmaceutical formulation or composition can be sterile. In some aspects, the pharmaceutical formulation or composition is suitable for therapeutic use in a human subject. 
     As used herein, the terms “ug” and “uM” are used interchangeably with “μg” and “μM,” respectively. 
     Any compositions or methods provided herein can be combined with one or more of any of the other compositions and methods provided herein. 
     Units, prefixes, and symbols are denoted in their Systeme International de Unites (SI) accepted form. Numeric ranges are inclusive of the numbers defining the range. Unless otherwise indicated, amino acid sequences are written left to right in amino to carboxy orientation. The headings provided herein are not limitations of the various aspects of the disclosure, which can be had by reference to the specification as a whole. Accordingly, the terms defined immediately below are more fully defined by reference to the specification in its entirety. 
     Various aspects described herein are described in further detail in the following subsections. 
     II. Compositions 
     The present disclosure relates to the preparation and use of  Salmonella typhi  ( S. typhi ) Ty21a vectors to express foreign immunogens, e.g., one or more of the protein immunogens of the SARS-CoV-2 virus (e.g., S, eS, M, N), which have been stably integrated into the Ty21a chromosome. Such constructions provide bivalent or multivalent protection against both typhoid fever (the disease caused by  S. typhi  per se) as well as the disease associated with said foreign immunogen (e.g., COVID-19). 
     A challenge with the spike protein of SARS-CoV-2 is that it contains multiple N-glycosylation sites, positioned in the ectodomain. Therefore, when expressing foreign glycoproteins, e.g., in bacteria, they may fail to be secreted and rather form insoluble aggregates in the intracellular matrix. For example, Chuck et al. (Virus Genes 38:1-9 (2009)) had compared the expression and purification of six different fragments of the SARS CoV spike in  E. coli  and  Pichia pastoris  yeast. Chuck et al. found that the spike fragments expressed in  E. coli  formed aggregates and were insoluble whereas expression in the  P. pastoris  resulted in a significantly higher protein yields, purity and solubility. In certain aspects, the present disclosure provides for the expression of SARS-CoV spike proteins using bacterial Ty21a as disclosed with reduced or no aggregation of the expressed spike proteins (e.g., one or more SARS-CoV-2 viral antigens). 
     In some aspects, the  S. typhi  Ty21a vector used to express foreign immunogens is a transgenic  S. typhi  Ty21a comprising a chromosome with one or more heterologous nucleic acid regions (e.g., one or more biosynthetic gene regions), wherein the heterologous nucleic acid regions encode one or more SARS-CoV-2 viral antigens and are integrated into the  S. typhi  Ty21a chromosome, and wherein the transgenic  S. typhi  Ty21a stably expresses the one or more SARS-CoV-2 viral antigens. 
     In some aspects, the one or more heterologous nucleic acid regions of the transgenic  S. typhi  Ty21a vector encode a portion or all of one or more SARS-CoV-2 structural proteins (e.g., the spike (S) protein, the spike protein ectodomain (eS), the membrane (M) protein, and/or the nucleocapsid (N) protein), either naturally occurring or modified. In some aspects, the one or more heterologous nucleic acid regions are recombineered into the chromosome of Ty21a to stably express these immunogens as vaccine candidates. 
     In some aspects, the one or more heterologous nucleic acid regions of the transgenic  S. typhi  Ty21a vector comprise: (a) a nucleic acid sequence encoding a SARS-CoV-2 spike (S) protein or one or more antigenic fragments thereof, (b) a nucleic acid sequence encoding one or more antigens retaining at least 70% amino acid residue homology to a native SARS-CoV-2 S protein antigen, (c) a nucleic acid sequence encoding a SARS-CoV-2 spike protein ectodomain (eS) or one or more antigenic fragments thereof, (d) a nucleic acid encoding one or more antigens retaining at least 70% amino acid residue homology to a native SARS-CoV-2 eS antigen, (e) a nucleic acid sequence encoding a SARS-CoV2 membrane (M) protein or one or more antigenic fragments thereof, (f) a nucleic acid sequence encoding one or more antigens retaining at least 70% amino acid residue homology to a native SARS-CoV-2 M protein antigen, (g) a nucleic acid sequence encoding a SARS-CoV2 nucleocapsid (N) protein or one or more antigenic fragments thereof, (h) a nucleic acid sequence encoding one or more antigens retaining at least 70% amino acid residue homology to a native SARS-CoV-2 N protein antigen, or (i) any combination thereof. 
     Ty21a encoding a modified SARS-CoV-2 eS protein (Ty21a-eS). In some aspects, the one or more heterologous nucleic acid regions of the transgenic  S. typhi  Ty21a vector encode a modified SARS-CoV-2 eS protein. The SARS-CoV-2 S protein is a large trimeric class I fusion protein that exists in a metastable prefusion conformation that undergoes a substantial structural rearrangement to fuse the viral membrane with the host cell membrane (Li, F.,  Annu Rev Viral.  3: 237-61 (2016); Bosch B. J., et al.,  J Virol.  77: 8801-11 (2003)) of its receptor ACE2 on target cells (high concentrations on epithelial cells of the oral cavity) (Xu H., et al.,  Int J Oral Sci.  12: 8 (2020)). The S protein consists of two subunits, S1 at the N-terminus providing the receptor binding function and S2 at the C-terminus providing fusion activity ( FIG. 1 ) (Belouzard, S., et al.,  Viruses  4: 1011-33 (2012)). For mediation of membrane fusion, cleavage of S/S2 must occur. High infectivity and transmissibility of SARS-COV-2 as compared to SARS-COV has been linked to cleavage efficiency (Coutard, B., et al.,  Antiviral Res.  176: 104742 (2020)). In some aspects, the modified eS comprises a substitution of residues GSAS for RRAR at the furin cleavage site (residues 682-685), for improved accessibility and cleavage (Wrapp, D., et al.,  Science  367: 1260-3 (2020)). In some aspects, Foldon, the T4 fibritin trimerization domain that triggers obligatory trimerization (SEQ ID NO: 12) (Meier, S, et al.,  J Mol Biol.  344: 1051-69 (2004)), is included in the modified eS with the intent of proper trimerization as shown in other systems (Lu, Y., et al.,  Proc Natl Acad Sci USA  111: 125-30 (2014)). Thus, in some aspects, the eS encoded by the one or more heterologous nucleic acid regions contains modifications that stabilize the eS in its prefusion structure, thereby increasing eS expression, secretion, and immunogenicity (Wrapp, D., et al.,  Science  367: 1260-3 (2020); Pallesen, J.,  Proc Natl Acad Sci USA  114: E7348-E7357 (2017)) ( FIG. 2 ). In some aspects, the modified eS comprises a D614G substitution (Korber B., et al.,  Cell  182(4):812027 (2020); Grubaugh, N. D., et al.,  Cell  182(4):794-5 (2020)). There are data on ‘HexaPro’ which show 4 substitutions of prolines (F817P, A892P, A899P, A942P) result in greater expression and stability (Hsieh C-L, et al., bioRxiv: 2020.2005.2030.125484, 2020)). In some aspects, the modified eS comprises F817P, A892P, A899P, and A942P substitutions. In some aspects, the modified eS comprises D614G, F817P, A892P, A899P, and A942P substitutions. In some aspects, the modified eS comprises proline substitutions at residues 986-987. In some aspects, the modified eS comprises a HLYA (SEQ ID NO: 13). In some aspects, the modified eS comprises the amino acids shown in  FIG. 2  (SEQ ID NO: 1) or  FIG. 9  (SEQ ID NO: 5). Ty21a-eS (SEQ ID NO: 6), which expresses a modified eS protein, is constructed and shown in  FIG. 5B . Ty21a-eS-AR (SEQ ID NO: 8), which expresses a modified eS protein and comprises a heterologous AR gene cassette, is constructed and shown in  FIG. 5C . The sequences described herein are disclosed in Table 7. 
     Ty21a encoding a modified SARS-CoV-2 Receptor Binding Domain (RBD) (Ty21a-RBD). In some aspects, the one or more heterologous nucleic acid regions of the transgenic  S. typhi  Ty21a vector encode a modified SARS-CoV-2 RBD. The RBD is part of the ectodomain of the Spike (S) protein. In some aspects, the modified RBD comprises a D614G substitution. In some aspects, the modified RBD comprises a substitution of residues GSAS for RRAR at the furin cleavage site (residues 682-685). In some aspects, the modified RBD comprises proline substitutions at residues 986-987. In some aspects, the modified RBD comprises T4 fibritin trimerization motif at the C-terminus (SEQ ID NO: 12). In some aspects, the modified RBD comprises a HLYA (SEQ ID NO: 13). In some aspects, the modified SARS-CoV-2 RBD comprises the amino acid sequence of SEQ ID NO: 11 (solid box of  FIG. 2 ). 
     Ty21a encoding a SARS-CoV-2 Membrane (M) protein (Ty21a-M). In some aspects, the one or more heterologous nucleic acid regions of the transgenic  S. typhi  Ty21a vector encode the entire SARS-CoV-2 M protein. The M protein is a type III transmembrane protein and is the most abundant protein in the CoV particle and functions in virus assembly at the budding site. It is composed of three parts: a short N-terminal domain situated outside the virion membrane, three transmembrane domains, and a carboxy-terminal domain inside the particle (Hogue, B. G., and Machamer, C. E., 2008 . Coronavirus structural proteins and virus assembly . In Nidoviruses, ed. Perlman, S., Gallagher, T, and Snijder, E., pp. 179-200. Washington, D.C.: ASM Press; Ujike, M., and F. Taguchi,  Viruses  7:1700-25 (2015)). An amphipathic region at the end of the 3 rd  transmembrane domain is conserved in almost all Coronaviridae (Arndt, A. L., et al.,  J Virol.  84: 11418-28 (2010)). In some aspects, the SARS-CoV-2 M protein comprises the amino acids shown in  FIG. 3B  (SEQ ID NO: 2). The SARS-CoV2 M protein is 222 amino acids in length. Ty21a-M, which expresses the entire M protein and includes all M protein epitopes and domains, is shown in  FIGS. 3A-3B  (Grifoni, A., et al.,  Cell  181:1489-1501 (2020); Arndt, A. L., et al.,  J Virol.  84: 11418-28 (2010); Grifoni, A., et al.,  Cell Host Microbe  27: 671-80 (2020)). 
     Ty21 encoding a SARS-CoV-2 Neucleocapsid (N) protein (Ty21a-N). In some aspects, the one or more heterologous nucleic acid regions of the transgenic  S. typhi  Ty21a vector encode the entire SARS-CoV-2 N protein. The N protein, a major structural protein in the virion, chaperones and protects the viral RNA genome (Sturman, L. S., et al.,  J Virol.  33:449-62 (1980)). In some aspects, the SARS-CoV-2 N protein comprises the amino acids shown in  FIG. 4B  (SEQ ID NO: 3). Ty21a-N, which expresses the entire N protein and includes all N protein epitopes and domains, is constructed and shown in  FIGS. 4A-4B  (Grifoni, A., et al.,  Cell Host Microbe  27: 671-80 (2020)). 
     Ty21 encoding a SARS-CoV-2 M protein and a SARS-CoV-2 N (Ty21a-M-N). In some aspects, the one or more heterologous nucleic acid regions of the transgenic  S. typhi  Ty21a vector encode the entire SARS-CoV-2 M protein and the entire SARS-CoV-2 N protein. In some aspects, the SARS-CoV-2 M protein comprises the amino acids shown in  FIG. 3B  (SEQ ID NO: 2, and the SARS-CoV-2 N protein comprises the amino acids shown in  FIG. 4B  (SEQ ID NO: 3). Ty21a-M-N(SEQ ID NO: 7), which expresses the entire M and N proteins and includes all epitopes and domains of the M and N proteins, is constructed and shown in  FIGS. 4A-4B  (Grifoni, A., et al.,  Cell Host Microbe  27: 671-80 (2020)). 
     Ty21 encoding a modified SARS-CoV-2 eS protein, a SARS-CoV-2 M protein, and a SARS-CoV-2 N protein (Ty21a-eS-M-N). In some aspects, the one or more heterologous nucleic acid regions of the transgenic  S. typhi  Ty21a vector encode a modified SARS-CoV-2 eS protein, a full-length SARS-CoV-2 M protein, and a full-length SARS-CoV-2 N protein. Ty21a-eS-M-N, which expresses a modified eS protein, the entire M protein, and the entire N protein, is constructed and shown in  FIG. 6B . 
     In some aspects, the transgenic  S. typhi  Ty21a vectors disclosed herein further comprise a heterologous acid resistance (AR) gene cassette (e.g., a heterologous AR biosynthetic gene system) integrated into the  S. typhi  Ty21a chromosome. Ty21a-eS-AR (SEQ ID NO: 8), which expresses a modified eS protein, the entire M protein, and the entire N protein and comprises a heterologous AR gene cassette, is constructed and shown in  FIG. 5C . Ty21a-M-N-AR (SEQ ID NO: 9), which expresses the entire M protein, and the entire N protein and comprises a heterologous AR gene cassette, is constructed. Ty21a-es-M-N-AR (SEQ ID NO: 10), which expresses a modified eS protein, the entire M protein, and the entire N protein and comprises a heterologous AR gene cassette, is constructed and shown in  FIG. 6C . 
     Integration of a heterologous AR gene cassette provides acid stability and enhances viability of recombinant Ty21a as it passes through the stomach where conditions are acidic, thereby providing for more stable gene expression. This can also eliminate the need for gelatin capsules or liquid formulations and provides temperature stabilization and extended shelf life. Non-limiting examples of heterologous AR biosynthetic gene system are disclosed in U.S. Pat. No. 10,695,415, which is incorporated by reference herein in its entirety, and include  Shigella  glutamine-glutamate AR gene systems. In some aspects, the immunogenicity of the  S. typhi  Ty21a vectors disclosed herein may be enhanced by improving its ability to withstand low pH passage through the stomach by incorporating a  Shigella  glutamine-glutamate AR gene system. 
     In some aspects, the heterologous AR gene cassette comprises a gene selected from the group consisting of: a YbaS gene (SEQ ID NO: 15) or a fragment thereof, a GadB gene (SEQ ID NO: 17) or fragment thereof, a GadC gene (SEQ ID NO: 19) or fragment thereof, a GadA gene (SEQ ID NO: 22) or fragment thereof, and any combination thereof. In some aspects, the heterologous AR gene cassette comprises a sequence having at least 80%, 85%, 90%, 95%, or 100% sequence identity to one or more nucleic acid sequences selected from: SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 20, and SEQ ID NO: 22. In some aspects, the transgenic  S. typhi  Ty21a vectors comprising the heterologous AR gene cassette are more acid stable at pH 2.5 than  Salmonella typhi  Ty21a without the integrated AR gene cassette. 
     In some aspects, the  S. typhi  Ty21a vectors of the present disclosure are formulated as a composition or vaccine. In some aspects, the composition or vaccine comprises a carrier that renders the composition or vaccine suitable for pharmaceutical use. In some aspects, the composition or vaccine is suitable for oral or inhaled administration (e.g., nasal aerosol administration). In some aspects, the  S. typhi  Ty21a vectors of the present disclosure are formulated as a composition or vaccine for oral or inhaled administration (e.g., nasal aerosol administration) and comprise an integrated heterologous AR cassette. In some aspects, the  S. typhi  Ty21a vectors of the present disclosure are formulated as a composition or vaccine for oral or inhaled administration (e.g., nasal aerosol administration) without said integration of a heterologous AR cassette. 
     In some aspects, the vaccine is protective against COVID-19. In some aspects, the vaccine is protective against typhoid fever. In some aspects, the vaccine is protective against COVID-19 and typhoid fever. 
     In some aspects, administration of the vaccine to one or more individuals or a human population reduces the incidence of COVID-19 in the one or more individuals or the human population when subsequently exposed to pathogenic SARS-CoV-2. In some aspects, administration of the vaccine to one or more individuals or a human population reduces the incidence of typhoid fever in the one or more individuals or the human population when subsequently exposed to pathogenic  S. typhi . In some aspects, administration of the vaccine to one or more individuals or a human population reduces the incidence of both pathogenic SARS-CoV-2 and typhoid fever in the one or more individuals or the human population when subsequently exposed to pathogenic SARS-CoV-2 and pathogenic  S. typhi.    
     In some aspects, the vaccine is foam dried for stabilization at room temperature. In some aspects, the vaccine is at least 90% stable, at least 95% stable, at least 99% stable, or 100% stable at room temperature for at least one month. In some aspects, the vaccine is at least 40% stable, at least 50% stable, at least 60% stable, at least 70% stable, at least 80% stable, at least 90% stable, at least 95% stable, at least 99% stable, or 100% stable at room temperature for at least one year. In some aspects, the vaccine is at least 40% stable, at least 50% stable, at least 60% stable, at least 70% stable, at least 80% stable, at least 90% stable, at least 95% stable, at least 99% stable, or 100% stable at room temperature for at least two years. In some aspects, the vaccine has a shelf-life of 6-24 months, 1-2 years, 1-5 years, 1-10 years, or 5-10 years at 4° C. In some aspects, stability of a foam dried vaccine is determined by assessing viability and/or residual water content over a period of time (e.g., for at least one year at room temperature or for 6-24 months, 1-2 years, 1-5 years, 1-10 years, or 5-10 years at 4° C.). In some aspects, viability is measured by growing colonies on an agar plate, counting the colony forming units (CFU), and scoring CFU per unit weight. In some aspects, viability is measured by removing a sample of the foam dried vaccine at one or more time points over a period of time, reconstituting the dried material in an equivalent volume of sterile water to the original pre-foam dried state (ml/mg), and quantitating CFU. In some aspects, CFU are quantitated by plating the reconstituted sample on tryptic soy agar and scoring for CFU/mg in the foam dried product after overnight incubation at 37° C. (Sim, B. K. L., et al.,  NPJ Vaccines  2:17 (2017)). 
     The disclosure further provides an isolated nucleic acid comprising the nucleic acid sequence of SEQ ID NO: 6 (Ty21a-eS construct), SEQ ID NO: 7 (Ty21a-M-N construct), SEQ ID NO: 8 (Ty21a-eS-AR construct), SEQ ID NO: 9 (Ty21a-M-N-AR construct), or SEQ ID NO: 10 (Ty21a-eS-M-N-AR construct), wherein the isolated nucleic acid is codon optimized for expression in  Salmonella typhi  Ty21. 
     III. Methods of Use 
     The disclosure provides methods of eliciting an immune response in a subject against a SARS-CoV-2 viral antigen or a modification thereof comprising administering one or more doses of a composition or vaccine disclosed herein to the subject. In some aspects, an immune response specific to a SARS-CoV-2 antigen is elicited in the subject by subsequent exposure of the subject to SARS-CoV-2. In some aspects, an immune response to a  Salmonella typhi  antigen is additionally elicited in the subject by subsequent exposure of the subject to pathogenic  Salmonella typhi.    
     The disclosure further provides methods of eliciting an immune response in a subject against a  Salmonella typhi  antigen comprising administering one or more doses of a composition or a vaccine disclosed herein to the subject. In some aspects, an immune response specific to a  Salmonella typhi  antigen is elicited in the subject by subsequent exposure of the subject to pathogenic  Salmonella typhi . In some aspects, an immune response to a SARS-CoV-2 antigen is additionally elicited in the subject by subsequent exposure of the subject to SARS-CoV-2. 
     The disclosure further provides methods of eliciting an immune response in a subject against both a SARS-CoV-2 viral antigen or a modification thereof and a  Salmonella typhi  antigen comprising administering one or more doses of a composition or vaccine disclosed herein to the subject. In some aspects, an immune response to a SARS-CoV-2 antigen is elicited in the subject by subsequent exposure of the subject to SARS-CoV-2, and wherein an immune response specific to a  Salmonella typhi  antigen is elicited in the subject by subsequent exposure of the subject to pathogenic  Salmonella typhi.    
     The disclosure further provides methods of treating, preventing, or reducing the incidence of COVID-19 or a SARS-CoV-2 viral infection in one or more subjects comprising administering one or more doses of a composition or a vaccine disclosed herein to each subject. 
     The disclosure further provides methods of treating, preventing, or reducing the incidence of typhoid fever in one or more subjects comprising administering one or more doses of a composition or vaccine disclosed herein to each subject. 
     The disclosure further provides methods of treating, preventing, or reducing the incidence of both COVID-19 and typhoid fever in one or more subjects comprising administering one or more doses of a composition or vaccine disclosed herein to each subject. 
     In some aspects, the composition or vaccine induces a humoral antibody response in the subject. In some aspects, the composition or vaccine induces a secretory IgA antibody response in the subject. In some aspects, the composition or vaccine induces a cellular T-cell response in the subject. 
     In some aspects, the administration route is oral or nasal. In some aspects, the administration (e.g., immunization) and/or infection route is oral (per os). In some aspects, the administration (e.g., immunization) route is nasal. In some aspects, the composition or vaccine is administered by inhalation. The major barrier for a live oral vaccine is the extreme low pH the vaccine encounters in the stomach.  Salmonella  does not survive well under conditions &lt;pH 3, while most  E. coli  strains and  Shigella  spp. can maintain viability in stomach for several hours (Lin, J., et al.,  J Bacteriol.  177(14):4097-104 (1995); Gorden, J. and P. L. Small,  Infect Immun,  61(1):364-7 (1993)). The ability of  Shigella , and the inability of  Salmonella , to survive at low pH may partially explain why only 10-100  Shigella  cells are sufficient to cause infection, while the infective dose for  Salmonella  spp. ranges at ˜10 5  CFU (U.S. Pat. No. 10,695,415).  Salmonella  expresses acid tolerance response (ATR) genes in response to moderately low pH, which protect the cells from acid challenge as low as pH 3 (Lin, J., et al. (1995); Audia, J. P., et al.,  Int J Med Microbiol.  291(2):97-106 (2001); Foster, J. W.,  J Bacteriol.  173(21):6896-902 (1991); Foster, J. W.,  J Bacteriol,  175(7):1981-7 (1993); Foster, J. W.,  Crit Rev Microbiol,  21(4):215-37 (1995)). Ty21a inherited an rpoS mutation from its parental strain Ty2 (Robbe-Saule, V. and F. Norel,  FEMS Microbiol Lett.  170(1):141-3 (1999)), and carries other less well defined mutations from the random mutagenesis process during strain attenuation (Germanier, R. and E. Fuer,  J Infect Dis.  131(5):553-8 (1975)). Perhaps because of these mutations, Ty21a develops a poor ATR response and is particularly sensitive to low pH (Hone, D. M., et al.,  Vaccine  12(10):895-8 (1994)). However, Ty21a viability is important for vaccine efficacy, as a previous report demonstrated that, when administered orally, live Ty21a elicited stronger and longer lasting immune responses in humans than killed Ty21a (Kantele, A., et al.,  Microb Pathog.  10(2):117-26 (1991)). To facilitate the journey from mouth to ileum without being eliminated in the gastric acid environment, Ty21a is presently placed in enteric-coated capsules that withstand gastric low pH. Additionally, when administered as a liquid with a buffer, Ty21a was more protective (Levine, M. M., et al.,  Vaccine  17 Suppl 2:S22-7 (1999)). On the other hand, capsules are child-unfriendly and adversely impact compliance. Also, the Ty21a liquid formulation has been commercially unsuccessful, in part because it is cumbersome. 
     To obviate the need for special capsules or liquid formulation in buffer, increase bioavailability, reduce dosage requirements, and increase immunogenicity, Ty21a has been rendered acid tolerant (also referred to as acid resistant). There are 5 bacterial acid resistance (AR) pathways, which utilize excess protons to decarboxylate a specific amino acid (e.g. aspartic acid, phenylalanine, lysine, or glutamic acid), and an antiporter that transports the decarboxylated product extracellularly (Waterman, S. R. and P. L. Small,  FEMS Microbiology Letters  225(1):155-60 (2003); Bhagwat, A. A. and M. Bhagwat,  FEMS Microbiology Letters  234(1):139-47 (2004)). The ability of  E. coli, Shigella, Listeria monocytogenes  and  Lactococcus lactis  to withstand extreme acidic pH (below pH 2.5) primarily relies on the most potent AR system, AR2, also known as the glutamate-dependent acid resistance (GDAR) pathway (Lin, J., et al. (1995); Lin, J., et al., Appl Environ Microbiol. 62(9):3094-100 (1996)). AR2 consists of the enzyme glutamate decarboxylase (GAD), encoded by the homologous genes gadA and gadB, and a membrane bound antiporter, encoded by the gene gadC. The two GAD isoforms, GadA and GadB, consume an intracellular proton to decarboxylate glutamate, producing γ-amino butyric acid (GABA) and CO 2  (Audia, J. P., et al. (2001); Brenneman, K. E., et al.,  J Bacteriol.  195(13):3062-72 (2013); De Biase, D. and E. Pennacchietti,  Mol Microbiol.  86(4):770-86 (2012); Merrell, D. S. and A. Camilli,  Curr Opin Microbiol.  5(1):51-5 (2002); Zhao, B. and W. A. Houry,  Biochem Cell Biol.  88(2):301-14 (2010)), while GadC pumps the substrate (glutamate) and product (GABA) in and out of the cell (U.S. Pat. No. 10,695,415; De Biase, D. and E. Pennacchietti,  Mol Microbiol,  86(4):770-86 (2012)). Genes of the two GAD isoforms are located in two distinct chromosomal loci, with gadB and gadC transcribed as a dicistronic operon, and gadA from a separate gene locus (De Biase, D. and E. Pennacchietti (2012); Hersh, B. M., et al.,  J Bacteriol.  178(13):3978-81 (1996); Smith, D. K., et al.,  J Bacteriol.  174(18):5820-6 (1992)). GadA and GadB are highly similar, with sequence identity of 96.5% at nucleotide level and 98.7% at protein level. Deletion of either gadA or gadB does not affect the cell&#39;s AR ability, suggesting these two isozymes are functionally redundant. Newly discovered glutamine-dependent AR system in  E. coli  (Lu, P., et al.,  Cell Res.  23(5):635-44 (2013)) and  Lactobacillus reuteri  (Datsenko, K. A. and B. L. Wanner,  Proc Natl Acad Sci USA  97(12):6640-5 (2000)) have been reported. In  E. coli , a previously uncharacterized bacterial glutaminase A, encoded by the gene ybaS, converts glutamine into glutamate in acidic conditions and releases an ammonium, neutralizing an intracellular proton (Lu, P., et al. (2013)). Interestingly, the antiporter responsible for substrate-product transportation across cell membrane is also GadC (Lu, P., et al. (2013)), consistent with the broad substrate specificity of GadC in vitro (Ma, D., et al.,  Nature  483(7391):632-6 (2012)). Because the product of glutaminase is precisely the substrate of GAD, the YbaS-GadC and GAD-GadC systems work in concert to convert a glutamine molecule into GABA, neutralizing two protons in the cell (U.S. Pat. No. 10,695,415; Lu, P., et al. (2013)), thereby doubling the proton-reducing capacity of the system. This concerted AR system functions more efficiently than the GAD-GadC system alone. 
     In addition to providing multivalent protection against both typhoid fever and SARS-CoV-2 antigens, in some embodiments, integration of an acid resistance cassette eliminates the need for gelatin capsules or liquid formulations and provides temperature stabilization and extended shelf life. 
     The practice of the present disclosure will employ, unless otherwise indicated, conventional techniques of cell biology, cell culture, molecular biology, transgenic biology, microbiology, recombinant DNA, and immunology, which are within the skill of the art. Such techniques are explained fully in the literature. See, for example, Sambrook et al., ed. (1989) Molecular Cloning A Laboratory Manual (2nd ed.; Cold Spring Harbor Laboratory Press); Sambrook et al., ed. (1992) Molecular Cloning: A Laboratory Manual, (Cold Springs Harbor Laboratory, NY); D. N. Glover ed., (1985) DNA Cloning, Volumes I and II; Gait, ed. (1984) Oligonucleotide Synthesis; Mullis et al. U.S. Pat. No. 4,683,195; Hames and Higgins, eds. (1984) Nucleic Acid Hybridization; Hames and Higgins, eds. (1984) Transcription And Translation; Freshney (1987) Culture Of Animal Cells (Alan R. Liss, Inc.); Immobilized Cells And Enzymes (IRL Press) (1986); Perbal (1984) A Practical Guide To Molecular Cloning; the treatise, Methods In Enzymology (Academic Press, Inc., N.Y.); Miller and Calos eds. (1987) Gene Transfer Vectors For Mammalian Cells, (Cold Spring Harbor Laboratory); Wu et al., eds., Methods In Enzymology, Vols. 154 and 155; Mayer and Walker, eds. (1987) Immunochemical Methods In Cell And Molecular Biology (Academic Press, London); Weir and Blackwell, eds., (1986) Handbook Of Experimental Immunology, Volumes I-IV; Manipulating the Mouse Embryo, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., (1986); Crooks, Antisense drug Technology: Principles, strategies and applications, 2nd Ed. CRC Press (2007) and in Ausubel et al. (1989) Current Protocols in Molecular Biology (John Wiley and Sons, Baltimore, Md.). 
     All of the references cited above, as well as all references cited herein and the amino acid or nucleotide sequences (e.g., GenBank numbers and/or Uniprot numbers), are incorporated herein by reference in their entireties. 
     The following examples are offered by way of illustration and not by way of limitation. 
     IV. Examples 
     IV.A. Example 1: Development of Transgenic Ty21a Strains 
     The eS sequence ( FIG. 2 ) was synthesized with intended modifications in plasmid puc57 separately (Genewiz, South Plainfield, N.J.) for recombineering into the Ty21 chromosome at the tviE locus (Ty21a-eS;  FIGS. 5A-5B ). The k-red recombineering technology (Datsenko, K. A., and B. L. Wanner,  Proc Natl Acad Sci USA  97:6640-5 (2000); Dharmasena, M. N., et al.,  Int J Med Microbiol.  303:105-13 (2013)) was used along with methods previously described (Wu, Y., et al.,  J Infect Dis.  215: 259-68 (2017); Sim, B. K. L., et al.,  Vaccine  38: 5123-30 (2020). The constitutive lpp promoter (Lpp-outer membrane lipoprotein) (Inouye, S. and M. Inouye,  Nucleic Acids Res.  13:3101-10 (1985)) was used to provide high expression, and the hemolysin A type 1 secretion system (Hly) was used (Thomas S., et al.,  Biochimica et Biophysica Acta  ( BBA )— Molecular Cell Research  1843:1629-41 (2014)) as the secretion signal ( FIG. 5B ). Creation of the acid stable counterpart candidates (Ty21a-eS-AR;  FIG. 5C ) included the concerted glutaminase-GAD AR systems under the control of AraC the transcriptional regulator for ara operon and the arabinose-inducible promoter (P ara ) (Wu, Y., et al.,  J Infect Dis.  215: 259-68 (2017); Dorman, C. J.,  EcoSal Plus  1(1), DOI: 10.1128/ecosalplus.8.9.3 (2004)) ( FIG. 5C ). A seed bank (20 glycerol stabilate vials stored at −80° C.) was generated for each of the transgenic Ty21a-eS and Ty21a-eS-AR strains. 
     Ty21a-RBD, Ty21a-RBD-AR, Ty21a-M, Ty21a-M-AR, Ty21a-N, Ty21a-N-AR, Ty21a-M-N, Ty21a-M-N-AR, Ty21a-eS-M-N ( FIG. 6B ), and Ty21a-eS-M-N-AR ( FIG. 6C ) constructs were synthesized for recombineering into the Ty21 chromosome at the tviE locus (Ty21a-eS;  FIGS. 5A and 6A ) using an approach similar to the approach described above for the Ty21a-eS and Ty21a-eS-AR constructs. A seed bank (20 glycerol stabilate vials stored at −80° C.) was generated for each of the Ty21a-RBD, Ty21a-RBD-AR, Ty21a-M, Ty21a-M-AR, Ty21a-N, Ty21a-N-AR, Ty21a-M-N, Ty21a-M-N-AR, Ty21a-eS-M-N, and Ty21a-eS-M-N-AR transgenic strains. 
     IV.B. Example 2: Characterization of Transgenic Ty21a Strains 
     i) Secreted expression of recombinant eS from transgenic Ty21a-eS and Ty21a-eS-AR strains was verified by immunoblot using antibodies against S protein and RBD (ProSci, Poway, Calif.); ii) Binding capability of recombinant eS was assessed in ELISAs with binding to recombinant ACE2 as capture antigen (Acro Biosystems, Newark, Del.); iii) Strains were characterized by total genome analysis; iv) Expression stability of chromosomally integrated eS gene fragment was confirmed by characterizing after culturing through 200 generations of growth; v) Strains were microbiologically characterized by analytical profile index (API) analysis-20E Enteric ID system to discriminate for family Enterobacteriaceae; vi) Vaccine strains for known Ty21a phenotypes (galactose-induced cell lysis, isoleucine-valine deficiency, cysteine-deficiency, tryptophan-deficiency, weak H 2 S production) were assessed. 
     Transgenic Ty21a-RBD, Ty21a-RBD-AR, Ty21a-M, Ty21a-M-AR, Ty21a-N, Ty21a-N-AR, Ty21a-M-N, Ty21a-M-N-AR, Ty21a-eS-M-N, and Ty21a-eS-M-N-AR strains were also characterized for i) secreted expression of recombinant eS or RBD, ii) binding capability of recombinant eS, iii) by total genome analysis, iv) expression stability of chromosomally integrated eS, M, and/or N gene fragments, v) API to discriminate for family Enterobacteriaceae and/or vi) known Ty21a phenotypes (galactose-induced cell lysis, isoleucine-valine deficiency, cysteine-deficiency, tryptophan-deficiency, weak H 2 S production). 
     IV.C. Example 3: Assessment of Proper Folding of eS or RBD 
     The RayBio COVID-19 Spike-ACE2 binding assay II (RayBio, Peachtree Corners, Ga.) was used to characterize the binding capability of secreted recombinant eS or RBD to the Angiotensin I Converting Enzyme 2 (ACE2) receptor complex. The assay used a 96-well plate coated with recombinantly-expressed human ACE2 protein. The supernatant of Ty21a-eS, Ty21a-eS-AR, Ty21a-RBD, Ty21a-RBD-AR, Ty21a-eS-M-N, or Ty21a-eS-M-N-AR was added to the wells in the presence of recombinant Fc tagged SARS-CoV-2 spike RBD protein. Unbound RBD was removed with washing. HRP-conjugated anti-mouse IgG was then applied to the wells in the presence of 3,3′,5,5′-tetramethylbenzidine (TMB) substrate. The HRP-conjugated IgG bound to Fc tagged RBD protein and reacted with the TMB solution, producing a blue color that was proportional to the amount of bound RBD. 
     IV.D. Example 4: Foam Drying of Vaccine 
     Foam drying vaccine. Recombineered Ty21a-eS-AR strain was grown in CY medium supplemented with 0.02% galactose to early stationary phase (Sim, B. K. L., et al.,  NPJ Vaccines,  2:17 (2017); Ohtake, S., et al.,  Vaccine  29(15):2761-71 (2011)). The organisms were harvested, resuspended in CY medium, and mixed 1:1 with a 2× foam-drying protective agent (FPA) cocktail in CY medium to achieve a final concentration of 25% trehalose (w/w), 5% fish gelatin (w/w), 0.5% methionine (w/w), and 0.34% potassium phosphate, pH 8.0. Five mL aliquots of the recombineered Ty21a formulation were dispensed into 100-mL Schott vials (Allergy Laboratories, Oklahoma City, Okla.). Foam drying was performed using a Virtis Advantage XL-70 lyophilizer with the following program: Decreasing pressure from 760 Torr to 80 Torr over 15 min at 10° C., holding at 80 Torr for 45 min, decreasing pressure to 5 Torr over 20 min, and a further decrease to 1 Torr at 22° C. with holding for 2 h and further reduction in pressure to &lt;100 m Torr and holding at 15° C. for 48 h. Stoppering was performed in the lyophilizer after argon purging. Residual water content was determined using the Karl Fischer method on a Mettler-Toledo Stromboli balance. 
     IV.E. Example 5: Assessment of Stability Profile of Foam Dried Vaccines 
     Establishing the stability profile of Ty21a-eS-AR using a foam drying process. Stability of the foam dried products (e.g., stability at room temperature (20-25° C.)) is being assessed by residual moisture content and viability (most recent time point at 4 months) (Sim, B. K. L., et al.,  NPJ Vaccines,  2:17 (2017)). Viability was measured by removing a sample of the foam dried product at one or more time points over a period of time, reconstituting the dried material in an equivalent volume of sterile water to the original pre-foam dried state (ml/mg), and quantitating CFU. CFU were quantitated by plating the reconstituted sample on tryptic soy agar and scoring for CFU/mg in the foam dried product after overnight incubation at 37° C. (Sim, B. K. L., et al., NPJ Vaccines 2:17 (2017)). 
     IV.F. Example 6: Immunogenicity of Foam Dried Ty21a-eS and Ty21a-eS-AR Vaccines 
     Immunogenicity of foam dried vaccine candidates. Mice, hamsters, and/or NHPs (rhesus macaques) are immunized and assessed for: 1) antibody levels, 2) neutralization activity of antibodies, 3) systems serology and functional profiling (NHPs only), 4) antibody dependent enhancement, 5) cellular immune responses (mice and NHPs), 6) VE (hamsters and NHPs). 
     Immunogenicity in mice (Table 1).  S. typhi  (Ty21a) can only infect humans (Spanò, S.,  Adv Exp Med Biol.  915:283-94 (2016)). Thus, the true evaluation of the immunogenicity of Ty21a-eS-AR by the oral route can only be assessed in humans. Nevertheless, with these considerations, is important to assess the humoral and cellular responses induced by immunization to demonstrate that the protein produced by Ty21a-es-AR induces neutralizing antibodies and relevant cellular immune responses. BALB/c mice are immunized by intraperitoneal (IP) injection on days 0, 14 and 28 with 5×10 7  CFU of Ty21a-eS-AR. The control is Ty21a. (N=10 mice/group). This dose of recombinant Ty21a stimulates robust immune responses (Wu, Y., et al.,  J Infect Dis.  215:259-68 (2017); Sim, B. K. L., et al.,  NPJ Vaccines  2:17 (2017)). 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Mice receive 3 doses at 2-week intervals. Serum and 
               
               
                 PBMCs collected pre-immunization and 10 days post 
               
               
                 dose 3. Splenocytes collected 10 days post dose 3. 
               
            
           
           
               
               
               
               
               
               
            
               
                 Group 
                 N (M/F) 
                 Immunogen 
                 Volume 
                 # doses 
                 Dose (CFU) 
               
               
                   
               
               
                 1 
                 10 (5/5) 
                 Ty21a-eS-AR 
                 0.2 mL 
                 3 
                 5 × 10 7  CFU 
               
               
                 2 
                 10 (5/5) 
                 Ty21a-eS 
                 0.2 mL 
                 3 
                 5 × 10 7  CFU 
               
               
                 3 
                 10 (5/5) 
                 Ty21a 
                 0.2 mL 
                 3 
                 5 × 10 7  CFU 
               
               
                   
               
            
           
         
       
     
     ELISA. Spike proteins for ELISA assessments are purchased (RayBiotech, GA, Sino Biological, and/or provided by Dr. Galit Alter of Ragon Institute). Subclass IgG assessments determine Th1 (IgG2a, IgG3) and Th2 (IgG1) responses. 96-well plates are coated with 1 μg/ml SARS-CoV-2 Spike (S) protein and serial dilutions of serum diluted in casein block are added with standard blocking and washing steps in between (Smith, T. R. F., et al.,  Nat Commun.  11:2601 (2020)). IgA levels are also measured using recombinant S protein as capture antigen and secondary goat anti-mouse biotin labeled IgA, cat #abx134852 (Abbexa, Houston, Tex.). ELISA antibody levels are defined as the serum dilution at which the optical density (OD) is 0.5 (OD 0.5) (Epstein, J. E., et al.,  Science  334: 475-80 (2011)). 
     Neutralization assay. An authentic SARS CoV-2 plaque reduction neutralization assay is used to quantify neutralization, as described (Zhang, Q., et al.,  Nano Letters  20:5570-4 (2020)). Briefly, a known amount of virus is incubated for 60 minutes with various dilutions of serum from immunized animals. Following incubation, the viable virus is quantified by plaque assay. Dilutions are performed in triplicate and each of the dilutions is assayed in triplicate. 
     Antibody-dependent enhancement (ADE) studies. The ADE assay are done in HL-CZ, promonocyte or Raji cell lines that are susceptible to SARS-CoV as described previously (Wang, S. F., et al.,  Biochem Biophys Res Commun.  451:208-14 (2014)). Cells are infected with pseudotyped viruses in the presence of control or test sera at 10- to 2000-fold dilutions and incubated at 37° C. for 48 h. Supernatants and cell lysates are collected and viral titers determined by quantitative RT-PCR or luminescence analyses to determine if test sera increases the viral load in cells. Cell apoptosis is determined by Annexin V staining as described previously (Wang, S. F., et al.,  Biochem Biophys Res Commun.  451:208-14 (2014)). 
     Cellular responses. Mice are sacrificed 10 days after the 3 rd  dose and splenocytes acquired. ELISpot assays are performed to assess IFN-γ and IL-4 responses. 2×10 5  fresh splenocytes/well are stimulated for 20 h with pools of 15 amino acid peptides overlapping by 9 amino acids and representing the eS (or RBD) as described and provided by Dr. Alex Sette (Smith, T. R. F., et al.,  Nat Commun.  11:2601 (2020)). ELISpot kits (Mabtech) are used per manufacturer&#39;s protocols. 
     IV.G. Example 7: Immunogenicity and Vaccine Efficacy of Ty21a-eS-AR 
     Immunogenicity and vaccine efficacy (VE) in the Syrian hamster model (Chan, J., et al.,  Clin Infect Dis.  71(9):2428-46 (2020)). The dosing regimen follows that in humans for Ty21a (administered orally) (Vivotif [package insert-USA]. Crucell Switzerland LTD; September 2013. http://www.fda.gov/downloads/BiologicsBloodVaccines/Vaccines/ApprovedProducts/U CM142807.pdf). Hamsters and NHPs are immunized by the intranasal (IN) route, instead of orally, because the oral route for  S. typhi /Ty21a is only reliable in humans. Groups of 10 (M=F), 6-10-week-old hamsters are dosed with 5×10 7  CFU IN on days 1, 3, and 5 (Table 2) at BioQual Inc (https://www.bioqual.com). Sera are collected prior to immunizations and 28 days after the 3 rd  immunization (day 33, just before challenge) and antibody assays performed as above. Challenge strain SARS-CoV-2 USA-WA1/2020 stock from BEI Resource (NR-52281; Lot #70033175) is used. Virus inoculum is prepared per Bioqual SOPs by serial dilution in PBS as required to reach the intended dose level of 10 5  pfu in 0.1 mL. Administration of virus is conducted where 0.05 mL of the viral inoculum will be administered dropwise into each nostril, 0.1 mL per animal 4 weeks after the 3 rd  dose (Chan, J., et al.,  Clin Infect Dis.  71(9):2428-46 (2020)). See below for VE assessments. Antibody assays is done as in SA1.5.1, but with reagents appropriate for hamsters (e.g. for ELISA, My Biosource.com, San Diego, Calif., Cat #MBS053809). Cellular immunology assays are not done. 
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 Syrian hamsters are immunized 3 times at 2-day intervals with Ty21a-es-AR or Ty21a 
               
               
                 by IN instillation of 0.05 mL per naris on days 1, 3, and 5, and challenged by 
               
               
                 IN inoculation of 0.05 mL per naris of SARS-CoV-2 28 days later on day 33. 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                 Total/Dose 
                 Total Challenge 
               
               
                 Group 
                 N (M/F) 
                 Immunogen 
                 Volume 
                 # Doses 
                 (CFU) 
                 Dose (PFU) 
               
               
                   
               
               
                 1 
                 10 (5/5) 
                 Ty21a-eS-AR 
                 0.1 mL 
                 3 
                 5 × 10 7   
                 10 5   
               
               
                 2 
                 10 (5/5) 
                 Ty21a (control) 
                 0.1 mL 
                 3 
                 5 × 10 7   
                 10 5   
               
               
                   
               
            
           
         
       
     
     Immunogenicity and VE in non-human primates (NHP) Rhesus macaques (Chandrashekar, A., et al.,  Science  369(6505):812-7 (2020)). Groups of 3, 8-year-old rhesus macaque non-human primates (NHPs) are dosed with 5×10 8  CFU IN on days 1, 3, 5, and 7 (Table 3) (the regimen in humans for Ty21a taken orally) at BioQual Inc. (https://www.bioqual.com/). Sera and peripheral blood mononuclear cells (PBMCs) are collected prior to first immunization and 14 days (PBMCs) and 28 days (sera, just before challenge) after the 4 th  immunization, and antibody and T cells assays are done as described below. Challenge strain SARS-CoV-2 USA-WA1/2020 stock from BEI Resource are diluted in PBS to the desired concentration, and the inoculum are back-titrated to verify the dose given. DMEM containing 10 5  PFU of SARS-CoV-2 in 1 mL are IN inoculated in 0.5 mL per naris as described 4 weeks after the 3 rd  dose. See below for VE assessments. 
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 NHPs are immunized 4 times at 2-day intervals with Ty21a-es-AR or Ty21a 
               
               
                 by IN instillation of 0.5 mL per naris on days 1, 3, 5, and 7 (the 
               
               
                 regimen in humans for Ty21a taken orally) challenged by IN inoculation 
               
               
                 of 0.5 mL per naris of SARS-CoV-2 28 days later on day 35. 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                 Total/Dose 
                 Total Challenge 
               
               
                 Group 
                 N (M/F) 
                 Immunogen 
                 Volume 
                 # Doses 
                 (CFU) 
                 Dose (PFU) 
               
               
                   
               
               
                 1 
                 4 (2/2) 
                 Ty21a-eS-AR 
                 1 mL 
                 4 
                 5 × 10 8   
                 10 5   
               
               
                 2 
                 4 (2/2) 
                 Ty21a (control) 
                 1 mL 
                 4 
                 5 × 10 8   
                 10 5   
               
               
                   
               
            
           
         
       
     
     IV.H. Example 8: Immunogenicity, Serology, Functional Profiling, and Antibody Dependent Enhancement 
     Antibody assessments for NHPs. essentially the same assays for ELISA and neutralization assays as described above for mice are used, but rhesus-specific reagents are used as appropriate. Sera taken prior to immunization and prior to challenge are assessed. However, in addition the Alter laboratory at Ragon Institute will conduct the following assessments of systems serology, function, and antibody dependent enhancement (ADE). 
     Systems Serology. SARS-CoV-2 antigen is carboxy-coupled to Luminex beads. A customized Luminex is used to capture antigen-specific antibody subclass, isotypes, Fc-receptor, lectin-like molecules (DC-Sign, CD23, Dectin), complement initiators (C1q, MBL), innate immune receptors, and lectin binding profiles (SNA: sialic acid, RCA: galactose, AAL: fucose). 
     Functional profiling. Functional assays covering a range of effector mechanisms (ADCC, phagocytosis, complement, antigen uptake) carried out by a diverse set of innate effector cells (NK, dendritic cells, neutrophils, monocytes, macrophages, etc.) are utilized to assess antibody functionality: 
     a) Antibody-dependent phagocytosis is assessed using antigen-coated fluorescent bead uptake by monocytes, monocyte-derived mϕs, neutrophils, or monocyte-derived dendritic cells (DCs) (McAndrew, E. G., et al., J Vis Exp. 57:e3588 (2011)); 
     b) Antibody-dependent NK cell degranulation assesses NK cell activation, a correlate of ADCC, following stimulation with antigen-coated bar-coded beads (Jegaskanda, S., et al., J Immunol 190:1837-48 (2013)); 
     c) Antibody-dependent complement deposition assesses deposition of C3b/C3d onto antigen-coated beads following Ab-co-culture in the presence of guinea pig complement; 
     d) Antibody-dependent cellular cytotoxicity probes Ab-mediated cytotoxicity by innate immune effector cells (NK cells, complement, neutrophils) against antigen-coated NK-resistant CEM cells (Gomez-Roman, V. R., et al.,  J Immunol Methods  308:53-67 (2006)). 
     Antibody dependent enhancement (ADE). ADE likely occurs through multiple mechanisms that include suboptimal neutralization by an antibody followed by antibody bound virus infecting a cell via an Fc receptor. In the Ragon Institute BL3, we will employ a cell-based ADE assay to measure uptake and inflammation following antibody-opsonized virus exposure to THP1 cells, similar assays used to evaluate Dengue-virus driven ADE (Chan, K. R., et al.,  Proc Natl Acad Sci USA  111:2722-7 (2014)). Authentic SARS-CoV-2 virus are co-cultured with virus at multiple MOIs, and then added to the monocyte cell line THP-1 and infection is quantified using anti-SARS CoV-2 antibody staining. 
     IV.I. Example 9: Multiparameter Flow Cytometry and Intracellular Cytokine Staining Assessments of NHPs 
     Multiparameter flow cytometry and in tracellular cytokine staining assessments of NHPs is done as described in previous studies with NHPs with malaria vaccines (Epstein, J. E., et al.,  Science  334: 475-80 (2011); Ishizuka, A. S., et al.,  Nat Med.  22:614-23 (2016)), and has been done for COVID-19 vaccines (Chandrashekar, A., et al.,  Science  369(6505):812-7 (2020)). Focus is on assessing Th1 (e.g. IFN-γ) and Th2 (e.g. IL-4) responses on cryopreserved NHP PBMCs using the same pools of overlapping peptides described in Ishizuka, A. S., et al.,  Nat Med.  22: 614-23 (2016), and assesses cells from the spleen post-sacrifice. Statistical Analysis of flow cytometry studies are done as we have described (Wang S. F., et al.,  Biochem Biophys Res Commun.  451:208-14 (2014); Wu, Y, et al.,  J Infect Dis  215:259-68 (2017)). In brief, data is analyzed using FlowJo v9.8.5 (Tree Star). Statistical analyses are performed with Pestle v1.7 and SPICE v5.3 (M. Roederer) 49, JMP 11 (SAS), Prism 6 (GraphPad), and R v3.2.2 with RStudio v0.99.483. Non-parametric tests are used for association with protection (eg. Kruskal-Wallis with Dunn&#39;s post-test correction for multiple comparisons, Wilcoxon matched-pairs signed rank test with Bonferroni correction for multiple comparisons (Ishizuka A. S., et al.,  Nat Med  22: 614-23 (2016)). 
     IV.J. Example 10: Vaccine Efficacy Assessments in Hamsters and NHPs 
     The following parameters are followed to assess vaccine efficacy in hamsters and NHPs: a) General appearance, weight loss and clinical signs; b) Infectious viral loads and viral RNA in the respiratory tract as determined by testing with oral swabs on days 1, 2, 4 and 7 post challenge in hamsters and NHPs and bronchoalveolar lavage (BAL) of NHPs on days 2, 4, and 7 post-challenge; c) Tissue histopathology, and d) qPCR testing of lungs, nares, GI tract and other tissues after sacrificing the animals. 
     IV.K. Example 11: Vaccine Efficacy Assessments of Additional Recombinant Ty21a Strains Expressing SARS-CoV-2 RBD, M, N, eS, and Combinations Thereof 
     Mice, hamsters, and/or NHPs are immunized with Ty21a-RBD, Ty21a-RBD-AR, Ty21a-M, Ty21a-M-AR, Ty21a-N, Ty21a-N-AR, Ty21a-eS-M-N, Ty21a-eS-M-N-AR, Ty21a-eS+Ty21a-M+Ty21a-N, Ty21a-eS-AR+Ty21a-M-AR+Ty21a-N-AR, or Ty21a and assessed for vaccine efficacy (VE) as described in Examples 6 and 7 with the caveat that M and N proteins and peptide pools will also be used for the antibody and cellular immunology assays. 
     Mouse immunogenicity. Mice are immunized IP as described above (Table 1). The immunological responses in the 10 vaccine candidates and Ty21a control (Table 4) are used to determine which candidates are assessed in hamsters. If AR and non-AR immunogens have similar immunological responses, the AR immunogen is selected. If the Ty21a-eS and Ty21a-RBD immunogens have similar neutralization titers, Ty21a-eS is selected. 
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 11 groups of 10 mice (5M/5F) (total = 110) receive 3 
               
               
                 doses at 2-week intervals. Serum/PBMCs collected pre-immunization 
               
               
                 and 10 days post dose 3. Splenocytes collected 10 days post dose 3. 
               
            
           
           
               
               
               
               
            
               
                 Group 
                 Immunogen 
                 Volume 
                 Dose (CFU) 
               
               
                   
               
               
                 1 
                 Ty21a-RBD 
                 0.2 mL 
                 5 × 10 7   
               
               
                 2 
                 Ty21a-RBD-AR 
                 0.2 mL 
                 5 × 10 7   
               
               
                 3 
                 Ty21a-M 
                 0.2 mL 
                 5 × 10 7   
               
               
                 4 
                 Ty21a-M-AR 
                 0.2 mL 
                 5 × 10 7   
               
               
                 5 
                 Ty21a-N 
                 0.2 mL 
                 5 × 10 7   
               
               
                 6 
                 Ty21a-N-AR 
                 0.2 mL 
                 5 × 10 7   
               
               
                 7 
                 Ty21a-eS-M-N 
                 0.2 mL 
                 5 × 10 7   
               
               
                 8 
                 Ty21a-eS-M-N-AR 
                 0.2 mL 
                 5 × 10 7   
               
               
                  9* 
                 Ty21a-eS + 
                 0.066 mL + 0.066 
                 5.1 × 10 7    
               
               
                   
                 Ty21a-M + 
                 mL + 0.066 mL 
                   
               
               
                   
                 Ty21a-N 
                   
                   
               
               
                 10* 
                 Ty21a-eSAR + 
                 0.066 mL + 0.066 
                 5.1 × 10 7    
               
               
                   
                 Ty21a-M-AR + 
                 mL + 0.066 mL 
                   
               
               
                   
                 Ty21a-N-AR 
                   
                   
               
               
                 11  
                 Ty21a 
                 0.2 mL 
                 5 × 10 7   
               
               
                   
               
               
                 *1.7 × 10 7  CFU of each of the 3 immunogens. 
               
            
           
         
       
     
     Hamster antibody responses and VE studies. Hamsters are immunized as described above in Table 2 and below in Table 5 for the relevant vaccine candidates with Ty21a alone as control. The final down selection of immunogens is based on the results of the mouse immunogenicity studies (Table 4) and the other considerations described above for the assessment of mouse immunogenicity. Antibody responses and VE are assessed as described above in Examples 6-10. 
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 Syrian hamsters are immunized 3 times at 2-day intervals by IN instillation 
               
               
                 of 0.05 mL per naris on days 1, 3, and 5, and challenged by IN inoculation 
               
               
                 of 0.05 mL per naris of SARS-CoV-2 28 days later on day 33. 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 Total Immunizing 
                 Challenge 
               
               
                 Group 
                 N (M/F) 
                 Immunogen 
                 Vol (mL) 
                 Dose (CFU) 
                 Dose (PFU) 
               
               
                   
               
               
                 1 
                 10 (5/5) 
                 Ty21a-eS-M-N-AR 
                 0.1 mL 
                 5 × 10 7   
                 10 5   
               
               
                 2 
                 10 (5/5) 
                 Ty21a-eS-AR + 
                 0.0333 + 
                 5.1 × 10 7   
                 10 5   
               
               
                   
                   
                 Ty21a-M-AR + 
                 0.0333 + 
                 (1.7 × 10 7  + 1.7 × 
                   
               
               
                   
                   
                 Ty21a-N-AR 
                 0.0333 
                 (Ty21a-eS construct) × 10 7 ) 
                   
               
               
                 3 
                 10 (5/5) 
                 Ty21a (control) 
                 0.1 mL 
                 5 × 10 7   
                 10 5   
               
               
                   
               
            
           
         
       
     
     NHP immunogenicity and VE studies. NHPs are immunized as described in Table 3 and below in Table 6 for the relevant vaccine candidates with Ty21a alone as control. The final down selection of immunogens is based on the results of the mouse and hamster immunogenicity studies (Tables 4 and 5) and hamster VE studies, and the other considerations described above for the assessment of mouse immunogenicity. Antibody and cellular immune responses and VE are assessed as described above in Examples 6-10. 
     
       
         
           
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                 NHPs are immunized 4 times at 2-day intervals by IN instillation of 0.5 mL per naris 
               
               
                 on days 1, 3, 5, and 7 (regimen in humans for Ty21a taken orally) and challenged 
               
               
                 by IN inoculation of 0.5 mL per naris of SARS-CoV-2 28 days later on day 35. 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 Total Immunizing 
                 Challenge 
               
               
                 Group 
                 N (M/F) 
                 Immunogen 
                 Vol (mL) 
                 Dose (CFU) 
                 Doe (PFU) 
               
               
                   
               
               
                 1 
                 4 (2/2) 
                 Ty21a-eS-M-N-AR 
                 1 
                 6 × 10 8   
                 10 5   
               
               
                 2 
                 4 (2/2) 
                 Ty21a-eS-AR + 
                 0.0333 + 
                 6 × 10 8   
                 10 5   
               
               
                   
                   
                 Ty21a-M-AR + 
                 0.0333 + 
                 (2 × 10 8  + 2 × 10 8  + 2 × 10 8 ) 
                   
               
               
                   
                   
                 Ty21a-N-AR 
                 0.0333 
                   
                   
               
               
                 3 
                 4 (2/2) 
                 Ty21a (control) 
                 1 
                 6 × 10 8   
                 10 5   
               
               
                   
               
            
           
         
       
     
     IV.L. Example 12: Chromosomal Integration of eS Gene Fragment 
     A synthesized sequence of eS fused to the secretion signal Hly (SEQ ID NO: 5;  FIG. 9 ) (Thomas, S., et al.,  Biochim Biophys Acta.  1843:1629-41 (2014)) was codon optimized for expression in  Salmonella  and introduced into the shuttle plasmid (pUC57). The eS Hlya cassette was PCR amplified and inserted into plasmid pMDTV-Lpp-PA Hlys after removal of the PA sequence fragment. The resultant plasmid pMDTV-S-ecto was used to transform competent TY21a. The eS fusion protein was strongly expressed in both the pellet ( FIG. 7A ) as well as in the supernatant ( FIG. 7B ) of transformed Ty21a containing plasmid Ty21a+pTV-Lpp-S-ecto 9 Hlys (Kan). The results shown in  FIGS. 7A-7B  demonstrate that Ty21a are capable of efficiently expressing the eS fusion protein. 
     Next, the gene fragment encoding the eS fusion protein was integrated into the chromosome of Ty21a using k-red recombineering technology (Datsenko K. A., and B. L. Wanner,  Proc Natl Acad Sci USA  97:6640-5 (2000); Dharmasena M. N., et al.,  Int J Med Microbiol.  303:105-13 (2013)) and previously described methods (Wu, Y, et al.,  J Infect Dis  215:259-68 (2017); Sim, B. K. L., et al.,  NPJ Vaccines  2:17 (2017)). The constitutive lpp promoter (Lpp-outer membrane lipoprotein) (Inouye S., and M. Inouye,  Nucleic Acids Res  13:3101-10 (1985)) was used to ensure high expression of the eS fusion protein from the integrated gene fragment. The gene fragment encoding the eS fusion protein was inserted between tviD and vexA in the chromosome of Ty21a ( FIG. 5B ), resulting in a Ty21a-S-ecto clone. 
     To show that the chromosomally integrated eS gene fragment expressed eS protein in both the pellet and supernatant of the Ty21a-S-ecto clone, a culture of clone Ty21a-S-ecto at OD 0.15 was centrifuged, pelleted, and mixed with sample buffer. Sample buffer was then subjected to electrophoresis on a 4-20% Tris Glycine gel and blotted onto a PVDF membrane. The PVDF membrane was probed with anti-SARS-CoV-2/2019-nCoV Spike polyclonal rabbit antibody (1:1500 dilution) ( FIG. 8A ). Similarly, supernatant of an OD 0.5 culture of clone Ty21a-S-ecto was mixed with sample buffer and loaded onto a 4-20% Tris Glycine gel, subjected to electrophoresis, blotted onto a PVDF membrane, and probed with anti-SARS-CoV-2/2019-nCoV Spike polyclonal rabbit antibody (1:1500 dilution) ( FIG. 8B ). The results shown in  FIGS. 8A-8B  demonstrate that clone Ty21a-S-ecto, which has the eS gene fragment integrated in the chromosome of Ty21a, is capable of efficiently expressing the eS fusion protein 
     IV.M. Example 13: Chromosomal Integration of RBD-HlyA 
     The receptor binding domain (RBD; SEQ ID NO: 11) of the SARS-CoV2 is at position 319 to 541 on the amino acid sequence of the spike protein. The RBD was PCR-amplified from the pUC57 plasmid carrying the entire eS (See Example 1). The RBD was then used to replace the eS gene on the pMDTV-LPP-eS-HlyA+AR plasmid, as follows: The pMDTV-LPP-eS-HlyA+AR plasmid was linearized by PCR, excluding the eS gene. The linearized plasmid without the eS gene was then assembled to the RBD using NEBuilder® HiFi DNA Assembly (New England Bioloabs #E5520S), to generate pMDTV-LPP-RBD-HlyA+AR. The construct was confirmed by PCR and sequencing. The RBD was fused to the HlyA secretion signal (RBD-HlyA). Next, the RBD-HlyA and the  Shigella sonnei  acid resistance cassette were integrated to the Ty21a genome at the tviE locus as follows: The pMDTV-LPP-RBD-HlyA-AR was used as a template for a PCR, amplifying from the 5′ end of the Tvid to the HlyA. The amplicon (AR-LPP-RBD-HlyA) was used to replace the eS-HlyA in the transgenic Ty21a-eS construct (described in Example 12) by homologous recombination, using the i-red recombineering technology, as was described in Example 1. In this case, the HlyA secretion signal was already present in the transgenic bacteria and was used as the right homologous arm, while the 3′ end of the tviD gene was used as the left homologous arm. The kanamycin resistance gene was used as a selection marker as it was removed in the host Ty21a-eS transgenic strain. A schematic showing the construction of the Ty21a-RBD-AR construct is shown in  FIG. 10 . 
     Additionally, PCR reactions were used to confirm full integration of the AR-LPP-RBD-HlyA construct ( FIG. 11A-11C ). The inserted fragment was PCR-amplified using primer 27 and primer 107, flanking the homologous arms on both 5′ and 3′ ends ( FIG. 11A ). The expected amplicon sizes were 3.3 Kb for the WT, 9.7 Kb for the background Ty21-eS construct, and 13.3 Kb for the transgenic Ty21-RBD+AR. Out of the 8 clones tested, only clone 3 was positive for the insertion. 
     Conformation of the integration on the 5′ end was done by PCR using primer 27, upstream to the left homology arm and primer 37, inside the inserted fragment, downstream to the kanR gene ( FIG. 11B ). The expected amplicon sizes were 0.7 Kb for the background Ty21-eS construct and the Ty21 eS+AR construct, and 2.1 Kb for the transgenic Ty21-RBD+AR. No amplification is expected on WT. Clone 3 was positive for the insertion while clone 1 was not. 
     Conformation of the integration on the 3′ end was done by PCR using primer 64, downstream to the right homology arm (HlyA), on the HlyB gene and primer 89, inside the inserted fragment, on the 5′ of the RBD ( FIG. 11C ). The expected amplicon sizes were 4.1 Kb for the background Ty21-eS construct and the Ty21 eS+AR construct, and 1.1 Kb for the transgenic Ty21-RBD+AR. No amplification was expected for WT. Both clones were positive for the insertion on the 3′. The PCR reactions confirmed full integration for clone 3 and only partial, non-complete integration for other clones. 
     It is to be appreciated that the Detailed Description section, and not the Summary and Abstract sections, is intended to be used to interpret the claims. The Summary and Abstract sections may set forth one or more but not all exemplary aspects or embodiments of the present invention as contemplated by the inventor(s), and thus, are not intended to limit the present invention and the appended claims in any way. 
     The present invention has been described above with the aid of functional building blocks illustrating the implementation of specified functions and relationships thereof. The boundaries of these functional building blocks have been arbitrarily defined herein for the convenience of the description. Alternate boundaries can be defined so long as the specified functions and relationships thereof are appropriately performed. 
     The foregoing description of the specific aspects or embodiments will so fully reveal the general nature of the invention that others can, by applying knowledge within the skill of the art, readily modify and/or adapt for various applications such specific aspects or embodiments, without undue experimentation, without departing from the general concept of the present invention. Therefore, such adaptations and modifications are intended to be within the meaning and range of equivalents of the disclosed aspects or embodiments, based on the teaching and guidance presented herein. It is to be understood that the phraseology or terminology herein is for the purpose of description and not of limitation, such that the terminology or phraseology of the present specification is to be interpreted by the skilled artisan in light of the teachings and guidance. 
     The breadth and scope of the present invention should not be limited by any of the above-described exemplary aspects or embodiments, but should be defined only in accordance with the following claims and their equivalents. 
     
       
         
           
               
             
               
                 TABLE 7 
               
               
                   
               
               
                 Amino Acid and Nucleic Acid Sequences 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
            
               
                 SEQ ID NO: 1 (Modified 
                 MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHS 
               
               
                 SARS CoV-2 eS) 
                 TQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNI 
               
               
                   
                 IRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNK 
               
               
                   
                 SWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGY 
               
               
                   
                 FKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLT 
               
               
                   
                 PGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETK 
               
               
                   
                 CTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASV 
               
               
                   
                 YAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSF 
               
               
                   
                 VIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYN 
               
               
                   
                 YLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPT 
               
               
                   
                 NGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTG 
               
               
                   
                 VLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITP 
               
               
                   
                 GTNTSNQVAVLYQGVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCL 
               
               
                   
                 IGAEHVNNSYECDIPIGAGICASYQTQTNSPGSASSVASQSIIAYTMSLG 
               
               
                   
                 AENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECS 
               
               
                   
                 NLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGF 
               
               
                   
                 NFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLI 
               
               
                   
                 CAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAM 
               
               
                   
                 QMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQD 
               
               
                   
                 VVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDPPEAEVQIDRLITGR 
               
               
                   
                 LQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLM 
               
               
                   
                 SFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGT 
               
               
                   
                 HWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKE 
               
               
                   
                 ELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDL 
               
               
                   
                 QELGKYEQYIKWPWYIWLGFIAGLIAIVMVTIMLCCMTSCCSCLKGCCSC 
               
               
                   
                 GSCCKFDEDDSEPVLKGVKLHYTGYIPEAPRDGQAYVRKDGEWVLLSTFL 
               
               
                   
               
               
                 SEQ ID NO: 2 (SARS- 
                 MADSNGTITVEELKKLLEQWNLVIGFLFLTWICLLQFAYANRNRFLYIIK 
               
               
                 CoV2 M Protein) 
                 LIFLWLLWPVTLACFVLAAVYRINWITGGIAIAMACLVGLMWLSYFIASF 
               
               
                   
                 RLFARTRSMWSFNPETNILLNVPLHGTILTRPLLESELVIGAVILRGHLR 
               
               
                   
                 IAGHHLGRCDIKDLPKEITVATSRTLSYYKLGASQRVAGDSGFAAYSRYR 
               
               
                   
                 IGNYKLNTDHSSSSDNIALLVQ 
               
               
                   
               
               
                 SEQ ID NO: 3 (SARS- 
                 MSDNGPQNQRNAPRITFGGPSDSTGSNQNGERSGARSKQRRPQGLPNNTA 
               
               
                 CoV2 N Protein) 
                 SWFTALTQHGKEDLKFPRGQGVPINTNSSPDDQIGYYRRATRRIRGGDGK 
               
               
                   
                 MKDLSPRWYFYYLGTGPEAGLPYGANKDGIIWVATEGALNTPKDHIGTRN 
               
               
                   
                 PANNAAIVLQLPQGTTLPKGFYAEGSRGGSQASSRSSSRSRNSSRNSTPG 
               
               
                   
                 SSRGTSPARMAGNGGDAALALLLLDRLNQLESKMSGKGQQQQGQTVTKKS 
               
               
                   
                 AAEASKKPRQKRTATKAYNVTQAFGRRGPEQTQGNFGDQELIRQGTDYKH 
               
               
                   
                 WPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLDDKDPNFKDQV 
               
               
                   
                 ILLNKHIDAYKTFPPTEPKKDKKKKADETQALPQRQKKQQTVTLLPAADL 
               
               
                   
                 DDFSKQLQQSMSSADSTQA 
               
               
                   
               
               
                 SEQ ID NO: 4 (Amino 
                 MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHS 
               
               
                 Acid Sequence at 
                 TQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNI 
               
               
                 Positions 1-1296 of 
                 IRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNK 
               
               
                 SARS CoV-2 S Protein) 
                 SWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGY 
               
               
                   
                 FKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLT 
               
               
                   
                 PGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETK 
               
               
                   
                 CTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASV 
               
               
                   
                 YAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSF 
               
               
                   
                 VIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYN 
               
               
                   
                 YLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPT 
               
               
                   
                 NGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTG 
               
               
                   
                 VLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITP 
               
               
                   
                 GTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCL 
               
               
                   
                 IGAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVASQSIIAYTMSLG 
               
               
                   
                 AENSVAYSNNSIAIPTNFTI 
               
               
                   
               
               
                 SEQ ID NO: 5 (Modified 
                 MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHS 
               
               
                 SARS CoV-2 eS) 
                 TQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNI 
               
               
                   
                 IRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNK 
               
               
                   
                 SWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGY 
               
               
                   
                 FKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLT 
               
               
                   
                 PGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETK 
               
               
                   
                 CTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASV 
               
               
                   
                 YAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSF 
               
               
                   
                 VIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYN 
               
               
                   
                 YLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPT 
               
               
                   
                 NGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTG 
               
               
                   
                 VLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITP 
               
               
                   
                 GTNTSNQVAVLYQGVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCL 
               
               
                   
                 IGAEHVNNSYECDIPIGAGICASYQTQTNSPGSASSVASQSIIAYTMSLG 
               
               
                   
                 AENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECS 
               
               
                   
                 NLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGF 
               
               
                   
                 NFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLI 
               
               
                   
                 CAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAM 
               
               
                   
                 QMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQD 
               
               
                   
                 VVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDPPEAEVQIDRLITGR 
               
               
                   
                 LQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLM 
               
               
                   
                 SFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGT 
               
               
                   
                 HWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKE 
               
               
                   
                 ELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDL 
               
               
                   
                 QELGKYEQGYIPEAPRDGQAYVRKDGEWVLLSTFL 
               
               
                   
                 ALAYGSQGNLNPLINEISKIISAAGNFDVKEERAAASLLQLSGNASDFSY 
               
               
                   
                 GRNSITLTASA 
               
               
                   
               
               
                 SEQ ID NO: 6(Ty21a-eS 
                 cccgttcctcattgatttgattgctaacgtcatgagcattactctatcat 
               
               
                 Construct) 
                 tccagaatgcctcaatgaacaagttgtttgagaaagagtgtcgcaatgtt 
               
               
                   
                 gcaaccagagcccttaaatatgtacgccagaagaaaactgaagggcgtct 
               
               
                   
                 ggatgaagcattgtctgtattgattagcctgaaacgaattgagcctgatg 
               
               
                   
                 tttctcgtctgatgcgtgaatataagcaaattatcagattatttaatgag 
               
               
                   
                 tcacggaaggatggcggtagcactatcacgtcttatgaacatctagacta 
               
               
                   
                 tgcgaaaaaattactcgtttttgatagcgaaaatgcctatgccttgaaat 
               
               
                   
                 atgccgcattaaatgcaatgcatttacgcgactacacgcaggctttgcag 
               
               
                   
                 tattggcagcgactggagaaagtgaatggaccaacggagccggtgacaag 
               
               
                   
                 gcagatctcgacctgcataaccgcattacaaaaaaatacatcagggaagt 
               
               
                   
                 cgtaacccggggtgtaggctggagctgcttcgaagttcctatactttcta 
               
               
                   
                 gagaataggaacttcggaataggaacttcaagatcccctcacgctgccgc 
               
               
                   
                 aagcactcagggcgcaagggctgctaaaggaagcggaacacgtagaaagc 
               
               
                   
                 cagtccgcagaaacggtgctgaccccggatgaatgtcagctactgggcta 
               
               
                   
                 tctggacaagggaaaacgcaagcgcaaagagaaagcaggtagcttgcagt 
               
               
                   
                 gggcttacatggcgatagctagactgggcggttttatggacagcaagcga 
               
               
                   
                 accggaattgccagctggggcgccctctggtaaggttgggaagccctgca 
               
               
                   
                 aagtaaactggatggctttcttgccgccaaggatctgatggcgcagggga 
               
               
                   
                 tcaagatctgatcaagagacaggatgaggatcgtttcgcatgattgaaca 
               
               
                   
                 agatggattgcacgcaggttctccggccgcttgggtggagaggctattcg 
               
               
                   
                 gctatgactgggcacaacagacaatcggctgctctgatgccgccgtgttc 
               
               
                   
                 cggctgtcagcgcaggggcgcccggttctttttgtcaagaccgacctgtc 
               
               
                   
                 cggtgccctgaatgaactgcaggacgaggcagcgcggctatcgtggctgg 
               
               
                   
                 ccacgacgggcgttccttgcgcagctgtgctcgacgttgtcactgaagcg 
               
               
                   
                 ggaagggactggctgctattgggcgaagtgccggggcaggatctcctgtc 
               
               
                   
                 atctcaccttgctcctgccgagaaagtatccatcatggctgatgcaatgc 
               
               
                   
                 ggcggctgcatacgcttgatccggctacctgcccattcgaccaccaagcg 
               
               
                   
                 aaacatcgcatcgagcgagcacgtactcggatggaagccggtcttgtcga 
               
               
                   
                 tcaggatgatctggacgaagagcatcaggggctcgcgccagccgaactgt 
               
               
                   
                 tcgccaggctcaaggcgcgcatgcccgacggcgaggatctcgtcgtgacc 
               
               
                   
                 catggcgatgcctgcttgccgaatatcatggtggaaaatggccgcttttc 
               
               
                   
                 tggattcatcgactgtggccggctgggtgtggcggaccgctatcaggaca 
               
               
                   
                 tagcgttggctacccgtgatattgctgaagagcttggcggcgaatgggct 
               
               
                   
                 gaccgcttcctcgtgctttacggtatcgccgctcccgattcgcagcgcat 
               
               
                   
                 cgccttctatcgccttcttgacgagttcttctgagcgggactctggggtt 
               
               
                   
                 cgaaatgaccgaccaagcgacgcccaacctgccatcacgagatttcgatt 
               
               
                   
                 ccaccgccgccttctatgaaaggttgggcttcggaatcgttttccgggac 
               
               
                   
                 gccggctggatgatcctccagcgcggggatctcatgctggagttcttcgc 
               
               
                   
                 ccaccccagcttcaaaagcgctctgaagttcctatactttctagagaata 
               
               
                   
                 ggaacttcggaataggaactaaggaggatattcatatatgcattaatgtc 
               
               
                   
                 taacaattcgttcaagccgacgccgcttcgcggcgcggcttaactcaagc 
               
               
                   
                 gttagatgcactaagcacataattgctcacagccaaactatcaggtcaag 
               
               
                   
                 tctgcttttattatttttaagcgtgcataataagccctacacaaattggg 
               
               
                   
                 agatatatcatgaaaggctggctttttcttgttatcgcaatagttggcga 
               
               
                   
                 agtaatcgcaacatccgcattaaaatctagcgagggctttactgtcgacc 
               
               
                   
                 aaaaaaatattgacaacataaaaaactttgtgttatacttgtaacgGact 
               
               
                   
                 ctagagggtattaataatgaaaggagaggacatGAGCTCATGTTTGTGTT 
               
               
                   
                 TCTGGTGCTGCTGCCGCTGGTGAGCAGCCAGTGCGTGAACCTGACCACCC 
               
               
                   
                 GCACCCAGCTGCCGCCGGCGTATACCAACAGCTTTACCCGCGGCGTGTAT 
               
               
                   
                 TATCCGGATAAAGTGTTTCGCAGCAGCGTGCTGCATAGCACCCAGGATCT 
               
               
                   
                 GTTTCTGCCGTTTTTTAGCAACGTGACCTGGTTTCATGCGATTCATGTGA 
               
               
                   
                 GCGGCACCAACGGCACCAAACGCTTTGATAACCCGGTGCTGCCGTTTAAC 
               
               
                   
                 GATGGCGTGTATTTTGCGAGCACCGAAAAAAGCAACATTATTCGCGGCTG 
               
               
                   
                 GATTTTTGGCACCACCCTGGATAGCAAAACCCAGAGCCTGCTGATTGTGA 
               
               
                   
                 ACAACGCGACCAACGTGGTGATTAAAGTGTGCGAATTTCAGTTTTGCAAC 
               
               
                   
                 GATCCGTTTCTGGGCGTGTATTATCATAAAAACAACAAAAGCTGGATGGA 
               
               
                   
                 AAGCGAATTTCGCGTGTATAGCAGCGCGAACAACTGCACCTTTGAATATG 
               
               
                   
                 TGAGCCAGCCGTTTCTGATGGATCTGGAAGGCAAACAGGGCAACTTTAAA 
               
               
                   
                 AACCTGCGCGAATTTGTGTTTAAAAACATTGATGGCTATTTTAAAATTTA 
               
               
                   
                 TAGCAAACATACCCCGATTAACCTGGTGCGCGATCTGCCGCAGGGCTTTA 
               
               
                   
                 GCGCGCTGGAACCGCTGGTGGATCTGCCGATTGGCATTAACATTACCCGC 
               
               
                   
                 TTTCAGACCCTGCTGGCGCTGCATCGCAGCTATCTGACCCCGGGCGATAG 
               
               
                   
                 CAGCAGCGGCTGGACCGCGGGCGCGGCGGCGTATTATGTGGGCTATCTGC 
               
               
                   
                 AGCCGCGCACCTTTCTGCTGAAATATAACGAAAACGGCACCATTACCGAT 
               
               
                   
                 GCGGTGGATTGCGCGCTGGATCCGCTGAGCGAAACCAAATGCACCCTGAA 
               
               
                   
                 AAGCTTTACCGTGGAAAAAGGCATTTATCAGACCAGCAACTTTCGCGTGC 
               
               
                   
                 AGCCGACCGAAAGCATTGTGCGCTTTCCGAACATTACCAACCTGTGCCCG 
               
               
                   
                 TTTGGCGAAGTGTTTAACGCGACCCGCTTTGCGAGCGTGTATGCGTGGAA 
               
               
                   
                 CCGCAAACGCATTAGCAACTGCGTGGCGGATTATAGCGTGCTGTATAACA 
               
               
                   
                 GCGCGAGCTTTAGCACCTTTAAATGCTATGGCGTGAGCCCGACCAAACTG 
               
               
                   
                 AACGATCTGTGCTTTACCAACGTGTATGCGGATAGCTTTGTGATTCGCGG 
               
               
                   
                 CGATGAAGTGCGCCAGATTGCGCCGGGCCAGACCGGCAAAATTGCGGATT 
               
               
                   
                 ATAACTATAAACTGCCGGATGATTTTACCGGCTGCGTGATTGCGTGGAAC 
               
               
                   
                 AGCAACAACCTGGATAGCAAAGTGGGCGGCAACTATAACTATCTGTATCG 
               
               
                   
                 CCTGTTTCGCAAAAGCAACCTGAAACCGTTTGAACGCGATATTAGCACCG 
               
               
                   
                 AAATTTATCAGGCGGGCAGCACCCCGTGCAACGGCGTGGAAGGCTTTAAC 
               
               
                   
                 TGCTATTTTCCGCTGCAGAGCTATGGCTTTCAGCCGACCAACGGCGTGGG 
               
               
                   
                 CTATCAGCCGTATCGCGTGGTGGTGCTGAGCTTTGAACTGCTGCATGCGC 
               
               
                   
                 CGGCGACCGTGTGCGGCCCGAAAAAAAGCACCAACCTGGTGAAAAACAAA 
               
               
                   
                 TGCGTGAACTTTAACTTTAACGGCCTGACCGGCACCGGCGTGCTGACCGA 
               
               
                   
                 AAGCAACAAAAAATTTCTGCCGTTTCAGCAGTTTGGCCGCGATATTGCGG 
               
               
                   
                 ATACCACCGATGCGGTGCGCGATCCGCAGACCCTGGAAATTCTGGATATT 
               
               
                   
                 ACCCCGTGCAGCTTTGGCGGCGTGAGCGTGATTACCCCGGGCACCAACAC 
               
               
                   
                 CAGCAACCAGGTGGCGGTGCTGTATCAGGGCGTGAACTGCACCGAAGTGC 
               
               
                   
                 CGGTGGCGATTCATGCGGATCAGCTGACCCCGACCTGGCGCGTGTATAGC 
               
               
                   
                 ACCGGCAGCAACGTGTTTCAGACCCGCGCGGGCTGCCTGATTGGCGCGGA 
               
               
                   
                 ACATGTGAACAACAGCTATGAATGCGATATTCCGATTGGCGCGGGCATTT 
               
               
                   
                 GCGCGAGCTATCAGACCCAGACCAACAGCCCGGGCAGCGCGAGCAGCGTG 
               
               
                   
                 GCGAGCCAGAGCATTATTGCGTATACCATGAGCCTGGGCGCGGAAAACAG 
               
               
                   
                 CGTGGCGTATAGCAACAACAGCATTGCGATTCCGACCAACTTTACCATTA 
               
               
                   
                 GCGTGACCACCGAAATTCTGCCGGTGAGCATGACCAAAACCAGCGTGGAT 
               
               
                   
                 TGCACCATGTATATTTGCGGCGATAGCACCGAATGCAGCAACCTGCTGCT 
               
               
                   
                 GCAGTATGGCAGCTTTTGCACCCAGCTGAACCGCGCGCTGACCGGCATTG 
               
               
                   
                 CGGTGGAACAGGATAAAAACACCCAGGAAGTGTTTGCGCAGGTGAAACAG 
               
               
                   
                 ATTTATAAAACCCCGCCGATTAAAGATTTTGGCGGCTTTAACTTTAGCCA 
               
               
                   
                 GATTCTGCCGGATCCGAGCAAACCGAGCAAACGCAGCTTTATTGAAGATC 
               
               
                   
                 TGCTGTTTAACAAAGTGACCCTGGCGGATGCGGGCTTTATTAAACAGTAT 
               
               
                   
                 GGCGATTGCCTGGGCGATATTGCGGCGCGCGATCTGATTTGCGCGCAGAA 
               
               
                   
                 ATTTAACGGCCTGACCGTGCTGCCGCCGCTGCTGACCGATGAAATGATTG 
               
               
                   
                 CGCAGTATACCAGCGCGCTGCTGGCGGGCACCATTACCAGCGGCTGGACC 
               
               
                   
                 TTTGGCGCGGGCGCGGCGCTGCAGATTCCGTTTGCGATGCAGATGGCGTA 
               
               
                   
                 TCGCTTTAACGGCATTGGCGTGACCCAGAACGTGCTGTATGAAAACCAGA 
               
               
                   
                 AACTGATTGCGAACCAGTTTAACAGCGCGATTGGCAAAATTCAGGATAGC 
               
               
                   
                 CTGAGCAGCACCGCGAGCGCGCTGGGCAAACTGCAGGATGTGGTGAACCA 
               
               
                   
                 GAACGCGCAGGCGCTGAACACCCTGGTGAAACAGCTGAGCAGCAACTTTG 
               
               
                   
                 GCGCGATTAGCAGCGTGCTGAACGATATTCTGAGCCGCCTGGATCCGCCG 
               
               
                   
                 GAAGCGGAAGTGCAGATTGATCGCCTGATTACCGGCCGCCTGCAGAGCCT 
               
               
                   
                 GCAGACCTATGTGACCCAGCAGCTGATTCGCGCGGCGGAAATTCGCGCGA 
               
               
                   
                 GCGCGAACCTGGCGGCGACCAAAATGAGCGAATGCGTGCTGGGCCAGAGC 
               
               
                   
                 AAACGCGTGGATTTTTGCGGCAAAGGCTATCATCTGATGAGCTTTCCGCA 
               
               
                   
                 GAGCGCGCCGCATGGCGTGGTGTTTCTGCATGTGACCTATGTGCCGGCGC 
               
               
                   
                 AGGAAAAAAACTTTACCACCGCGCCGGCGATTTGCCATGATGGCAAAGCG 
               
               
                   
                 CATTTTCCGCGCGAAGGCGTGTTTGTGAGCAACGGCACCCATTGGTTTGT 
               
               
                   
                 GACCCAGCGCAACTTTTATGAACCGCAGATTATTACCACCGATAACACCT 
               
               
                   
                 TTGTGAGCGGCAACTGCGATGTGGTGATTGGCATTGTGAACAACACCGTG 
               
               
                   
                 TATGATCCGCTGCAGCCGGAACTGGATAGCTTTAAAGAAGAACTGGATAA 
               
               
                   
                 ATATTTTAAAAACCATACCAGCCCGGATGTGGATCTGGGCGATATTAGCG 
               
               
                   
                 GCATTAACGCGAGCGTGGTGAACATTCAGAAAGAAATTGATCGCCTGAAC 
               
               
                   
                 GAAGTGGCGAAAAACCTGAACGAAAGCCTGATTGATCTGCAGGAACTGGG 
               
               
                   
                 CAAATATGAACAGGGCTATATTCCGGAAGCGCCGCGCGATGGCCAGGCGT 
               
               
                   
                 ATGTGCGCAAAGATGGCGAATGGGTGCTGCTGAGCACCTTTCTGGCATTA 
               
               
                   
                 GCCTATGGAAGTCAGGGTAATCTTAATCCATTAATTAATGAAATCAGCAA 
               
               
                   
                 AATCATTTCAGCTGCAGGTAATTTTGATGTTAAAGAGGAAAGAGCGGCCG 
               
               
                   
                 CGTCTTTATTGCAGTTGTCCGGTAATGCCAGTGATTTTTCATATGGACGG 
               
               
                   
                 AACTCAATAACTTTGACAGCATCAGCATAAGAGCTCtttattaatttaaa 
               
               
                   
                 taatagcaatcttactgggctgtgccacataagattgctatttttttgga 
               
               
                   
                 gtcataatggattcttgtcataaaattgattatgggttatacgccctgga 
               
               
                   
                 gattttagcccaataccataacgtctctgttaacccggaagaaattaaac 
               
               
                   
                 atagatttgacacagacgggactggtctgggattaacgtcatggttgctt 
               
               
                   
                 gctgcgaaatctttagaactaaaggtaaaacaggtaaaaaaaacaattga 
               
               
                   
                 ccgattaaactttatttctctgcccgcattagtctggagagaggatggat 
               
               
                   
                 gtcattttattctgactaaagtcagtaaagaagcaaacagatatcttatt 
               
               
                   
                 tttgatctggagcagcgaaatccccgtgttctcgaacagtctgagtttga 
               
               
                   
                 ggcgttatatcaggggcatattattcttattgcttcccgttcttctgtta 
               
               
                   
                 ccgggaaactggcaaaatttgactttacctggtttattcctgccattata 
               
               
                   
                 aaatacaggaaaatatttattgaaacccttgttgtatctgtttttttaca 
               
               
                   
                 attatttgcattaataaccccccttttttttcaggtggttatggacaaag 
               
               
                   
                 tattagtacacagggggttttcaacccttaatgttattactgtcgcatta 
               
               
                   
                 tctgttgtggtggtgtttgagattatactcagcggtttaagaacttacat 
               
               
                   
                 ttttgcacatagtacaagtcggattgatgttgagttgggtgccaaactct 
               
               
                   
                 tccggcatttactggcgctaccgatctcttattttgagagtcgtcgtgtt 
               
               
                   
                 ggtgatactgttgccagggtaagagaattagaccagatccgtaatttcct 
               
               
                   
                 gacaggacaggcattaacatctgttctggacttattattttcattcatat 
               
               
                   
                 tttttgcggtaatgtggtattacagcccaaagcttactctggtgatctta 
               
               
                   
                 ttttcgctgccctgttatgctgcatggtctgtttttattagccccatttt 
               
               
                   
                 gcgacgtcgccttgatgataagttttcacggaatgcggataatcaatctt 
               
               
                   
                 tcctggtggaatcagtcacggcgattaacactataaaagctatggcagtc 
               
               
                   
                 tcacctcagatgacgaacatatgggacaaacaattggcaggatatgttgc 
               
               
                   
                 tgcaggctttaaagtgacagtattagccaccattggtcaacaaggaatac 
               
               
                   
                 agttaatacaaaagactgttatgatcatcaacctgtggttgggtgcacac 
               
               
                   
                 ctggttatttccggggatttaagtattggtcagttaattgcttttaatat 
               
               
                   
                 gcttgcaggtcagattgttgcaccggttattcgccttgcacaaatctggc 
               
               
                   
                 aggatttccagcaggttggtatatcagttacccgccttggtgatgtgctt 
               
               
                   
                 aactctccaactgaaagttatcatgggaaactggcattaccggaaattaa 
               
               
                   
                 tggtgatatcacttttcgtaatatccggtttcgctataagcctgactctc 
               
               
                   
                 cggttattttagataatatcaatctcagtattaagcagggggaggttatt 
               
               
                   
                 ggtattgtcggacgttctggttcaggaaaaagcacattaactaaattaat 
               
               
                   
                 tcaacgtttttatattcctgaaaatggccaggtcttaattgatggacatg 
               
               
                   
                 atcttgcgttggccgatcctaactggttacgtcgtcaggtgggggttgtg 
               
               
                   
                 ttgcaggacaatgtgctgcttaatcgcagtattattgataatatttcact 
               
               
                   
                 ggctaatcctggcatgtccgtcgaaaaagttatttatgcagcgaaattag 
               
               
                   
                 caggtgctcatgattttatttctgaattgcgtgaggggtataacaccatt 
               
               
                   
                 gtcggggaacagggggcaggattatccggaggtcaacgtcaacgcatcgc 
               
               
                   
                 aattgcaagggcgctggtgaacaaccctaaaatactcatctttgatgaag 
               
               
                   
                 caaccagtgctctggattatgagtcggagcatgtcatcatgcgcaatatg 
               
               
                   
                 cacaaaatatgtaagggcagaacggttataatcattgctcatcgtctgtc 
               
               
                   
                 tacagtaaaaaatgcagaccgcattattgtcatggaaaaagggaaaattg 
               
               
                   
                 ttgaacagggtaaacataaggagctgctttctgaaccggaaagtttatac 
               
               
                   
                 agttacttatatcagttacagtcagactaacagaaagaacagaagaatat 
               
               
                   
                 gaaaacatggttaatggggttcagcgagttcctgttgcgctataaacttg 
               
               
                   
                 tctggagtgaaacatggaaaatccggaagcagttagatactccggtacgt 
               
               
                   
                 gaaaaggacgaaaatgaattcttacccgctcatctggaattaattgaaac 
               
               
                   
                 gccggtatccagacggccgcgtctggttgcttattttattatggggtttc 
               
               
                   
                 tggttattgctttcattttatctgttttaggccaggtggaaattgttgcc 
               
               
                   
                 actgcaaatgggaaattaacactcagtgggcgtagcaaagaaattaaacc 
               
               
                   
                 tattgaaaactcgatagttaaagaaattatcgtaaaagaaggagagtcag 
               
               
                   
                 tccggaaaggggatgtgttattaaagcttacagcgctgggagctgaagct 
               
               
                   
                 gatacgttaaaaacgcagtcatcactgttacaggccaggctggaacaaat 
               
               
                   
                 tcggtatcaaattctgagccggtcaattgaattaaataaacttcctgaac 
               
               
                   
                 tgaaacttcctgatgagccttattttcagaatgtatctgaagaggaagta 
               
               
                   
                 ctgcgtttaacttctttgataaaagaacagttttccacatggcaaaatca 
               
               
                   
                 gaagtatcaaaaagaactgaatctggataagaaaagagcagagcgattaa 
               
               
                   
                 caatacttgcccgtataaaccgttatgaaaatgtatcgagggttgaaaaa 
               
               
                   
                 agccgtctggatgatttcaggagcctgttgcataaacaggcaattgcaaa 
               
               
                   
                 acatgctgtacttgagcaggagaataaatatgttgaggcagcaaatgaat 
               
               
                   
                 tacgggtttataaatcgcaactggagcaaattgagagtgagatattgtct 
               
               
                   
                 gcaaaagaagaatatcagcttgtcacgcagctttttaaaaatgaaatttt 
               
               
                   
                 agacaagctaagacaaacaacagacagcattgagttattaactctggagt 
               
               
                   
                 tagagaaaaatgaagagcgtcaacaggcttcagtaatcagggcccctgtt 
               
               
                   
                 tcgggaaaagttcagcaactgaaggttcatactgaaggtggggttgttac 
               
               
                   
                 aacagcggaaacactgatggtcatcgttccggaagatgacacgctggagg 
               
               
                   
                 ttactgctctggtacaaaataaagatattggttttattaacgtcgggcag 
               
               
                   
                 aatgccatcattaaagtggaggcttttccttacacccgatatggttatct 
               
               
                   
                 ggtgggtaaggtaaaaaatataaatttagatgcaatagaagaccagaaac 
               
               
                   
                 tgggactcgtttttaatgtcattgtttctgttgaagagaatgatttgtca 
               
               
                   
                 accgggaataagcacattccattaagctcgggtatggctgtcactgcaga 
               
               
                   
                 aataaagactggaatgcgaagcgtaatcagttatcttcttagtcccctgg 
               
               
                   
                 aagaatctgtaacagaaagtttacatgagcgttaagtctcagagcagcgg 
               
               
                   
                 tatccggctcatatcttctcctgtcagatccaagggcgaattccagcaca 
               
               
                   
                 ctggcggccgttactagtggatccaccggtctcgaggggcgcgccggtac 
               
               
                   
                 cgaattctaattaatgggcatcatttttcagctatttcatttataaaata 
               
               
                   
                 agttatgaaaaaaatcatcatattactaacgacatttttcctgctttcgg 
               
               
                   
                 gatgcactattcccagggcggtatttaaatccagccttattaatcaggac 
               
               
                   
                 gatcctcgttataatctggtcgaagtcacgccgacattaaaactaagcgc 
               
               
                   
                 tcccgatactgtgccgaaaactattgtcgatccggtttttgccgcaaata 
               
               
                   
                 actggcactggacatctttggctaaaggcgatgtgctgcatatcactatt 
               
               
                   
                 ttatcctcgggcggggctggatatttatccaataacgcgagcggcgaccg 
               
               
                   
                 tgcggattttgaaaatattcttgtgactgacagtaataccgttcaggtgc 
               
               
                   
                 cttatgccgggacaatcccggtttctggattggatgtgacgcaactggct 
               
               
                   
                 gatgagatcaaaaagcgactttcgcgcgttgtcctgaatcctcaggtgat 
               
               
                   
                 tgtgacacttaccgcccgcaccggagccatggtgacggtcgagggcagcg 
               
               
                   
                 ggaaaacgggtcgataccctctcgaacagagtatgaatcgtctgagtcat 
               
               
                   
                 cttttggcaacggcagtggcagtagaaaataccagcacagatatgatgga 
               
               
                   
                 agttcacgtgacgcgccagcagcattatttcactgcgcgactgtcagata 
               
               
                   
                 tttatcagtatcccggattggatattgccttacagccggatgaccgtatc 
               
               
                   
                 actctgcgtcaagtgaccgagtatgttaatgtgctcggcgcagcaggtgt 
               
               
                   
                 tcaggggaaacatgctctggttcagcgtcattccagcgtcgtggatgctt 
               
               
                   
                 tggcgctcgcgaaaggattaaatgacaatcttgccgatccgcaagcgatt 
               
               
                   
                 tttctgtacaagcataacgaagcggaacaggcgaaacagcagatgcgtaa 
               
               
                   
                 gctgaatatctatcatgtcgatatgagccaacctaactcggtgtttttag 
               
               
                   
                 cgcaggccatacgagtggacaacggcgatgttatctatatctcgaacgcc 
               
               
                   
                 tctttgactgatttcgctaaggtgaaagccgcattcgatagctttttgac 
               
               
                   
                 ccgcggcactaattctttctaa 
               
               
                   
               
               
                 SEQ ID NO: 7 (Ty21a- 
                 cccgttcctcattgatttgattgctaacgtcatgagcattactctatcat 
               
               
                 M-N Construct) 
                 tccagaatgcctcaatgaacaagttgtttgagaaagagtgtcgcaatgtt 
               
               
                   
                 gcaaccagagcccttaaatatgtacgccagaagaaaactgaagggcgtct 
               
               
                   
                 ggatgaagcattgtctgtattgattagcctgaaacgaattgagcctgatg 
               
               
                   
                 tttctcgtctgatgcgtgaatataagcaaattatcagattatttaatgag 
               
               
                   
                 tcacggaaggatggcggtagcactatcacgtcttatgaacatctagacta 
               
               
                   
                 tgcgaaaaaattactcgtttttgatagcgaaaatgcctatgccttgaaat 
               
               
                   
                 atgccgcattaaatgcaatgcatttacgcgactacacgcaggctttgcag 
               
               
                   
                 tattggcagcgactggagaaagtgaatggaccaacggagccggtgacaag 
               
               
                   
                 gcagatctcgacctgcataaccgcattacaaaaaaatacatcagggaagt 
               
               
                   
                 cgtaacccggggtgtaggctggagctgcttcgaagttcctatactttcta 
               
               
                   
                 gagaataggaacttcggaataggaacttcaagatcccctcacgctgccgc 
               
               
                   
                 aagcactcagggcgcaagggctgctaaaggaagcggaacacgtagaaagc 
               
               
                   
                 cagtccgcagaaacggtgctgaccccggatgaatgtcagctactgggcta 
               
               
                   
                 tctggacaagggaaaacgcaagcgcaaagagaaagcaggtagcttgcagt 
               
               
                   
                 gggcttacatggcgatagctagactgggcggttttatggacagcaagcga 
               
               
                   
                 accggaattgccagctggggcgccctctggtaaggttgggaagccctgca 
               
               
                   
                 aagtaaactggatggctttcttgccgccaaggatctgatggcgcagggga 
               
               
                   
                 tcaagatctgatcaagagacaggatgaggatcgtttcgcatgattgaaca 
               
               
                   
                 agatggattgcacgcaggttctccggccgcttgggtggagaggctattcg 
               
               
                   
                 gctatgactgggcacaacagacaatcggctgctctgatgccgccgtgttc 
               
               
                   
                 cggctgtcagcgcaggggcgcccggttctttttgtcaagaccgacctgtc 
               
               
                   
                 cggtgccctgaatgaactgcaggacgaggcagcgcggctatcgtggctgg 
               
               
                   
                 ccacgacgggcgttccttgcgcagctgtgctcgacgttgtcactgaagcg 
               
               
                   
                 ggaagggactggctgctattgggcgaagtgccggggcaggatctcctgtc 
               
               
                   
                 atctcaccttgctcctgccgagaaagtatccatcatggctgatgcaatgc 
               
               
                   
                 ggcggctgcatacgcttgatccggctacctgcccattcgaccaccaagcg 
               
               
                   
                 aaacatcgcatcgagcgagcacgtactcggatggaagccggtcttgtcga 
               
               
                   
                 tcaggatgatctggacgaagagcatcaggggctcgcgccagccgaactgt 
               
               
                   
                 tcgccaggctcaaggcgcgcatgcccgacggcgaggatctcgtcgtgacc 
               
               
                   
                 catggcgatgcctgcttgccgaatatcatggtggaaaatggccgcttttc 
               
               
                   
                 tggattcatcgactgtggccggctgggtgtggcggaccgctatcaggaca 
               
               
                   
                 tagcgttggctacccgtgatattgctgaagagcttggcggcgaatgggct 
               
               
                   
                 gaccgcttcctcgtgctttacggtatcgccgctcccgattcgcagcgcat 
               
               
                   
                 cgccttctatcgccttcttgacgagttcttctgagcgggactctggggtt 
               
               
                   
                 cgaaatgaccgaccaagcgacgcccaacctgccatcacgagatttcgatt 
               
               
                   
                 ccaccgccgccttctatgaaaggttgggcttcggaatcgttttccgggac 
               
               
                   
                 gccggctggatgatcctccagcgcggggatctcatgctggagttcttcgc 
               
               
                   
                 ccaccccagcttcaaaagcgctctgaagttcctatactttctagagaata 
               
               
                   
                 ggaacttcggaataggaactaaggaggatattcatatatgcattaatgtc 
               
               
                   
                 taacaattcgttcaagccgacgccgcttcgcggcgcggcttaactcaagc 
               
               
                   
                 gttagatgcactaagcacataattgctcacagccaaactatcaggtcaag 
               
               
                   
                 tctgcttttattatttttaagcgtgcataataagccctacacaaattggg 
               
               
                   
                 agatatatcatgaaaggctggctttttcttgttatcgcaatagttggcga 
               
               
                   
                 agtaatcgcaacatccgcattaaaatctagcgagggctttactgtcgacc 
               
               
                   
                 aaaaaaatattgacaacataaaaaactttgtgttatacttgtaacgGact 
               
               
                   
                 ctagagggtattaataatgaaaggagaggacatGAGCTCCCTAGGATGAG 
               
               
                   
                 CGATAACGGCCCGCAGAACCAGCGCAACGCGCCGCGCATTACCTTTGGCG 
               
               
                   
                 GCCCGAGCGATAGCACCGGCAGCAACCAGAACGGCGAACGCAGCGGCGCG 
               
               
                   
                 CGCAGCAAACAGCGCCGCCCGCAGGGCCTGCCGAACAACACCGCGAGCTG 
               
               
                   
                 GTTTACCGCGCTGACCCAGCATGGCAAAGAAGATCTGAAATTTCCGCGCG 
               
               
                   
                 GCCAGGGCGTGCCGATTAACACCAACAGCAGCCCGGATGATCAGATTGGC 
               
               
                   
                 TATTATCGCCGCGCGACCCGCCGCATTCGCGGCGGCGATGGCAAAATGAA 
               
               
                   
                 AGATCTGAGCCCGCGCTGGTATTTTTATTATCTGGGCACCGGCCCGGAAG 
               
               
                   
                 CGGGCCTGCCGTATGGCGCGAACAAAGATGGCATTATTTGGGTGGCGACC 
               
               
                   
                 GAAGGCGCGCTGAACACCCCGAAAGATCATATTGGCACCCGCAACCCGGC 
               
               
                   
                 GAACAACGCGGCGATTGTGCTGCAGCTGCCGCAGGGCACCACCCTGCCGA 
               
               
                   
                 AAGGCTTTTATGCGGAAGGCAGCCGCGGCGGCAGCCAGGCGAGCAGCCGC 
               
               
                   
                 AGCAGCAGCCGCAGCCGCAACAGCAGCCGCAACAGCACCCCGGGCAGCAG 
               
               
                   
                 CCGCGGCACCAGCCCGGCGCGCATGGCGGGCAACGGCGGCGATGCGGCGC 
               
               
                   
                 TGGCGCTGCTGCTGCTGGATCGCCTGAACCAGCTGGAAAGCAAAATGAGC 
               
               
                   
                 GGCAAAGGCCAGCAGCAGCAGGGCCAGACCGTGACCAAAAAAAGCGCGGC 
               
               
                   
                 GGAAGCGAGCAAAAAACCGCGCCAGAAACGCACCGCGACCAAAGCGTATA 
               
               
                   
                 ACGTGACCCAGGCGTTTGGCCGCCGCGGCCCGGAACAGACCCAGGGCAAC 
               
               
                   
                 TTTGGCGATCAGGAACTGATTCGCCAGGGCACCGATTATAAACATTGGCC 
               
               
                   
                 GCAGATTGCGCAGTTTGCGCCGAGCGCGAGCGCGTTTTTTGGCATGAGCC 
               
               
                   
                 GCATTGGCATGGAAGTGACCCCGAGCGGCACCTGGCTGACCTATACCGGC 
               
               
                   
                 GCGATTAAACTGGATGATAAAGATCCGAACTTTAAAGATCAGGTGATTCT 
               
               
                   
                 GCTGAACAAACATATTGATGCGTATAAAACCTTTCCGCCGACCGAACCGA 
               
               
                   
                 AAAAAGATAAAAAAAAAAAAGCGGATGAAACCCAGGCGCTGCCGCAGCGC 
               
               
                   
                 CAGAAAAAACAGCAGACCGTGACCCTGCTGCCGGCGGCGGATCTGGATGA 
               
               
                   
                 TTTTAGCAAACAGCTGCAGCAGAGCATGAGCAGCGCGGATAGCACCCAGG 
               
               
                   
                 CGCCTAGGCTTAAGATGGCGGATAGCAACGGCACCATTACCGTGGAAGAA 
               
               
                   
                 CTGAAAAAACTGCTGGAACAGTGGAACCTGGTGATTGGCTTTCTGTTTCT 
               
               
                   
                 GACCTGGATTTGCCTGCTGCAGTTTGCGTATGCGAACCGCAACCGCTTTC 
               
               
                   
                 TGTATATTATTAAACTGATTTTTCTGTGGCTGCTGTGGCCGGTGACCCTG 
               
               
                   
                 GCGTGCTTTGTGCTGGCGGCGGTGTATCGCATTAACTGGATTACCGGCGG 
               
               
                   
                 CATTGCGATTGCGATGGCGTGCCTGGTGGGCCTGATGTGGCTGAGCTATT 
               
               
                   
                 TTATTGCGAGCTTTCGCCTGTTTGCGCGCACCCGCAGCATGTGGAGCTTT 
               
               
                   
                 AACCCGGAAACCAACATTCTGCTGAACGTGCCGCTGCATGGCACCATTCT 
               
               
                   
                 GACCCGCCCGCTGCTGGAAAGCGAACTGGTGATTGGCGCGGTGATTCTGC 
               
               
                   
                 GCGGCCATCTGCGCATTGCGGGCCATCATCTGGGCCGCTGCGATATTAAA 
               
               
                   
                 GATCTGCCGAAAGAAATTACCGTGGCGACCAGCCGCACCCTGAGCTATTA 
               
               
                   
                 TAAACTGGGCGCGAGCCAGCGCGTGGCGGGCGATAGCGGCTTTGCGGCGT 
               
               
                   
                 ATAGCCGCTATCGCATTGGCAACTATAAACTGAACACCGATCATAGCAGC 
               
               
                   
                 AGCAGCGATAACATTGCGCTGCTGGTGCAGCTTAAGGCATTAGCCTATGG 
               
               
                   
                 AAGTCAGGGTAATCTTAATCCATTAATTAATGAAATCAGCAAAATCATTT 
               
               
                   
                 CAGCTGCAGGTAATTTTGATGTTAAAGAGGAAAGAGCGGCCGCGTCTTTA 
               
               
                   
                 TTGCAGTTGTCCGGTAATGCCAGTGATTTTTCATATGGACGGAACTCAAT 
               
               
                   
                 AACTTTGACAGCATCAGCATAAGAGCTCtttattaatttaaataatagca 
               
               
                   
                 atcttactgggctgtgccacataagattgctatttttttggagtcataat 
               
               
                   
                 ggattcttgtcataaaattgattatgggttatacgccctggagattttag 
               
               
                   
                 cccaataccataacgtctctgttaacccggaagaaattaaacatagattt 
               
               
                   
                 gacacagacgggactggtctgggattaacgtcatggttgcttgctgcgaa 
               
               
                   
                 atctttagaactaaaggtaaaacaggtaaaaaaaacaattgaccgattaa 
               
               
                   
                 actttatttctctgcccgcattagtctggagagaggatggatgtcatttt 
               
               
                   
                 attctgactaaagtcagtaaagaagcaaacagatatcttatttttgatct 
               
               
                   
                 ggagcagcgaaatccccgtgttctcgaacagtctgagtttgaggcgttat 
               
               
                   
                 atcaggggcatattattcttattgcttcccgttcttctgttaccgggaaa 
               
               
                   
                 ctggcaaaatttgactttacctggtttattcctgccattataaaatacag 
               
               
                   
                 gaaaatatttattgaaacccttgttgtatctgtttttttacaattatttg 
               
               
                   
                 cattaataaccccccttttttttcaggtggttatggacaaagtattagta 
               
               
                   
                 cacagggggttttcaacccttaatgttattactgtcgcattatctgttgt 
               
               
                   
                 ggtggtgtttgagattatactcagcggtttaagaacttacatttttgcac 
               
               
                   
                 atagtacaagtcggattgatgttgagttgggtgccaaactcttccggcat 
               
               
                   
                 ttactggcgctaccgatctcttattttgagagtcgtcgtgttggtgatac 
               
               
                   
                 tgttgccagggtaagagaattagaccagatccgtaatttcctgacaggac 
               
               
                   
                 aggcattaacatctgttctggacttattattttcattcatattttttgcg 
               
               
                   
                 gtaatgtggtattacagcccaaagcttactctggtgatcttattttcgct 
               
               
                   
                 gccctgttatgctgcatggtctgtttttattagccccattttgcgacgtc 
               
               
                   
                 gccttgatgataagttttcacggaatgcggataatcaatctttcctggtg 
               
               
                   
                 gaatcagtcacggcgattaacactataaaagctatggcagtctcacctca 
               
               
                   
                 gatgacgaacatatgggacaaacaattggcaggatatgttgctgcaggct 
               
               
                   
                 ttaaagtgacagtattagccaccattggtcaacaaggaatacagttaata 
               
               
                   
                 caaaagactgttatgatcatcaacctgtggttgggtgcacacctggttat 
               
               
                   
                 ttccggggatttaagtattggtcagttaattgcttttaatatgcttgcag 
               
               
                   
                 gtcagattgttgcaccggttattcgccttgcacaaatctggcaggatttc 
               
               
                   
                 cagcaggttggtatatcagttacccgccttggtgatgtgcttaactctcc 
               
               
                   
                 aactgaaagttatcatgggaaactggcattaccggaaattaatggtgata 
               
               
                   
                 tcacttttcgtaatatccggtttcgctataagcctgactctccggttatt 
               
               
                   
                 ttagataatatcaatctcagtattaagcagggggaggttattggtattgt 
               
               
                   
                 cggacgttctggttcaggaaaaagcacattaactaaattaattcaacgtt 
               
               
                   
                 tttatattcctgaaaatggccaggtcttaattgatggacatgatcttgcg 
               
               
                   
                 ttggccgatcctaactggttacgtcgtcaggtgggggttgtgttgcagga 
               
               
                   
                 caatgtgctgcttaatcgcagtattattgataatatttcactggctaatc 
               
               
                   
                 ctggcatgtccgtcgaaaaagttatttatgcagcgaaattagcaggtgct 
               
               
                   
                 catgattttatttctgaattgcgtgaggggtataacaccattgtcgggga 
               
               
                   
                 acagggggcaggattatccggaggtcaacgtcaacgcatcgcaattgcaa 
               
               
                   
                 gggcgctggtgaacaaccctaaaatactcatctttgatgaagcaaccagt 
               
               
                   
                 gctctggattatgagtcggagcatgtcatcatgcgcaatatgcacaaaat 
               
               
                   
                 atgtaagggcagaacggttataatcattgctcatcgtctgtctacagtaa 
               
               
                   
                 aaaatgcagaccgcattattgtcatggaaaaagggaaaattgttgaacag 
               
               
                   
                 ggtaaacataaggagctgctttctgaaccggaaagtttatacagttactt 
               
               
                   
                 atatcagttacagtcagactaacagaaagaacagaagaatatgaaaacat 
               
               
                   
                 ggttaatggggttcagcgagttcctgttgcgctataaacttgtctggagt 
               
               
                   
                 gaaacatggaaaatccggaagcagttagatactccggtacgtgaaaagga 
               
               
                   
                 cgaaaatgaattcttacccgctcatctggaattaattgaaacgccggtat 
               
               
                   
                 ccagacggccgcgtctggttgcttattttattatggggtttctggttatt 
               
               
                   
                 gctttcattttatctgttttaggccaggtggaaattgttgccactgcaaa 
               
               
                   
                 tgggaaattaacactcagtgggcgtagcaaagaaattaaacctattgaaa 
               
               
                   
                 actcgatagttaaagaaattatcgtaaaagaaggagagtcagtccggaaa 
               
               
                   
                 ggggatgtgttattaaagcttacagcgctgggagctgaagctgatacgtt 
               
               
                   
                 aaaaacgcagtcatcactgttacaggccaggctggaacaaattcggtatc 
               
               
                   
                 aaattctgagccggtcaattgaattaaataaacttcctgaactgaaactt 
               
               
                   
                 cctgatgagccttattttcagaatgtatctgaagaggaagtactgcgttt 
               
               
                   
                 aacttctttgataaaagaacagttttccacatggcaaaatcagaagtatc 
               
               
                   
                 aaaaagaactgaatctggataagaaaagagcagagcgattaacaatactt 
               
               
                   
                 gcccgtataaaccgttatgaaaatgtatcgagggttgaaaaaagccgtct 
               
               
                   
                 ggatgatttcaggagcctgttgcataaacaggcaattgcaaaacatgctg 
               
               
                   
                 tacttgagcaggagaataaatatgttgaggcagcaaatgaattacgggtt 
               
               
                   
                 tataaatcgcaactggagcaaattgagagtgagatattgtctgcaaaaga 
               
               
                   
                 agaatatcagcttgtcacgcagctttttaaaaatgaaattttagacaagc 
               
               
                   
                 taagacaaacaacagacagcattgagttattaactctggagttagagaaa 
               
               
                   
                 aatgaagagcgtcaacaggcttcagtaatcagggcccctgtttcgggaaa 
               
               
                   
                 agttcagcaactgaaggttcatactgaaggtggggttgttacaacagcgg 
               
               
                   
                 aaacactgatggtcatcgttccggaagatgacacgctggaggttactgct 
               
               
                   
                 ctggtacaaaataaagatattggttttattaacgtcgggcagaatgccat 
               
               
                   
                 cattaaagtggaggcttttccttacacccgatatggttatctggtgggta 
               
               
                   
                 aggtaaaaaatataaatttagatgcaatagaagaccagaaactgggactc 
               
               
                   
                 gtttttaatgtcattgtttctgttgaagagaatgatttgtcaaccgggaa 
               
               
                   
                 taagcacattccattaagctcgggtatggctgtcactgcagaaataaaga 
               
               
                   
                 ctggaatgcgaagcgtaatcagttatcttcttagtcccctggaagaatct 
               
               
                   
                 gtaacagaaagtttacatgagcgttaagtctcagagcagcggtatccggc 
               
               
                   
                 tcatatcttctcctgtcagatccaagggcgaattccagcacactggcggc 
               
               
                   
                 cgttactagtggatccaccggtctcgaggggcgcgccggtaccgaattct 
               
               
                   
                 aattaatgggcatcatttttcagctatttcatttataaaataagttatga 
               
               
                   
                 aaaaaatcatcatattactaacgacatttttcctgctttcgggatgcact 
               
               
                   
                 attcccagggcggtatttaaatccagccttattaatcaggacgatcctcg 
               
               
                   
                 ttataatctggtcgaagtcacgccgacattaaaactaagcgctcccgata 
               
               
                   
                 ctgtgccgaaaactattgtcgatccggtttttgccgcaaataactggcac 
               
               
                   
                 tggacatctttggctaaaggcgatgtgctgcatatcactattttatcctc 
               
               
                   
                 gggcggggctggatatttatccaataacgcgagcggcgaccgtgcggatt 
               
               
                   
                 ttgaaaatattcttgtgactgacagtaataccgttcaggtgccttatgcc 
               
               
                   
                 gggacaatcccggtttctggattggatgtgacgcaactggctgatgagat 
               
               
                   
                 caaaaagcgactttcgcgcgttgtcctgaatcctcaggtgattgtgacac 
               
               
                   
                 ttaccgcccgcaccggagccatggtgacggtcgagggcagcgggaaaacg 
               
               
                   
                 ggtcgataccctctcgaacagagtatgaatcgtctgagtcatcttttggc 
               
               
                   
                 aacggcagtggcagtagaaaataccagcacagatatgatggaagttcacg 
               
               
                   
                 tgacgcgccagcagcattatttcactgcgcgactgtcagatatttatcag 
               
               
                   
                 tatcccggattggatattgccttacagccggatgaccgtatcactctgcg 
               
               
                   
                 tcaagtgaccgagtatgttaatgtgctcggcgcagcaggtgttcagggga 
               
               
                   
                 aacatgctctggttcagcgtcattccagcgtcgtggatgctttggcgctc 
               
               
                   
                 gcgaaaggattaaatgacaatcttgccgatccgcaagcgatttttctgta 
               
               
                   
                 caagcataacgaagcggaacaggcgaaacagcagatgcgtaagctgaata 
               
               
                   
                 tctatcatgtcgatatgagccaacctaactcggtgtttttagcgcaggcc 
               
               
                   
                 atacgagtggacaacggcgatgttatctatatctcgaacgcctctttgac 
               
               
                   
                 tgatttcgctaaggtgaaagccgcattcgatagctttttgac 
               
               
                   
               
               
                 SEQ ID NO: 8 (Ty21a- 
                 cccgttcctcattgatttgattgctaacgtcatgagcattactctatcat 
               
               
                 eS-AR Construct) 
                 tccagaatgcctcaatgaacaagttgtttgagaaagagtgtcgcaatgtt 
               
               
                   
                 gcaaccagagcccttaaatatgtacgccagaagaaaactgaagggcgtct 
               
               
                   
                 ggatgaagcattgtctgtattgattagcctgaaacgaattgagcctgatg 
               
               
                   
                 tttctcgtctgatgcgtgaatataagcaaattatcagattatttaatgag 
               
               
                   
                 tcacggaaggatggcggtagcactatcacgtcttatgaacatctagacta 
               
               
                   
                 tgcgaaaaaattactcgtttttgatagcgaaaatgcctatgccttgaaat 
               
               
                   
                 atgccgcattaaatgcaatgcatttacgcgactacacgcaggctttgcag 
               
               
                   
                 tattggcagcgactggagaaagtgaatggaccaacggagccggtgacaag 
               
               
                   
                 gcagatctcgacctgcataaccgcattacaaaaaaatacatcagggaagt 
               
               
                   
                 cgtaacccggggtgtaggctggagctgcttcgaagttcctatactttcta 
               
               
                   
                 gagaataggaacttcggaataggaacttcaagatcccctcacgctgccgc 
               
               
                   
                 aagcactcagggcgcaagggctgctaaaggaagcggaacacgtagaaagc 
               
               
                   
                 cagtccgcagaaacggtgctgaccccggatgaatgtcagctactgggcta 
               
               
                   
                 tctggacaagggaaaacgcaagcgcaaagagaaagcaggtagcttgcagt 
               
               
                   
                 gggcttacatggcgatagctagactgggcggttttatggacagcaagcga 
               
               
                   
                 accggaattgccagctggggcgccctctggtaaggttgggaagccctgca 
               
               
                   
                 aagtaaactggatggctttcttgccgccaaggatctgatggcgcagggga 
               
               
                   
                 tcaagatctgatcaagagacaggatgaggatcgtttcgcatgattgaaca 
               
               
                   
                 agatggattgcacgcaggttctccggccgcttgggtggagaggctattcg 
               
               
                   
                 gctatgactgggcacaacagacaatcggctgctctgatgccgccgtgttc 
               
               
                   
                 cggctgtcagcgcaggggcgcccggttctttttgtcaagaccgacctgtc 
               
               
                   
                 cggtgccctgaatgaactgcaggacgaggcagcgcggctatcgtggctgg 
               
               
                   
                 ccacgacgggcgttccttgcgcagctgtgctcgacgttgtcactgaagcg 
               
               
                   
                 ggaagggactggctgctattgggcgaagtgccggggcaggatctcctgtc 
               
               
                   
                 atctcaccttgctcctgccgagaaagtatccatcatggctgatgcaatgc 
               
               
                   
                 ggcggctgcatacgcttgatccggctacctgcccattcgaccaccaagcg 
               
               
                   
                 aaacatcgcatcgagcgagcacgtactcggatggaagccggtcttgtcga 
               
               
                   
                 tcaggatgatctggacgaagagcatcaggggctcgcgccagccgaactgt 
               
               
                   
                 tcgccaggctcaaggcgcgcatgcccgacggcgaggatctcgtcgtgacc 
               
               
                   
                 catggcgatgcctgcttgccgaatatcatggtggaaaatggccgcttttc 
               
               
                   
                 tggattcatcgactgtggccggctgggtgtggcggaccgctatcaggaca 
               
               
                   
                 tagcgttggctacccgtgatattgctgaagagcttggcggcgaatgggct 
               
               
                   
                 gaccgcttcctcgtgctttacggtatcgccgctcccgattcgcagcgcat 
               
               
                   
                 cgccttctatcgccttcttgacgagttcttctgagcgggactctggggtt 
               
               
                   
                 cgaaatgaccgaccaagcgacgcccaacctgccatcacgagatttcgatt 
               
               
                   
                 ccaccgccgccttctatgaaaggttgggcttcggaatcgttttccgggac 
               
               
                   
                 gccggctggatgatcctccagcgcggggatctcatgctggagttcttcgc 
               
               
                   
                 ccaccccagcttcaaaagcgctctgaagttcctatactttctagagaata 
               
               
                   
                 ggaacttcggaataggaactaaggaggatattcatatatgcattaatgtc 
               
               
                   
                 taacaattcgttcaagccgacgccgcttcgcggcgcggcttaactcaagc 
               
               
                   
                 gttagatgcactaagcacataattgctcacagccaaactatcaggtcaag 
               
               
                   
                 tctgcttttattatttttaagcgtgcataataagccctacacaaattggg 
               
               
                   
                 agatatatcatgaaaggctggctttttcttgttatcgcaatagttgCGAT 
               
               
                   
                 CGTTATGACAACTTGACGGCTACATCATTCACTTTTTCTTCACAACCGGC 
               
               
                   
                 ACGGAACTCGCTCGGGCTGGCCCCGGTGCATTTTTTAAATACCCGCGAGA 
               
               
                   
                 AGTAGAGTTGATCGTCAAAACCAACATTGCGACCGACGGTGGCGATAGGC 
               
               
                   
                 ATCCGGGTGGTGCTCAAAAGCAGCTTCGCCTGGCTGATACGTTGGTCCTC 
               
               
                   
                 GCGCCAGCTTAAGACGCTAATCCCTAACTGCTGGCGGAAAAGATGTGACA 
               
               
                   
                 GACGCGACGGCGACAAGCAAACATGCTGTGCGACGCTGGCGATATCAAAA 
               
               
                   
                 TTGCTGTCTGCCAGGTGATCGCTGATGTACTGACAAGCCTCGCGTACCCG 
               
               
                   
                 ATTATCCATCGGTGGATGGAGCGACTCGTTAATCGCTTCCATGCGCCGCA 
               
               
                   
                 GTAACAATTGCTCAAGCAGATTTATCGCCAGCAGCTCCGAATAGCGCCCT 
               
               
                   
                 TCCCCTTGCCCGGCGTTAATGATTTGCCCAAACAGGTCGCTGAAATGCGG 
               
               
                   
                 CTGGTGCGCTTCATCCGGGCGAAAGAACCCCGTATTGGCAAATATTGACG 
               
               
                   
                 GCCAGTTAAGCCATTCATGCCAGTAGGCGCGCGGACGAAAGTAAACCCAC 
               
               
                   
                 TGGTGATACCATTCGCGAGCCTCCGGATGACGACCGTAGTGATGAATCTC 
               
               
                   
                 TCCTGGCGGGAACAGCAAAATAACACCCGGTCGGCAAACAAATTCTCGTC 
               
               
                   
                 CCTGATTTTTCACCACCCCCTGACCGCGAATGGTGAGATTGAGAATATAA 
               
               
                   
                 CCTTTCATTCCCAGCGGTCGGTCGATAAAAAAATCGAGATAACCGTTGGC 
               
               
                   
                 CTCAATCGGCGTTAAACCCGCCACCAGATGGGCATTAAACGAGTATCCCG 
               
               
                   
                 GCAGCAGGGGATCATTTTGCGCTTCAGCCATACTTTTCATACTCCCGCCA 
               
               
                   
                 TTCAGAGAAGAAACCAATTGTCCATATTGCATCAGACATTGCCGTCACTG 
               
               
                   
                 CGTCTTTTACTGGCTCTTCTCGCTAACCAAACCGGTAACCCCGCTTATTA 
               
               
                   
                 AAAGCATTCTGTAACAAAGCGGGACCAAAGCCATGACAAAAACGCGTAAC 
               
               
                   
                 AAAAGTGTCTATAATCACGGCAGAAAAGTCCACATTGATTATTTGCACGG 
               
               
                   
                 CGTCACACTTTGCTATGCCATAGCATTTTTATCCATAAGATTAGCGGATC 
               
               
                   
                 CTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACCCGTTT 
               
               
                   
                 TTTTGGGAATTCGAGCTCGGAGTTAACAAAAGATGTTAGATGCAAACAAA 
               
               
                   
                 TTACAGCAGGCAGTGGATCAGGCTTACACCCAATTTCACTCACTTAACGG 
               
               
                   
                 CGGACAAAATGCCGATTACATTCCCTTTCTGGCGAATGTACCAGGTCAAC 
               
               
                   
                 TGGCGGCAGTGGCTATCGTGACNTGCGATGGCAACGTCTATAGTGCGGGT 
               
               
                   
                 GACAGTGATTACCGCTTTGCACTGGAATCCATCTCTAAAGTCTGTACGTT 
               
               
                   
                 AGCCCTTGCGTTAGAAGATGTCGGCCCGCAGGCGGTACAGGACAAAATTG 
               
               
                   
                 GCGCTGACCCGACCGGATTGCCCTTTAACTCAGTTATCGCCTTAGAGTTG 
               
               
                   
                 CATGGCGGCAAACCGCTGTCGCCACTGGTAAATGCTGGCGCTATTGCCAC 
               
               
                   
                 CACCAGCCTGATTAACGCTGAAAATGTTGAACAACGCTGGCAGCGAATTT 
               
               
                   
                 TACATATCCAACAGCAACTGGCTGGTGAGCAGGTAGCGCTCTCTGACGAA 
               
               
                   
                 GTCAACCAGTCGGAACAAACAACCAACTTCCATAACCGGGCCATTGCCTG 
               
               
                   
                 GCTGCTGTACTCCGCCGGATATCTCTATTGTGATGCAATGGAAGCCTGTG 
               
               
                   
                 ACGTGTACACCCGTCAGTGCTCTACGCTCATCAATACTGTTGAACTGGCA 
               
               
                   
                 ACGCTTGGCGCGACGCTGGCGGCGGGTGGTGTGAATCCGTTGACGCATAA 
               
               
                   
                 ACGCGTTCTTCAGGCCGACAACGTGCCGTACATTCTGGCCGAAATGATGA 
               
               
                   
                 TGGAAGGGCTGTATGGTCGCTCCGGTGACTGGGCGTATCGCGTTGGTTTA 
               
               
                   
                 CCGGGCAAAAGCGGTGTAGGTGGCGGTATTCTGGCGGTCGTCCCTGGCGT 
               
               
                   
                 GATGGGAATTGCCGCGTTCTCACCACCGCTGGACGAAGAAGGCAACAGTG 
               
               
                   
                 TTCGCGGTCAAAAAATGGTGGCATCGGTCGCTAAGCAACTCGGCTATAAC 
               
               
                   
                 GTGTTTAAGGGCTGACCATGGAGTGTTTTAATGCGATCCAATCATTTTAA 
               
               
                   
                 GGAGTTTAAAATGGATAAGAAGCAAGTAACGGATTTAAGGTCGGAACTAC 
               
               
                   
                 TCGATTCACGTTTTGGTGCGAAGTCTATTTCCACTATCGCAGAATCAAAA 
               
               
                   
                 CGTTTTCCGCTGCACGAAATGCGCGACGATGTCGCATTTCAGATTATCAA 
               
               
                   
                 TGATGAATTATATCTTGATGGCAACGCTCGTCAGAACCTGGCCACTTTCT 
               
               
                   
                 GCCAGACCTGGGACGACGAAAACGTCCACAAGTTGATGGATTTATCCATT 
               
               
                   
                 AACAAAAACTGGATCGACAAAGAAGAATATCCGCAATCCGCAGCCATCGA 
               
               
                   
                 CCTGCGTTGCGTAAACATGGTTGCCGATCTGTGGCATGCGCCTGCGCCGA 
               
               
                   
                 AAAATGGTCAGGCCGTTGGCACCAACACCATTGGTTCTTCCGAGGCCTGT 
               
               
                   
                 ATGCTCGGCGGGATGGCGATGAAATGGCGTTGGCGCAAGCGTATGGAAGC 
               
               
                   
                 TGCAGGCAAACCAACGGATAAACCAAACCTGGTGTGCGGTCCGGTGCAAA 
               
               
                   
                 TCTGCTGGCATAAATTCGCCCGCTACTGGGATGTGGAGCTGCGTGAGATC 
               
               
                   
                 CCTATGCGCCCCGGTCAGTTGTTTATGGACCCGAAACGCATGATTGAAGC 
               
               
                   
                 CTGCGACGAAAATACCATCGGCGTGGTGCCGACTTTCGGCGTGACCTACA 
               
               
                   
                 CCGGTAACTATGAGTTCCCGCAACCGCTGCACGATGCACTGGATAAATTC 
               
               
                   
                 CAGGCCGACACCGGTATCGACATCGACATGCACATCGACGCCGCCAGCGG 
               
               
                   
                 TGGCTTTCTGGCACCGTTCGTCGCCCCGGATATCGTCTGGGACTTCCGCC 
               
               
                   
                 TGCCGCGTGTGAAATCGATCAGTGCTTCAGGCCATAAATTCGGTCTGGCT 
               
               
                   
                 CCGCTGGGCTGCGGCTGGGTTATCTGGCGTGACGAAGAAGCGCTGCCGCA 
               
               
                   
                 GGAACTGGTGTTCAACGTTGACTACCTCGGTGGTCAGATTGGTACTTTTG 
               
               
                   
                 CCATCAACTTCTCCCGCCCGGCGGGTCAGGTGATTGCACAGTACTATGAA 
               
               
                   
                 TTCCTGCGCCTTGGTCGTGAAGGCTATACCAAAGTACAGAATGCTTCCTA 
               
               
                   
                 CCAGGTCGCTGCCTATCTGGCGGATGAGATCGCCAAACTGGGGCCGTATG 
               
               
                   
                 AGTTCATCTGTACCGGTCGCCCGGACGAAGGCATCCCGGCGGTTTGCTTC 
               
               
                   
                 AAACTGAAAGATGGTGAAGATCCGGGATACACCCTGTACGACCTCTCTGA 
               
               
                   
                 ACGTCTGCGTCTGCGCGGCTGGCAGGTTCCGGCCTTCACTCTCGGCGGTG 
               
               
                   
                 AAGCCACCGACATCGTGGTGATGCGCATTATGTGTCGTCGCGGCTTCGAA 
               
               
                   
                 ATGGACTTTGCTGAACTGTTGCTGGAAGACTACAAAGCCTCCCTGAAATA 
               
               
                   
                 TCTCAGCGATCACCCGAAACTGCAGGGTATTGCCCAGCAGAACAGCTTTA 
               
               
                   
                 AACATACCTGATAACCGTTTTGGAGTGATAATATGGCTACATCAGTACAG 
               
               
                   
                 ACAGGTAAAGCTAAGCAGCTCACATTACTCGGATTCTTTGCCATAACGGC 
               
               
                   
                 ATCGATGGTAATGGCTGTTTATGAATACCCTACCTTCGCAACATCGGGCT 
               
               
                   
                 TTTCATTAGTCTTCTTCCTGCTATTAGGCGGGATTTTATGGTTTATTCCC 
               
               
                   
                 GTGGGACTTTGTGCTGCGGAAATGGCCACCGTCGACGGCTGGGAAGAAGG 
               
               
                   
                 TGGTGTCTTCGCCTGGGTATCAAATACTCTGGGTCCGAGATGGGGATTTG 
               
               
                   
                 CAGCGATCTCATTTGGCTATCTGCAAATCGCCATTGGTTTTATTCCGATG 
               
               
                   
                 CTCTATTTCGTGTTAGGGGCACTCTCCTACATCCTGAAATGGCCAGCGCT 
               
               
                   
                 GAATGAAGACCCCATTACCAAAACTATTGCAGCACTCATCATTCTTTGGG 
               
               
                   
                 CGCTGGCATTAACGCAGTTTGGTGGCACGAAATACACGGCGCGAATTGCT 
               
               
                   
                 AAAGTTGGCTTCTTCGCCGGTATCCTGTTACCTGCATTTATTTTGATCGC 
               
               
                   
                 ATTAGCGGCTATTTACCTGCACTCCGGTGCCCCCGTTGCTATCGAAATGG 
               
               
                   
                 ATTCGAAGACCTTCTTCCCTGACTTCTCTAAAGTGGGCACACTGGTTGTA 
               
               
                   
                 TTTGTTGCCTTCATTTTGAGTTATATGGGCGTAGAAGCTTCCGCAACTCA 
               
               
                   
                 CGTCAATGAAATGAGCAACCCCGGGCGCGACTATCCACTGGCTATGTTAC 
               
               
                   
                 TGCTGATGGTGGCGGCAATCTGCTTAAGCTCTGTTGGCGGTTTGTCTATT 
               
               
                   
                 GCGATGGTCATTCCGGGTAATGAAATCAACCTCTCCGCAGGGGTAATGCA 
               
               
                   
                 AACCTTTACCGTTCTGATGTCCCATGTGGCACCGGAAATTGAGTGGACGG 
               
               
                   
                 TTCGCGTGATCTCCGCACTGCTGTTGCTGGGTGTTCTGGCGGAAATCGCC 
               
               
                   
                 TCCTGGATTGTTGGTCCTTCTCGCGGGATGTATGTCACAGCGCAGAAAAA 
               
               
                   
                 CCTGCTGCCAGCGGCATTCGCTAAAATGAACAAAAATGGCGTACCGGTAA 
               
               
                   
                 CGCTGGTCATTTCGCAGCTGGTGATTACTTCTATCGCGTTGATCATCCTC 
               
               
                   
                 ACCAATACCGGTGGCGGTAACAACATGTCCTTCCTGATCGCACTGGCGCT 
               
               
                   
                 GACGGTGGTGATTTATCTGTGTGCTTATTTCATGCTGTTTATTGGCTACA 
               
               
                   
                 TTGTGTTGGTTCTTAAACATCCTGACTTAAAACGCACATTTAATATCCCT 
               
               
                   
                 GGTGGTAAAGGGGTGAAACTGGTCGTGGCAATTGTCGGTCTGCTGACTTC 
               
               
                   
                 AATTATGGCGTTTATTGTTTCCTTCCTGCCGCCGGATAACATCCAGGGTG 
               
               
                   
                 ATTCTACCGATATGTATGTTGAATTACTGGTTGTTAGTTTCCTGGTGGTA 
               
               
                   
                 CTTGCCCTGCCCTTTATTCTCTATGCTGTTCATGATCGTAAAGGCAAAGC 
               
               
                   
                 GAATACCGGCGTCACTCTGGAGCCAATCAACAGTCAGAACGCACCAAAAG 
               
               
                   
                 GTCACTTCTTCCTGCACCCGCGTGCACGTTCACCACACTATATTGTGATG 
               
               
                   
                 AATGACAAGAAACACTAACGATCGgcgaagtaatcgcaacatccgcatta 
               
               
                   
                 aaatctagcgagggctttactgtcgaccaaaaaaatattgacaacataaa 
               
               
                   
                 aaactttgtgttatacttgtaacgGactctagagggtattaataatgaaa 
               
               
                   
                 ggagaggacatGAGCTCATGTTTGTGTTTCTGGTGCTGCTGCCGCTGGTG 
               
               
                   
                 AGCAGCCAGTGCGTGAACCTGACCACCCGCACCCAGCTGCCGCCGGCGTA 
               
               
                   
                 TACCAACAGCTTTACCCGCGGCGTGTATTATCCGGATAAAGTGTTTCGCA 
               
               
                   
                 GCAGCGTGCTGCATAGCACCCAGGATCTGTTTCTGCCGTTTTTTAGCAAC 
               
               
                   
                 GTGACCTGGTTTCATGCGATTCATGTGAGCGGCACCAACGGCACCAAACG 
               
               
                   
                 CTTTGATAACCCGGTGCTGCCGTTTAACGATGGCGTGTATTTTGCGAGCA 
               
               
                   
                 CCGAAAAAAGCAACATTATTCGCGGCTGGATTTTTGGCACCACCCTGGAT 
               
               
                   
                 AGCAAAACCCAGAGCCTGCTGATTGTGAACAACGCGACCAACGTGGTGAT 
               
               
                   
                 TAAAGTGTGCGAATTTCAGTTTTGCAACGATCCGTTTCTGGGCGTGTATT 
               
               
                   
                 ATCATAAAAACAACAAAAGCTGGATGGAAAGCGAATTTCGCGTGTATAGC 
               
               
                   
                 AGCGCGAACAACTGCACCTTTGAATATGTGAGCCAGCCGTTTCTGATGGA 
               
               
                   
                 TCTGGAAGGCAAACAGGGCAACTTTAAAAACCTGCGCGAATTTGTGTTTA 
               
               
                   
                 AAAACATTGATGGCTATTTTAAAATTTATAGCAAACATACCCCGATTAAC 
               
               
                   
                 CTGGTGCGCGATCTGCCGCAGGGCTTTAGCGCGCTGGAACCGCTGGTGGA 
               
               
                   
                 TCTGCCGATTGGCATTAACATTACCCGCTTTCAGACCCTGCTGGCGCTGC 
               
               
                   
                 ATCGCAGCTATCTGACCCCGGGCGATAGCAGCAGCGGCTGGACCGCGGGC 
               
               
                   
                 GCGGCGGCGTATTATGTGGGCTATCTGCAGCCGCGCACCTTTCTGCTGAA 
               
               
                   
                 ATATAACGAAAACGGCACCATTACCGATGCGGTGGATTGCGCGCTGGATC 
               
               
                   
                 CGCTGAGCGAAACCAAATGCACCCTGAAAAGCTTTACCGTGGAAAAAGGC 
               
               
                   
                 ATTTATCAGACCAGCAACTTTCGCGTGCAGCCGACCGAAAGCATTGTGCG 
               
               
                   
                 CTTTCCGAACATTACCAACCTGTGCCCGTTTGGCGAAGTGTTTAACGCGA 
               
               
                   
                 CCCGCTTTGCGAGCGTGTATGCGTGGAACCGCAAACGCATTAGCAACTGC 
               
               
                   
                 GTGGCGGATTATAGCGTGCTGTATAACAGCGCGAGCTTTAGCACCTTTAA 
               
               
                   
                 ATGCTATGGCGTGAGCCCGACCAAACTGAACGATCTGTGCTTTACCAACG 
               
               
                   
                 TGTATGCGGATAGCTTTGTGATTCGCGGCGATGAAGTGCGCCAGATTGCG 
               
               
                   
                 CCGGGCCAGACCGGCAAAATTGCGGATTATAACTATAAACTGCCGGATGA 
               
               
                   
                 TTTTACCGGCTGCGTGATTGCGTGGAACAGCAACAACCTGGATAGCAAAG 
               
               
                   
                 TGGGCGGCAACTATAACTATCTGTATCGCCTGTTTCGCAAAAGCAACCTG 
               
               
                   
                 AAACCGTTTGAACGCGATATTAGCACCGAAATTTATCAGGCGGGCAGCAC 
               
               
                   
                 CCCGTGCAACGGCGTGGAAGGCTTTAACTGCTATTTTCCGCTGCAGAGCT 
               
               
                   
                 ATGGCTTTCAGCCGACCAACGGCGTGGGCTATCAGCCGTATCGCGTGGTG 
               
               
                   
                 GTGCTGAGCTTTGAACTGCTGCATGCGCCGGCGACCGTGTGCGGCCCGAA 
               
               
                   
                 AAAAAGCACCAACCTGGTGAAAAACAAATGCGTGAACTTTAACTTTAACG 
               
               
                   
                 GCCTGACCGGCACCGGCGTGCTGACCGAAAGCAACAAAAAATTTCTGCCG 
               
               
                   
                 TTTCAGCAGTTTGGCCGCGATATTGCGGATACCACCGATGCGGTGCGCGA 
               
               
                   
                 TCCGCAGACCCTGGAAATTCTGGATATTACCCCGTGCAGCTTTGGCGGCG 
               
               
                   
                 TGAGCGTGATTACCCCGGGCACCAACACCAGCAACCAGGTGGCGGTGCTG 
               
               
                   
                 TATCAGGGCGTGAACTGCACCGAAGTGCCGGTGGCGATTCATGCGGATCA 
               
               
                   
                 GCTGACCCCGACCTGGCGCGTGTATAGCACCGGCAGCAACGTGTTTCAGA 
               
               
                   
                 CCCGCGCGGGCTGCCTGATTGGCGCGGAACATGTGAACAACAGCTATGAA 
               
               
                   
                 TGCGATATTCCGATTGGCGCGGGCATTTGCGCGAGCTATCAGACCCAGAC 
               
               
                   
                 CAACAGCCCGGGCAGCGCGAGCAGCGTGGCGAGCCAGAGCATTATTGCGT 
               
               
                   
                 ATACCATGAGCCTGGGCGCGGAAAACAGCGTGGCGTATAGCAACAACAGC 
               
               
                   
                 ATTGCGATTCCGACCAACTTTACCATTAGCGTGACCACCGAAATTCTGCC 
               
               
                   
                 GGTGAGCATGACCAAAACCAGCGTGGATTGCACCATGTATATTTGCGGCG 
               
               
                   
                 ATAGCACCGAATGCAGCAACCTGCTGCTGCAGTATGGCAGCTTTTGCACC 
               
               
                   
                 CAGCTGAACCGCGCGCTGACCGGCATTGCGGTGGAACAGGATAAAAACAC 
               
               
                   
                 CCAGGAAGTGTTTGCGCAGGTGAAACAGATTTATAAAACCCCGCCGATTA 
               
               
                   
                 AAGATTTTGGCGGCTTTAACTTTAGCCAGATTCTGCCGGATCCGAGCAAA 
               
               
                   
                 CCGAGCAAACGCAGCTTTATTGAAGATCTGCTGTTTAACAAAGTGACCCT 
               
               
                   
                 GGCGGATGCGGGCTTTATTAAACAGTATGGCGATTGCCTGGGCGATATTG 
               
               
                   
                 CGGCGCGCGATCTGATTTGCGCGCAGAAATTTAACGGCCTGACCGTGCTG 
               
               
                   
                 CCGCCGCTGCTGACCGATGAAATGATTGCGCAGTATACCAGCGCGCTGCT 
               
               
                   
                 GGCGGGCACCATTACCAGCGGCTGGACCTTTGGCGCGGGCGCGGCGCTGC 
               
               
                   
                 AGATTCCGTTTGCGATGCAGATGGCGTATCGCTTTAACGGCATTGGCGTG 
               
               
                   
                 ACCCAGAACGTGCTGTATGAAAACCAGAAACTGATTGCGAACCAGTTTAA 
               
               
                   
                 CAGCGCGATTGGCAAAATTCAGGATAGCCTGAGCAGCACCGCGAGCGCGC 
               
               
                   
                 TGGGCAAACTGCAGGATGTGGTGAACCAGAACGCGCAGGCGCTGAACACC 
               
               
                   
                 CTGGTGAAACAGCTGAGCAGCAACTTTGGCGCGATTAGCAGCGTGCTGAA 
               
               
                   
                 CGATATTCTGAGCCGCCTGGATCCGCCGGAAGCGGAAGTGCAGATTGATC 
               
               
                   
                 GCCTGATTACCGGCCGCCTGCAGAGCCTGCAGACCTATGTGACCCAGCAG 
               
               
                   
                 CTGATTCGCGCGGCGGAAATTCGCGCGAGCGCGAACCTGGCGGCGACCAA 
               
               
                   
                 AATGAGCGAATGCGTGCTGGGCCAGAGCAAACGCGTGGATTTTTGCGGCA 
               
               
                   
                 AAGGCTATCATCTGATGAGCTTTCCGCAGAGCGCGCCGCATGGCGTGGTG 
               
               
                   
                 TTTCTGCATGTGACCTATGTGCCGGCGCAGGAAAAAAACTTTACCACCGC 
               
               
                   
                 GCCGGCGATTTGCCATGATGGCAAAGCGCATTTTCCGCGCGAAGGCGTGT 
               
               
                   
                 TTGTGAGCAACGGCACCCATTGGTTTGTGACCCAGCGCAACTTTTATGAA 
               
               
                   
                 CCGCAGATTATTACCACCGATAACACCTTTGTGAGCGGCAACTGCGATGT 
               
               
                   
                 GGTGATTGGCATTGTGAACAACACCGTGTATGATCCGCTGCAGCCGGAAC 
               
               
                   
                 TGGATAGCTTTAAAGAAGAACTGGATAAATATTTTAAAAACCATACCAGC 
               
               
                   
                 CCGGATGTGGATCTGGGCGATATTAGCGGCATTAACGCGAGCGTGGTGAA 
               
               
                   
                 CATTCAGAAAGAAATTGATCGCCTGAACGAAGTGGCGAAAAACCTGAACG 
               
               
                   
                 AAAGCCTGATTGATCTGCAGGAACTGGGCAAATATGAACAGGGCTATATT 
               
               
                   
                 CCGGAAGCGCCGCGCGATGGCCAGGCGTATGTGCGCAAAGATGGCGAATG 
               
               
                   
                 GGTGCTGCTGAGCACCTTTCTGGCATTAGCCTATGGAAGTCAGGGTAATC 
               
               
                   
                 TTAATCCATTAATTAATGAAATCAGCAAAATCATTTCAGCTGCAGGTAAT 
               
               
                   
                 TTTGATGTTAAAGAGGAAAGAGCGGCCGCGTCTTTATTGCAGTTGTCCGG 
               
               
                   
                 TAATGCCAGTGATTTTTCATATGGACGGAACTCAATAACTTTGACAGCAT 
               
               
                   
                 CAGCATAAGAGCTCtttattaatttaaataatagcaatcttactgggctg 
               
               
                   
                 tgccacataagattgctatttttttggagtcataatggattcttgtcata 
               
               
                   
                 aaattgattatgggttatacgccctggagattttagcccaataccataac 
               
               
                   
                 gtctctgttaacccggaagaaattaaacatagatttgacacagacgggac 
               
               
                   
                 tggtctgggattaacgtcatggttgcttgctgcgaaatctttagaactaa 
               
               
                   
                 aggtaaaacaggtaaaaaaaacaattgaccgattaaactttatttctctg 
               
               
                   
                 cccgcattagtctggagagaggatggatgtcattttattctgactaaagt 
               
               
                   
                 cagtaaagaagcaaacagatatcttatttttgatctggagcagcgaaatc 
               
               
                   
                 cccgtgttctcgaacagtctgagtttgaggcgttatatcaggggcatatt 
               
               
                   
                 attcttattgcttcccgttcttctgttaccgggaaactggcaaaatttga 
               
               
                   
                 ctttacctggtttattcctgccattataaaatacaggaaaatatttattg 
               
               
                   
                 aaacccttgttgtatctgtttttttacaattatttgcattaataaccccc 
               
               
                   
                 cttttttttcaggtggttatggacaaagtattagtacacagggggttttc 
               
               
                   
                 aacccttaatgttattactgtcgcattatctgttgtggtggtgtttgaga 
               
               
                   
                 ttatactcagcggtttaagaacttacatttttgcacatagtacaagtcgg 
               
               
                   
                 attgatgttgagttgggtgccaaactcttccggcatttactggcgctacc 
               
               
                   
                 gatctcttattttgagagtcgtcgtgttggtgatactgttgccagggtaa 
               
               
                   
                 gagaattagaccagatccgtaatttcctgacaggacaggcattaacatct 
               
               
                   
                 gttctggacttattattttcattcatattttttgcggtaatgtggtatta 
               
               
                   
                 cagcccaaagcttactctggtgatcttattttcgctgccctgttatgctg 
               
               
                   
                 catggtctgtttttattagccccattttgcgacgtcgccttgatgataag 
               
               
                   
                 ttttcacggaatgcggataatcaatctttcctggtggaatcagtcacggc 
               
               
                   
                 gattaacactataaaagctatggcagtctcacctcagatgacgaacatat 
               
               
                   
                 gggacaaacaattggcaggatatgttgctgcaggctttaaagtgacagta 
               
               
                   
                 ttagccaccattggtcaacaaggaatacagttaatacaaaagactgttat 
               
               
                   
                 gatcatcaacctgtggttgggtgcacacctggttatttccggggatttaa 
               
               
                   
                 gtattggtcagttaattgcttttaatatgcttgcaggtcagattgttgca 
               
               
                   
                 ccggttattcgccttgcacaaatctggcaggatttccagcaggttggtat 
               
               
                   
                 atcagttacccgccttggtgatgtgcttaactctccaactgaaagttatc 
               
               
                   
                 atgggaaactggcattaccggaaattaatggtgatatcacttttcgtaat 
               
               
                   
                 atccggtttcgctataagcctgactctccggttattttagataatatcaa 
               
               
                   
                 tctcagtattaagcagggggaggttattggtattgtcggacgttctggtt 
               
               
                   
                 caggaaaaagcacattaactaaattaattcaacgtttttatattcctgaa 
               
               
                   
                 aatggccaggtcttaattgatggacatgatcttgcgttggccgatcctaa 
               
               
                   
                 ctggttacgtcgtcaggtgggggttgtgttgcaggacaatgtgctgctta 
               
               
                   
                 atcgcagtattattgataatatttcactggctaatcctggcatgtccgtc 
               
               
                   
                 gaaaaagttatttatgcagcgaaattagcaggtgctcatgattttatttc 
               
               
                   
                 tgaattgcgtgaggggtataacaccattgtcggggaacagggggcaggat 
               
               
                   
                 tatccggaggtcaacgtcaacgcatcgcaattgcaagggcgctggtgaac 
               
               
                   
                 aaccctaaaatactcatctttgatgaagcaaccagtgctctggattatga 
               
               
                   
                 gtcggagcatgtcatcatgcgcaatatgcacaaaatatgtaagggcagaa 
               
               
                   
                 cggttataatcattgctcatcgtctgtctacagtaaaaaatgcagaccgc 
               
               
                   
                 attattgtcatggaaaaagggaaaattgttgaacagggtaaacataagga 
               
               
                   
                 gctgctttctgaaccggaaagtttatacagttacttatatcagttacagt 
               
               
                   
                 cagactaacagaaagaacagaagaatatgaaaacatggttaatggggttc 
               
               
                   
                 agcgagttcctgttgcgctataaacttgtctggagtgaaacatggaaaat 
               
               
                   
                 ccggaagcagttagatactccggtacgtgaaaaggacgaaaatgaattct 
               
               
                   
                 tacccgctcatctggaattaattgaaacgccggtatccagacggccgcgt 
               
               
                   
                 ctggttgcttattttattatggggtttctggttattgctttcattttatc 
               
               
                   
                 tgttttaggccaggtggaaattgttgccactgcaaatgggaaattaacac 
               
               
                   
                 tcagtgggcgtagcaaagaaattaaacctattgaaaactcgatagttaaa 
               
               
                   
                 gaaattatcgtaaaagaaggagagtcagtccggaaaggggatgtgttatt 
               
               
                   
                 aaagcttacagcgctgggagctgaagctgatacgttaaaaacgcagtcat 
               
               
                   
                 cactgttacaggccaggctggaacaaattcggtatcaaattctgagccgg 
               
               
                   
                 tcaattgaattaaataaacttcctgaactgaaacttcctgatgagcctta 
               
               
                   
                 ttttcagaatgtatctgaagaggaagtactgcgtttaacttctttgataa 
               
               
                   
                 aagaacagttttccacatggcaaaatcagaagtatcaaaaagaactgaat 
               
               
                   
                 ctggataagaaaagagcagagcgattaacaatacttgcccgtataaaccg 
               
               
                   
                 ttatgaaaatgtatcgagggttgaaaaaagccgtctggatgatttcagga 
               
               
                   
                 gcctgttgcataaacaggcaattgcaaaacatgctgtacttgagcaggag 
               
               
                   
                 aataaatatgttgaggcagcaaatgaattacgggtttataaatcgcaact 
               
               
                   
                 ggagcaaattgagagtgagatattgtctgcaaaagaagaatatcagcttg 
               
               
                   
                 tcacgcagctttttaaaaatgaaattttagacaagctaagacaaacaaca 
               
               
                   
                 gacagcattgagttattaactctggagttagagaaaaatgaagagcgtca 
               
               
                   
                 acaggcttcagtaatcagggcccctgtttcgggaaaagttcagcaactga 
               
               
                   
                 aggttcatactgaaggtggggttgttacaacagcggaaacactgatggtc 
               
               
                   
                 atcgttccggaagatgacacgctggaggttactgctctggtacaaaataa 
               
               
                   
                 agatattggttttattaacgtcgggcagaatgccatcattaaagtggagg 
               
               
                   
                 cttttccttacacccgatatggttatctggtgggtaaggtaaaaaatata 
               
               
                   
                 aatttagatgcaatagaagaccagaaactgggactcgtttttaatgtcat 
               
               
                   
                 tgtttctgttgaagagaatgatttgtcaaccgggaataagcacattccat 
               
               
                   
                 taagctcgggtatggctgtcactgcagaaataaagactggaatgcgaagc 
               
               
                   
                 gtaatcagttatcttcttagtcccctggaagaatctgtaacagaaagttt 
               
               
                   
                 acatgagcgttaagtctcagagcagcggtatccggctcatatcttctcct 
               
               
                   
                 gtcagatccaagggcgaattccagcacactggcggccgttactagtggat 
               
               
                   
                 ccaccggtctcgaggggcgcgccggtaccgaattctaattaatgggcatc 
               
               
                   
                 atttttcagctatttcatttataaaataagttatgaaaaaaatcatcata 
               
               
                   
                 ttactaacgacatttttcctgctttcgggatgcactattcccagggcggt 
               
               
                   
                 atttaaatccagccttattaatcaggacgatcctcgttataatctggtcg 
               
               
                   
                 aagtcacgccgacattaaaactaagcgctcccgatactgtgccgaaaact 
               
               
                   
                 attgtcgatccggtttttgccgcaaataactggcactggacatctttggc 
               
               
                   
                 taaaggcgatgtgctgcatatcactattttatcctcgggcggggctggat 
               
               
                   
                 atttatccaataacgcgagcggcgaccgtgcggattttgaaaatattctt 
               
               
                   
                 gtgactgacagtaataccgttcaggtgccttatgccgggacaatcccggt 
               
               
                   
                 ttctggattggatgtgacgcaactggctgatgagatcaaaaagcgacttt 
               
               
                   
                 cgcgcgttgtcctgaatcctcaggtgattgtgacacttaccgcccgcacc 
               
               
                   
                 ggagccatggtgacggtcgagggcagcgggaaaacgggtcgataccctct 
               
               
                   
                 cgaacagagtatgaatcgtctgagtcatcttttggcaacggcagtggcag 
               
               
                   
                 tagaaaataccagcacagatatgatggaagttcacgtgacgcgccagcag 
               
               
                   
                 cattatttcactgcgcgactgtcagatatttatcagtatcccggattgga 
               
               
                   
                 tattgccttacagccggatgaccgtatcactctgcgtcaagtgaccgagt 
               
               
                   
                 atgttaatgtgctcggcgcagcaggtgttcaggggaaacatgctctggtt 
               
               
                   
                 cagcgtcattccagcgtcgtggatgctttggcgctcgcgaaaggattaaa 
               
               
                   
                 tgacaatcttgccgatccgcaagcgatttttctgtacaagcataacgaag 
               
               
                   
                 cggaacaggcgaaacagcagatgcgtaagctgaatatctatcatgtcgat 
               
               
                   
                 atgagccaacctaactcggtgtttttagcgcaggccatacgagtggacaa 
               
               
                   
                 cggcgatgttatctatatctcgaacgcctctttgactgatttcgctaagg 
               
               
                   
                 tgaaagccgcattcgatagctttttgac 
               
               
                   
               
               
                 SEQ ID NO: 9 (Ty21a- 
                 cccgttcctcattgatttgattgctaacgtcatgagcattactctatcat 
               
               
                 M-N-AR Construct) 
                 tccagaatgcctcaatgaacaagttgtttgagaaagagtgtcgcaatgtt 
               
               
                   
                 gcaaccagagcccttaaatatgtacgccagaagaaaactgaagggcgtct 
               
               
                   
                 ggatgaagcattgtctgtattgattagcctgaaacgaattgagcctgatg 
               
               
                   
                 tttctcgtctgatgcgtgaatataagcaaattatcagattatttaatgag 
               
               
                   
                 tcacggaaggatggcggtagcactatcacgtcttatgaacatctagacta 
               
               
                   
                 tgcgaaaaaattactcgtttttgatagcgaaaatgcctatgccttgaaat 
               
               
                   
                 atgccgcattaaatgcaatgcatttacgcgactacacgcaggctttgcag 
               
               
                   
                 tattggcagcgactggagaaagtgaatggaccaacggagccggtgacaag 
               
               
                   
                 gcagatctcgacctgcataaccgcattacaaaaaaatacatcagggaagt 
               
               
                   
                 cgtaacccggggtgtaggctggagctgcttcgaagttcctatactttcta 
               
               
                   
                 gagaataggaacttcggaataggaacttcaagatcccctcacgctgccgc 
               
               
                   
                 aagcactcagggcgcaagggctgctaaaggaagcggaacacgtagaaagc 
               
               
                   
                 cagtccgcagaaacggtgctgaccccggatgaatgtcagctactgggcta 
               
               
                   
                 tctggacaagggaaaacgcaagcgcaaagagaaagcaggtagcttgcagt 
               
               
                   
                 gggcttacatggcgatagctagactgggcggttttatggacagcaagcga 
               
               
                   
                 accggaattgccagctggggcgccctctggtaaggttgggaagccctgca 
               
               
                   
                 aagtaaactggatggctttcttgccgccaaggatctgatggcgcagggga 
               
               
                   
                 tcaagatctgatcaagagacaggatgaggatcgtttcgcatgattgaaca 
               
               
                   
                 agatggattgcacgcaggttctccggccgcttgggtggagaggctattcg 
               
               
                   
                 gctatgactgggcacaacagacaatcggctgctctgatgccgccgtgttc 
               
               
                   
                 cggctgtcagcgcaggggcgcccggttctttttgtcaagaccgacctgtc 
               
               
                   
                 cggtgccctgaatgaactgcaggacgaggcagcgcggctatcgtggctgg 
               
               
                   
                 ccacgacgggcgttccttgcgcagctgtgctcgacgttgtcactgaagcg 
               
               
                   
                 ggaagggactggctgctattgggcgaagtgccggggcaggatctcctgtc 
               
               
                   
                 atctcaccttgctcctgccgagaaagtatccatcatggctgatgcaatgc 
               
               
                   
                 ggcggctgcatacgcttgatccggctacctgcccattcgaccaccaagcg 
               
               
                   
                 aaacatcgcatcgagcgagcacgtactcggatggaagccggtcttgtcga 
               
               
                   
                 tcaggatgatctggacgaagagcatcaggggctcgcgccagccgaactgt 
               
               
                   
                 tcgccaggctcaaggcgcgcatgcccgacggcgaggatctcgtcgtgacc 
               
               
                   
                 catggcgatgcctgcttgccgaatatcatggtggaaaatggccgcttttc 
               
               
                   
                 tggattcatcgactgtggccggctgggtgtggcggaccgctatcaggaca 
               
               
                   
                 tagcgttggctacccgtgatattgctgaagagcttggcggcgaatgggct 
               
               
                   
                 gaccgcttcctcgtgctttacggtatcgccgctcccgattcgcagcgcat 
               
               
                   
                 cgccttctatcgccttcttgacgagttcttctgagcgggactctggggtt 
               
               
                   
                 cgaaatgaccgaccaagcgacgcccaacctgccatcacgagatttcgatt 
               
               
                   
                 ccaccgccgccttctatgaaaggttgggcttcggaatcgttttccgggac 
               
               
                   
                 gccggctggatgatcctccagcgcggggatctcatgctggagttcttcgc 
               
               
                   
                 ccaccccagcttcaaaagcgctctgaagttcctatactttctagagaata 
               
               
                   
                 ggaacttcggaataggaactaaggaggatattcatatatgcattaatgtc 
               
               
                   
                 taacaattcgttcaagccgacgccgcttcgcggcgcggcttaactcaagc 
               
               
                   
                 gttagatgcactaagcacataattgctcacagccaaactatcaggtcaag 
               
               
                   
                 tctgcttttattatttttaagcgtgcataataagccctacacaaattggg 
               
               
                   
                 agatatatcatgaaaggctggctttttcttgttatcgcaatagttgCGAT 
               
               
                   
                 CGTTATGACAACTTGACGGCTACATCATTCACTTTTTCTTCACAACCGGC 
               
               
                   
                 ACGGAACTCGCTCGGGCTGGCCCCGGTGCATTTTTTAAATACCCGCGAGA 
               
               
                   
                 AGTAGAGTTGATCGTCAAAACCAACATTGCGACCGACGGTGGCGATAGGC 
               
               
                   
                 ATCCGGGTGGTGCTCAAAAGCAGCTTCGCCTGGCTGATACGTTGGTCCTC 
               
               
                   
                 GCGCCAGCTTAAGACGCTAATCCCTAACTGCTGGCGGAAAAGATGTGACA 
               
               
                   
                 GACGCGACGGCGACAAGCAAACATGCTGTGCGACGCTGGCGATATCAAAA 
               
               
                   
                 TTGCTGTCTGCCAGGTGATCGCTGATGTACTGACAAGCCTCGCGTACCCG 
               
               
                   
                 ATTATCCATCGGTGGATGGAGCGACTCGTTAATCGCTTCCATGCGCCGCA 
               
               
                   
                 GTAACAATTGCTCAAGCAGATTTATCGCCAGCAGCTCCGAATAGCGCCCT 
               
               
                   
                 TCCCCTTGCCCGGCGTTAATGATTTGCCCAAACAGGTCGCTGAAATGCGG 
               
               
                   
                 CTGGTGCGCTTCATCCGGGCGAAAGAACCCCGTATTGGCAAATATTGACG 
               
               
                   
                 GCCAGTTAAGCCATTCATGCCAGTAGGCGCGCGGACGAAAGTAAACCCAC 
               
               
                   
                 TGGTGATACCATTCGCGAGCCTCCGGATGACGACCGTAGTGATGAATCTC 
               
               
                   
                 TCCTGGCGGGAACAGCAAAATAACACCCGGTCGGCAAACAAATTCTCGTC 
               
               
                   
                 CCTGATTTTTCACCACCCCCTGACCGCGAATGGTGAGATTGAGAATATAA 
               
               
                   
                 CCTTTCATTCCCAGCGGTCGGTCGATAAAAAAATCGAGATAACCGTTGGC 
               
               
                   
                 CTCAATCGGCGTTAAACCCGCCACCAGATGGGCATTAAACGAGTATCCCG 
               
               
                   
                 GCAGCAGGGGATCATTTTGCGCTTCAGCCATACTTTTCATACTCCCGCCA 
               
               
                   
                 TTCAGAGAAGAAACCAATTGTCCATATTGCATCAGACATTGCCGTCACTG 
               
               
                   
                 CGTCTTTTACTGGCTCTTCTCGCTAACCAAACCGGTAACCCCGCTTATTA 
               
               
                   
                 AAAGCATTCTGTAACAAAGCGGGACCAAAGCCATGACAAAAACGCGTAAC 
               
               
                   
                 AAAAGTGTCTATAATCACGGCAGAAAAGTCCACATTGATTATTTGCACGG 
               
               
                   
                 CGTCACACTTTGCTATGCCATAGCATTTTTATCCATAAGATTAGCGGATC 
               
               
                   
                 CTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACCCGTTT 
               
               
                   
                 TTTTGGGAATTCGAGCTCGGAGTTAACAAAAGATGTTAGATGCAAACAAA 
               
               
                   
                 TTACAGCAGGCAGTGGATCAGGCTTACACCCAATTTCACTCACTTAACGG 
               
               
                   
                 CGGACAAAATGCCGATTACATTCCCTTTCTGGCGAATGTACCAGGTCAAC 
               
               
                   
                 TGGCGGCAGTGGCTATCGTGACNTGCGATGGCAACGTCTATAGTGCGGGT 
               
               
                   
                 GACAGTGATTACCGCTTTGCACTGGAATCCATCTCTAAAGTCTGTACGTT 
               
               
                   
                 AGCCCTTGCGTTAGAAGATGTCGGCCCGCAGGCGGTACAGGACAAAATTG 
               
               
                   
                 GCGCTGACCCGACCGGATTGCCCTTTAACTCAGTTATCGCCTTAGAGTTG 
               
               
                   
                 CATGGCGGCAAACCGCTGTCGCCACTGGTAAATGCTGGCGCTATTGCCAC 
               
               
                   
                 CACCAGCCTGATTAACGCTGAAAATGTTGAACAACGCTGGCAGCGAATTT 
               
               
                   
                 TACATATCCAACAGCAACTGGCTGGTGAGCAGGTAGCGCTCTCTGACGAA 
               
               
                   
                 GTCAACCAGTCGGAACAAACAACCAACTTCCATAACCGGGCCATTGCCTG 
               
               
                   
                 GCTGCTGTACTCCGCCGGATATCTCTATTGTGATGCAATGGAAGCCTGTG 
               
               
                   
                 ACGTGTACACCCGTCAGTGCTCTACGCTCATCAATACTGTTGAACTGGCA 
               
               
                   
                 ACGCTTGGCGCGACGCTGGCGGCGGGTGGTGTGAATCCGTTGACGCATAA 
               
               
                   
                 ACGCGTTCTTCAGGCCGACAACGTGCCGTACATTCTGGCCGAAATGATGA 
               
               
                   
                 TGGAAGGGCTGTATGGTCGCTCCGGTGACTGGGCGTATCGCGTTGGTTTA 
               
               
                   
                 CCGGGCAAAAGCGGTGTAGGTGGCGGTATTCTGGCGGTCGTCCCTGGCGT 
               
               
                   
                 GATGGGAATTGCCGCGTTCTCACCACCGCTGGACGAAGAAGGCAACAGTG 
               
               
                   
                 TTCGCGGTCAAAAAATGGTGGCATCGGTCGCTAAGCAACTCGGCTATAAC 
               
               
                   
                 GTGTTTAAGGGCTGACCATGGAGTGTTTTAATGCGATCCAATCATTTTAA 
               
               
                   
                 GGAGTTTAAAATGGATAAGAAGCAAGTAACGGATTTAAGGTCGGAACTAC 
               
               
                   
                 TCGATTCACGTTTTGGTGCGAAGTCTATTTCCACTATCGCAGAATCAAAA 
               
               
                   
                 CGTTTTCCGCTGCACGAAATGCGCGACGATGTCGCATTTCAGATTATCAA 
               
               
                   
                 TGATGAATTATATCTTGATGGCAACGCTCGTCAGAACCTGGCCACTTTCT 
               
               
                   
                 GCCAGACCTGGGACGACGAAAACGTCCACAAGTTGATGGATTTATCCATT 
               
               
                   
                 AACAAAAACTGGATCGACAAAGAAGAATATCCGCAATCCGCAGCCATCGA 
               
               
                   
                 CCTGCGTTGCGTAAACATGGTTGCCGATCTGTGGCATGCGCCTGCGCCGA 
               
               
                   
                 AAAATGGTCAGGCCGTTGGCACCAACACCATTGGTTCTTCCGAGGCCTGT 
               
               
                   
                 ATGCTCGGCGGGATGGCGATGAAATGGCGTTGGCGCAAGCGTATGGAAGC 
               
               
                   
                 TGCAGGCAAACCAACGGATAAACCAAACCTGGTGTGCGGTCCGGTGCAAA 
               
               
                   
                 TCTGCTGGCATAAATTCGCCCGCTACTGGGATGTGGAGCTGCGTGAGATC 
               
               
                   
                 CCTATGCGCCCCGGTCAGTTGTTTATGGACCCGAAACGCATGATTGAAGC 
               
               
                   
                 CTGCGACGAAAATACCATCGGCGTGGTGCCGACTTTCGGCGTGACCTACA 
               
               
                   
                 CCGGTAACTATGAGTTCCCGCAACCGCTGCACGATGCACTGGATAAATTC 
               
               
                   
                 CAGGCCGACACCGGTATCGACATCGACATGCACATCGACGCCGCCAGCGG 
               
               
                   
                 TGGCTTTCTGGCACCGTTCGTCGCCCCGGATATCGTCTGGGACTTCCGCC 
               
               
                   
                 TGCCGCGTGTGAAATCGATCAGTGCTTCAGGCCATAAATTCGGTCTGGCT 
               
               
                   
                 CCGCTGGGCTGCGGCTGGGTTATCTGGCGTGACGAAGAAGCGCTGCCGCA 
               
               
                   
                 GGAACTGGTGTTCAACGTTGACTACCTCGGTGGTCAGATTGGTACTTTTG 
               
               
                   
                 CCATCAACTTCTCCCGCCCGGCGGGTCAGGTGATTGCACAGTACTATGAA 
               
               
                   
                 TTCCTGCGCCTTGGTCGTGAAGGCTATACCAAAGTACAGAATGCTTCCTA 
               
               
                   
                 CCAGGTCGCTGCCTATCTGGCGGATGAGATCGCCAAACTGGGGCCGTATG 
               
               
                   
                 AGTTCATCTGTACCGGTCGCCCGGACGAAGGCATCCCGGCGGTTTGCTTC 
               
               
                   
                 AAACTGAAAGATGGTGAAGATCCGGGATACACCCTGTACGACCTCTCTGA 
               
               
                   
                 ACGTCTGCGTCTGCGCGGCTGGCAGGTTCCGGCCTTCACTCTCGGCGGTG 
               
               
                   
                 AAGCCACCGACATCGTGGTGATGCGCATTATGTGTCGTCGCGGCTTCGAA 
               
               
                   
                 ATGGACTTTGCTGAACTGTTGCTGGAAGACTACAAAGCCTCCCTGAAATA 
               
               
                   
                 TCTCAGCGATCACCCGAAACTGCAGGGTATTGCCCAGCAGAACAGCTTTA 
               
               
                   
                 AACATACCTGATAACCGTTTTGGAGTGATAATATGGCTACATCAGTACAG 
               
               
                   
                 ACAGGTAAAGCTAAGCAGCTCACATTACTCGGATTCTTTGCCATAACGGC 
               
               
                   
                 ATCGATGGTAATGGCTGTTTATGAATACCCTACCTTCGCAACATCGGGCT 
               
               
                   
                 TTTCATTAGTCTTCTTCCTGCTATTAGGCGGGATTTTATGGTTTATTCCC 
               
               
                   
                 GTGGGACTTTGTGCTGCGGAAATGGCCACCGTCGACGGCTGGGAAGAAGG 
               
               
                   
                 TGGTGTCTTCGCCTGGGTATCAAATACTCTGGGTCCGAGATGGGGATTTG 
               
               
                   
                 CAGCGATCTCATTTGGCTATCTGCAAATCGCCATTGGTTTTATTCCGATG 
               
               
                   
                 CTCTATTTCGTGTTAGGGGCACTCTCCTACATCCTGAAATGGCCAGCGCT 
               
               
                   
                 GAATGAAGACCCCATTACCAAAACTATTGCAGCACTCATCATTCTTTGGG 
               
               
                   
                 CGCTGGCATTAACGCAGTTTGGTGGCACGAAATACACGGCGCGAATTGCT 
               
               
                   
                 AAAGTTGGCTTCTTCGCCGGTATCCTGTTACCTGCATTTATTTTGATCGC 
               
               
                   
                 ATTAGCGGCTATTTACCTGCACTCCGGTGCCCCCGTTGCTATCGAAATGG 
               
               
                   
                 ATTCGAAGACCTTCTTCCCTGACTTCTCTAAAGTGGGCACACTGGTTGTA 
               
               
                   
                 TTTGTTGCCTTCATTTTGAGTTATATGGGCGTAGAAGCTTCCGCAACTCA 
               
               
                   
                 CGTCAATGAAATGAGCAACCCCGGGCGCGACTATCCACTGGCTATGTTAC 
               
               
                   
                 TGCTGATGGTGGCGGCAATCTGCTTAAGCTCTGTTGGCGGTTTGTCTATT 
               
               
                   
                 GCGATGGTCATTCCGGGTAATGAAATCAACCTCTCCGCAGGGGTAATGCA 
               
               
                   
                 AACCTTTACCGTTCTGATGTCCCATGTGGCACCGGAAATTGAGTGGACGG 
               
               
                   
                 TTCGCGTGATCTCCGCACTGCTGTTGCTGGGTGTTCTGGCGGAAATCGCC 
               
               
                   
                 TCCTGGATTGTTGGTCCTTCTCGCGGGATGTATGTCACAGCGCAGAAAAA 
               
               
                   
                 CCTGCTGCCAGCGGCATTCGCTAAAATGAACAAAAATGGCGTACCGGTAA 
               
               
                   
                 CGCTGGTCATTTCGCAGCTGGTGATTACTTCTATCGCGTTGATCATCCTC 
               
               
                   
                 ACCAATACCGGTGGCGGTAACAACATGTCCTTCCTGATCGCACTGGCGCT 
               
               
                   
                 GACGGTGGTGATTTATCTGTGTGCTTATTTCATGCTGTTTATTGGCTACA 
               
               
                   
                 TTGTGTTGGTTCTTAAACATCCTGACTTAAAACGCACATTTAATATCCCT 
               
               
                   
                 GGTGGTAAAGGGGTGAAACTGGTCGTGGCAATTGTCGGTCTGCTGACTTC 
               
               
                   
                 AATTATGGCGTTTATTGTTTCCTTCCTGCCGCCGGATAACATCCAGGGTG 
               
               
                   
                 ATTCTACCGATATGTATGTTGAATTACTGGTTGTTAGTTTCCTGGTGGTA 
               
               
                   
                 CTTGCCCTGCCCTTTATTCTCTATGCTGTTCATGATCGTAAAGGCAAAGC 
               
               
                   
                 GAATACCGGCGTCACTCTGGAGCCAATCAACAGTCAGAACGCACCAAAAG 
               
               
                   
                 GTCACTTCTTCCTGCACCCGCGTGCACGTTCACCACACTATATTGTGATG 
               
               
                   
                 AATGACAAGAAACACTAACGATCGgcgaagtaatcgcaacatccgcatta 
               
               
                   
                 aaatctagcgagggctttactgtcgaccaaaaaaatattgacaacataaa 
               
               
                   
                 aaactttgtgttatacttgtaacgGactctagagggtattaataatgaaa 
               
               
                   
                 ggagaggacatGAGCTCCCTAGGATGAGCGATAACGGCCCGCAGAACCAG 
               
               
                   
                 CGCAACGCGCCGCGCATTACCTTTGGCGGCCCGAGCGATAGCACCGGCAG 
               
               
                   
                 CAACCAGAACGGCGAACGCAGCGGCGCGCGCAGCAAACAGCGCCGCCCGC 
               
               
                   
                 AGGGCCTGCCGAACAACACCGCGAGCTGGTTTACCGCGCTGACCCAGCAT 
               
               
                   
                 GGCAAAGAAGATCTGAAATTTCCGCGCGGCCAGGGCGTGCCGATTAACAC 
               
               
                   
                 CAACAGCAGCCCGGATGATCAGATTGGCTATTATCGCCGCGCGACCCGCC 
               
               
                   
                 GCATTCGCGGCGGCGATGGCAAAATGAAAGATCTGAGCCCGCGCTGGTAT 
               
               
                   
                 TTTTATTATCTGGGCACCGGCCCGGAAGCGGGCCTGCCGTATGGCGCGAA 
               
               
                   
                 CAAAGATGGCATTATTTGGGTGGCGACCGAAGGCGCGCTGAACACCCCGA 
               
               
                   
                 AAGATCATATTGGCACCCGCAACCCGGCGAACAACGCGGCGATTGTGCTG 
               
               
                   
                 CAGCTGCCGCAGGGCACCACCCTGCCGAAAGGCTTTTATGCGGAAGGCAG 
               
               
                   
                 CCGCGGCGGCAGCCAGGCGAGCAGCCGCAGCAGCAGCCGCAGCCGCAACA 
               
               
                   
                 GCAGCCGCAACAGCACCCCGGGCAGCAGCCGCGGCACCAGCCCGGCGCGC 
               
               
                   
                 ATGGCGGGCAACGGCGGCGATGCGGCGCTGGCGCTGCTGCTGCTGGATCG 
               
               
                   
                 CCTGAACCAGCTGGAAAGCAAAATGAGCGGCAAAGGCCAGCAGCAGCAGG 
               
               
                   
                 GCCAGACCGTGACCAAAAAAAGCGCGGCGGAAGCGAGCAAAAAACCGCGC 
               
               
                   
                 CAGAAACGCACCGCGACCAAAGCGTATAACGTGACCCAGGCGTTTGGCCG 
               
               
                   
                 CCGCGGCCCGGAACAGACCCAGGGCAACTTTGGCGATCAGGAACTGATTC 
               
               
                   
                 GCCAGGGCACCGATTATAAACATTGGCCGCAGATTGCGCAGTTTGCGCCG 
               
               
                   
                 AGCGCGAGCGCGTTTTTTGGCATGAGCCGCATTGGCATGGAAGTGACCCC 
               
               
                   
                 GAGCGGCACCTGGCTGACCTATACCGGCGCGATTAAACTGGATGATAAAG 
               
               
                   
                 ATCCGAACTTTAAAGATCAGGTGATTCTGCTGAACAAACATATTGATGCG 
               
               
                   
                 TATAAAACCTTTCCGCCGACCGAACCGAAAAAAGATAAAAAAAAAAAAGC 
               
               
                   
                 GGATGAAACCCAGGCGCTGCCGCAGCGCCAGAAAAAACAGCAGACCGTGA 
               
               
                   
                 CCCTGCTGCCGGCGGCGGATCTGGATGATTTTAGCAAACAGCTGCAGCAG 
               
               
                   
                 AGCATGAGCAGCGCGGATAGCACCCAGGCGCCTAGGCTTAAGATGGCGGA 
               
               
                   
                 TAGCAACGGCACCATTACCGTGGAAGAACTGAAAAAACTGCTGGAACAGT 
               
               
                   
                 GGAACCTGGTGATTGGCTTTCTGTTTCTGACCTGGATTTGCCTGCTGCAG 
               
               
                   
                 TTTGCGTATGCGAACCGCAACCGCTTTCTGTATATTATTAAACTGATTTT 
               
               
                   
                 TCTGTGGCTGCTGTGGCCGGTGACCCTGGCGTGCTTTGTGCTGGCGGCGG 
               
               
                   
                 TGTATCGCATTAACTGGATTACCGGCGGCATTGCGATTGCGATGGCGTGC 
               
               
                   
                 CTGGTGGGCCTGATGTGGCTGAGCTATTTTATTGCGAGCTTTCGCCTGTT 
               
               
                   
                 TGCGCGCACCCGCAGCATGTGGAGCTTTAACCCGGAAACCAACATTCTGC 
               
               
                   
                 TGAACGTGCCGCTGCATGGCACCATTCTGACCCGCCCGCTGCTGGAAAGC 
               
               
                   
                 GAACTGGTGATTGGCGCGGTGATTCTGCGCGGCCATCTGCGCATTGCGGG 
               
               
                   
                 CCATCATCTGGGCCGCTGCGATATTAAAGATCTGCCGAAAGAAATTACCG 
               
               
                   
                 TGGCGACCAGCCGCACCCTGAGCTATTATAAACTGGGCGCGAGCCAGCGC 
               
               
                   
                 GTGGCGGGCGATAGCGGCTTTGCGGCGTATAGCCGCTATCGCATTGGCAA 
               
               
                   
                 CTATAAACTGAACACCGATCATAGCAGCAGCAGCGATAACATTGCGCTGC 
               
               
                   
                 TGGTGCAGCTTAAGGCATTAGCCTATGGAAGTCAGGGTAATCTTAATCCA 
               
               
                   
                 TTAATTAATGAAATCAGCAAAATCATTTCAGCTGCAGGTAATTTTGATGT 
               
               
                   
                 TAAAGAGGAAAGAGCGGCCGCGTCTTTATTGCAGTTGTCCGGTAATGCCA 
               
               
                   
                 GTGATTTTTCATATGGACGGAACTCAATAACTTTGACAGCATCAGCATAA 
               
               
                   
                 GAGCTCtttattaatttaaataatagcaatcttactgggctgtgccacat 
               
               
                   
                 aagattgctatttttttggagtcataatggattcttgtcataaaattgat 
               
               
                   
                 tatgggttatacgccctggagattttagcccaataccataacgtctctgt 
               
               
                   
                 taacccggaagaaattaaacatagatttgacacagacgggactggtctgg 
               
               
                   
                 gattaacgtcatggttgcttgctgcgaaatctttagaactaaaggtaaaa 
               
               
                   
                 caggtaaaaaaaacaattgaccgattaaactttatttctctgcccgcatt 
               
               
                   
                 agtctggagagaggatggatgtcattttattctgactaaagtcagtaaag 
               
               
                   
                 aagcaaacagatatcttatttttgatctggagcagcgaaatccccgtgtt 
               
               
                   
                 ctcgaacagtctgagtttgaggcgttatatcaggggcatattattcttat 
               
               
                   
                 tgcttcccgttcttctgttaccgggaaactggcaaaatttgactttacct 
               
               
                   
                 ggtttattcctgccattataaaatacaggaaaatatttattgaaaccctt 
               
               
                   
                 gttgtatctgtttttttacaattatttgcattaataaccccccttttttt 
               
               
                   
                 tcaggtggttatggacaaagtattagtacacagggggttttcaaccctta 
               
               
                   
                 atgttattactgtcgcattatctgttgtggtggtgtttgagattatactc 
               
               
                   
                 agcggtttaagaacttacatttttgcacatagtacaagtcggattgatgt 
               
               
                   
                 tgagttgggtgccaaactcttccggcatttactggcgctaccgatctctt 
               
               
                   
                 attttgagagtcgtcgtgttggtgatactgttgccagggtaagagaatta 
               
               
                   
                 gaccagatccgtaatttcctgacaggacaggcattaacatctgttctgga 
               
               
                   
                 cttattattttcattcatattttttgcggtaatgtggtattacagcccaa 
               
               
                   
                 agcttactctggtgatcttattttcgctgccctgttatgctgcatggtct 
               
               
                   
                 gtttttattagccccattttgcgacgtcgccttgatgataagttttcacg 
               
               
                   
                 gaatgcggataatcaatctttcctggtggaatcagtcacggcgattaaca 
               
               
                   
                 ctataaaagctatggcagtctcacctcagatgacgaacatatgggacaaa 
               
               
                   
                 caattggcaggatatgttgctgcaggctttaaagtgacagtattagccac 
               
               
                   
                 cattggtcaacaaggaatacagttaatacaaaagactgttatgatcatca 
               
               
                   
                 acctgtggttgggtgcacacctggttatttccggggatttaagtattggt 
               
               
                   
                 cagttaattgcttttaatatgcttgcaggtcagattgttgcaccggttat 
               
               
                   
                 tcgccttgcacaaatctggcaggatttccagcaggttggtatatcagtta 
               
               
                   
                 cccgccttggtgatgtgcttaactctccaactgaaagttatcatgggaaa 
               
               
                   
                 ctggcattaccggaaattaatggtgatatcacttttcgtaatatccggtt 
               
               
                   
                 tcgctataagcctgactctccggttattttagataatatcaatctcagta 
               
               
                   
                 ttaagcagggggaggttattggtattgtcggacgttctggttcaggaaaa 
               
               
                   
                 agcacattaactaaattaattcaacgtttttatattcctgaaaatggcca 
               
               
                   
                 ggtcttaattgatggacatgatcttgcgttggccgatcctaactggttac 
               
               
                   
                 gtcgtcaggtgggggttgtgttgcaggacaatgtgctgcttaatcgcagt 
               
               
                   
                 attattgataatatttcactggctaatcctggcatgtccgtcgaaaaagt 
               
               
                   
                 tatttatgcagcgaaattagcaggtgctcatgattttatttctgaattgc 
               
               
                   
                 gtgaggggtataacaccattgtcggggaacagggggcaggattatccgga 
               
               
                   
                 ggtcaacgtcaacgcatcgcaattgcaagggcgctggtgaacaaccctaa 
               
               
                   
                 aatactcatctttgatgaagcaaccagtgctctggattatgagtcggagc 
               
               
                   
                 atgtcatcatgcgcaatatgcacaaaatatgtaagggcagaacggttata 
               
               
                   
                 atcattgctcatcgtctgtctacagtaaaaaatgcagaccgcattattgt 
               
               
                   
                 catggaaaaagggaaaattgttgaacagggtaaacataaggagctgcttt 
               
               
                   
                 ctgaaccggaaagtttatacagttacttatatcagttacagtcagactaa 
               
               
                   
                 cagaaagaacagaagaatatgaaaacatggttaatggggttcagcgagtt 
               
               
                   
                 cctgttgcgctataaacttgtctggagtgaaacatggaaaatccggaagc 
               
               
                   
                 agttagatactccggtacgtgaaaaggacgaaaatgaattcttacccgct 
               
               
                   
                 catctggaattaattgaaacgccggtatccagacggccgcgtctggttgc 
               
               
                   
                 ttattttattatggggtttctggttattgctttcattttatctgttttag 
               
               
                   
                 gccaggtggaaattgttgccactgcaaatgggaaattaacactcagtggg 
               
               
                   
                 cgtagcaaagaaattaaacctattgaaaactcgatagttaaagaaattat 
               
               
                   
                 cgtaaaagaaggagagtcagtccggaaaggggatgtgttattaaagctta 
               
               
                   
                 cagcgctgggagctgaagctgatacgttaaaaacgcagtcatcactgtta 
               
               
                   
                 caggccaggctggaacaaattcggtatcaaattctgagccggtcaattga 
               
               
                   
                 attaaataaacttcctgaactgaaacttcctgatgagccttattttcaga 
               
               
                   
                 atgtatctgaagaggaagtactgcgtttaacttctttgataaaagaacag 
               
               
                   
                 ttttccacatggcaaaatcagaagtatcaaaaagaactgaatctggataa 
               
               
                   
                 gaaaagagcagagcgattaacaatacttgcccgtataaaccgttatgaaa 
               
               
                   
                 atgtatcgagggttgaaaaaagccgtctggatgatttcaggagcctgttg 
               
               
                   
                 cataaacaggcaattgcaaaacatgctgtacttgagcaggagaataaata 
               
               
                   
                 tgttgaggcagcaaatgaattacgggtttataaatcgcaactggagcaaa 
               
               
                   
                 ttgagagtgagatattgtctgcaaaagaagaatatcagcttgtcacgcag 
               
               
                   
                 ctttttaaaaatgaaattttagacaagctaagacaaacaacagacagcat 
               
               
                   
                 tgagttattaactctggagttagagaaaaatgaagagcgtcaacaggctt 
               
               
                   
                 cagtaatcagggcccctgtttcgggaaaagttcagcaactgaaggttcat 
               
               
                   
                 actgaaggtggggttgttacaacagcggaaacactgatggtcatcgttcc 
               
               
                   
                 ggaagatgacacgctggaggttactgctctggtacaaaataaagatattg 
               
               
                   
                 gttttattaacgtcgggcagaatgccatcattaaagtggaggcttttcct 
               
               
                   
                 tacacccgatatggttatctggtgggtaaggtaaaaaatataaatttaga 
               
               
                   
                 tgcaatagaagaccagaaactgggactcgtttttaatgtcattgtttctg 
               
               
                   
                 ttgaagagaatgatttgtcaaccgggaataagcacattccattaagctcg 
               
               
                   
                 ggtatggctgtcactgcagaaataaagactggaatgcgaagcgtaatcag 
               
               
                   
                 ttatcttcttagtcccctggaagaatctgtaacagaaagtttacatgagc 
               
               
                   
                 gttaagtctcagagcagcggtatccggctcatatcttctcctgtcagatc 
               
               
                   
                 caagggcgaattccagcacactggcggccgttactagtggatccaccggt 
               
               
                   
                 ctcgaggggcgcgccggtaccgaattctaattaatgggcatcatttttca 
               
               
                   
                 gctatttcatttataaaataagttatgaaaaaaatcatcatattactaac 
               
               
                   
                 gacatttttcctgctttcgggatgcactattcccagggcggtatttaaat 
               
               
                   
                 ccagccttattaatcaggacgatcctcgttataatctggtcgaagtcacg 
               
               
                   
                 ccgacattaaaactaagcgctcccgatactgtgccgaaaactattgtcga 
               
               
                   
                 tccggtttttgccgcaaataactggcactggacatctttggctaaaggcg 
               
               
                   
                 atgtgctgcatatcactattttatcctcgggcggggctggatatttatcc 
               
               
                   
                 aataacgcgagcggcgaccgtgcggattttgaaaatattcttgtgactga 
               
               
                   
                 cagtaataccgttcaggtgccttatgccgggacaatcccggtttctggat 
               
               
                   
                 tggatgtgacgcaactggctgatgagatcaaaaagcgactttcgcgcgtt 
               
               
                   
                 gtcctgaatcctcaggtgattgtgacacttaccgcccgcaccggagccat 
               
               
                   
                 ggtgacggtcgagggcagcgggaaaacgggtcgataccctctcgaacaga 
               
               
                   
                 gtatgaatcgtctgagtcatcttttggcaacggcagtggcagtagaaaat 
               
               
                   
                 accagcacagatatgatggaagttcacgtgacgcgccagcagcattattt 
               
               
                   
                 cactgcgcgactgtcagatatttatcagtatcccggattggatattgcct 
               
               
                   
                 tacagccggatgaccgtatcactctgcgtcaagtgaccgagtatgttaat 
               
               
                   
                 gtgctcggcgcagcaggtgttcaggggaaacatgctctggttcagcgtca 
               
               
                   
                 ttccagcgtcgtggatgctttggcgctcgcgaaaggattaaatgacaatc 
               
               
                   
                 ttgccgatccgcaagcgatttttctgtacaagcataacgaagcggaacag 
               
               
                   
                 gcgaaacagcagatgcgtaagctgaatatctatcatgtcgatatgagcca 
               
               
                   
                 acctaactcggtgtttttagcgcaggccatacgagtggacaacggcgatg 
               
               
                   
                 ttatctatatctcgaacgcctctttgactgatttcgctaaggtgaaagcc 
               
               
                   
                 gcattcgatagctttttgac 
               
               
                   
               
               
                 SEQ ID NO: 10 (Ty21a- 
                 cccgttcctcattgatttgattgctaacgtcatgagcattactctatcat 
               
               
                 eS-M-N-AR Construct) 
                 tccagaatgcctcaatgaacaagttgtttgagaaagagtgtcgcaatgtt 
               
               
                   
                 gcaaccagagcccttaaatatgtacgccagaagaaaactgaagggcgtct 
               
               
                   
                 ggatgaagcattgtctgtattgattagcctgaaacgaattgagcctgatg 
               
               
                   
                 tttctcgtctgatgcgtgaatataagcaaattatcagattatttaatgag 
               
               
                   
                 tcacggaaggatggcggtagcactatcacgtcttatgaacatctagacta 
               
               
                   
                 tgcgaaaaaattactcgtttttgatagcgaaaatgcctatgccttgaaat 
               
               
                   
                 atgccgcattaaatgcaatgcatttacgcgactacacgcaggctttgcag 
               
               
                   
                 tattggcagcgactggagaaagtgaatggaccaacggagccggtgacaag 
               
               
                   
                 gcagatctcgacctgcataaccgcattacaaaaaaatacatcagggaagt 
               
               
                   
                 cgtaacccggggtgtaggctggagctgcttcgaagttcctatactttcta 
               
               
                   
                 gagaataggaacttcggaataggaacttcaagatcccctcacgctgccgc 
               
               
                   
                 aagcactcagggcgcaagggctgctaaaggaagcggaacacgtagaaagc 
               
               
                   
                 cagtccgcagaaacggtgctgaccccggatgaatgtcagctactgggcta 
               
               
                   
                 tctggacaagggaaaacgcaagcgcaaagagaaagcaggtagcttgcagt 
               
               
                   
                 gggcttacatggcgatagctagactgggcggttttatggacagcaagcga 
               
               
                   
                 accggaattgccagctggggcgccctctggtaaggttgggaagccctgca 
               
               
                   
                 aagtaaactggatggctttcttgccgccaaggatctgatggcgcagggga 
               
               
                   
                 tcaagatctgatcaagagacaggatgaggatcgtttcgcatgattgaaca 
               
               
                   
                 agatggattgcacgcaggttctccggccgcttgggtggagaggctattcg 
               
               
                   
                 gctatgactgggcacaacagacaatcggctgctctgatgccgccgtgttc 
               
               
                   
                 cggctgtcagcgcaggggcgcccggttctttttgtcaagaccgacctgtc 
               
               
                   
                 cggtgccctgaatgaactgcaggacgaggcagcgcggctatcgtggctgg 
               
               
                   
                 ccacgacgggcgttccttgcgcagctgtgctcgacgttgtcactgaagcg 
               
               
                   
                 ggaagggactggctgctattgggcgaagtgccggggcaggatctcctgtc 
               
               
                   
                 atctcaccttgctcctgccgagaaagtatccatcatggctgatgcaatgc 
               
               
                   
                 ggcggctgcatacgcttgatccggctacctgcccattcgaccaccaagcg 
               
               
                   
                 aaacatcgcatcgagcgagcacgtactcggatggaagccggtcttgtcga 
               
               
                   
                 tcaggatgatctggacgaagagcatcaggggctcgcgccagccgaactgt 
               
               
                   
                 tcgccaggctcaaggcgcgcatgcccgacggcgaggatctcgtcgtgacc 
               
               
                   
                 catggcgatgcctgcttgccgaatatcatggtggaaaatggccgcttttc 
               
               
                   
                 tggattcatcgactgtggccggctgggtgtggcggaccgctatcaggaca 
               
               
                   
                 tagcgttggctacccgtgatattgctgaagagcttggcggcgaatgggct 
               
               
                   
                 gaccgcttcctcgtgctttacggtatcgccgctcccgattcgcagcgcat 
               
               
                   
                 cgccttctatcgccttcttgacgagttcttctgagcgggactctggggtt 
               
               
                   
                 cgaaatgaccgaccaagcgacgcccaacctgccatcacgagatttcgatt 
               
               
                   
                 ccaccgccgccttctatgaaaggttgggcttcggaatcgttttccgggac 
               
               
                   
                 gccggctggatgatcctccagcgcggggatctcatgctggagttcttcgc 
               
               
                   
                 ccaccccagcttcaaaagcgctctgaagttcctatactttctagagaata 
               
               
                   
                 ggaacttcggaataggaactaaggaggatattcatatatgcattaatgtc 
               
               
                   
                 taacaattcgttcaagccgacgccgcttcgcggcgcggcttaactcaagc 
               
               
                   
                 gttagatgcactaagcacataattgctcacagccaaactatcaggtcaag 
               
               
                   
                 tctgcttttattatttttaagcgtgcataataagccctacacaaattggg 
               
               
                   
                 agatatatcatgaaaggctggctttttcttgttatcgcaatagttgCGAT 
               
               
                   
                 CGTTATGACAACTTGACGGCTACATCATTCACTTTTTCTTCACAACCGGC 
               
               
                   
                 ACGGAACTCGCTCGGGCTGGCCCCGGTGCATTTTTTAAATACCCGCGAGA 
               
               
                   
                 AGTAGAGTTGATCGTCAAAACCAACATTGCGACCGACGGTGGCGATAGGC 
               
               
                   
                 ATCCGGGTGGTGCTCAAAAGCAGCTTCGCCTGGCTGATACGTTGGTCCTC 
               
               
                   
                 GCGCCAGCTTAAGACGCTAATCCCTAACTGCTGGCGGAAAAGATGTGACA 
               
               
                   
                 GACGCGACGGCGACAAGCAAACATGCTGTGCGACGCTGGCGATATCAAAA 
               
               
                   
                 TTGCTGTCTGCCAGGTGATCGCTGATGTACTGACAAGCCTCGCGTACCCG 
               
               
                   
                 ATTATCCATCGGTGGATGGAGCGACTCGTTAATCGCTTCCATGCGCCGCA 
               
               
                   
                 GTAACAATTGCTCAAGCAGATTTATCGCCAGCAGCTCCGAATAGCGCCCT 
               
               
                   
                 TCCCCTTGCCCGGCGTTAATGATTTGCCCAAACAGGTCGCTGAAATGCGG 
               
               
                   
                 CTGGTGCGCTTCATCCGGGCGAAAGAACCCCGTATTGGCAAATATTGACG 
               
               
                   
                 GCCAGTTAAGCCATTCATGCCAGTAGGCGCGCGGACGAAAGTAAACCCAC 
               
               
                   
                 TGGTGATACCATTCGCGAGCCTCCGGATGACGACCGTAGTGATGAATCTC 
               
               
                   
                 TCCTGGCGGGAACAGCAAAATAACACCCGGTCGGCAAACAAATTCTCGTC 
               
               
                   
                 CCTGATTTTTCACCACCCCCTGACCGCGAATGGTGAGATTGAGAATATAA 
               
               
                   
                 CCTTTCATTCCCAGCGGTCGGTCGATAAAAAAATCGAGATAACCGTTGGC 
               
               
                   
                 CTCAATCGGCGTTAAACCCGCCACCAGATGGGCATTAAACGAGTATCCCG 
               
               
                   
                 GCAGCAGGGGATCATTTTGCGCTTCAGCCATACTTTTCATACTCCCGCCA 
               
               
                   
                 TTCAGAGAAGAAACCAATTGTCCATATTGCATCAGACATTGCCGTCACTG 
               
               
                   
                 CGTCTTTTACTGGCTCTTCTCGCTAACCAAACCGGTAACCCCGCTTATTA 
               
               
                   
                 AAAGCATTCTGTAACAAAGCGGGACCAAAGCCATGACAAAAACGCGTAAC 
               
               
                   
                 AAAAGTGTCTATAATCACGGCAGAAAAGTCCACATTGATTATTTGCACGG 
               
               
                   
                 CGTCACACTTTGCTATGCCATAGCATTTTTATCCATAAGATTAGCGGATC 
               
               
                   
                 CTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACCCGTTT 
               
               
                   
                 TTTTGGGAATTCGAGCTCGGAGTTAACAAAAGATGTTAGATGCAAACAAA 
               
               
                   
                 TTACAGCAGGCAGTGGATCAGGCTTACACCCAATTTCACTCACTTAACGG 
               
               
                   
                 CGGACAAAATGCCGATTACATTCCCTTTCTGGCGAATGTACCAGGTCAAC 
               
               
                   
                 TGGCGGCAGTGGCTATCGTGACNTGCGATGGCAACGTCTATAGTGCGGGT 
               
               
                   
                 GACAGTGATTACCGCTTTGCACTGGAATCCATCTCTAAAGTCTGTACGTT 
               
               
                   
                 AGCCCTTGCGTTAGAAGATGTCGGCCCGCAGGCGGTACAGGACAAAATTG 
               
               
                   
                 GCGCTGACCCGACCGGATTGCCCTTTAACTCAGTTATCGCCTTAGAGTTG 
               
               
                   
                 CATGGCGGCAAACCGCTGTCGCCACTGGTAAATGCTGGCGCTATTGCCAC 
               
               
                   
                 CACCAGCCTGATTAACGCTGAAAATGTTGAACAACGCTGGCAGCGAATTT 
               
               
                   
                 TACATATCCAACAGCAACTGGCTGGTGAGCAGGTAGCGCTCTCTGACGAA 
               
               
                   
                 GTCAACCAGTCGGAACAAACAACCAACTTCCATAACCGGGCCATTGCCTG 
               
               
                   
                 GCTGCTGTACTCCGCCGGATATCTCTATTGTGATGCAATGGAAGCCTGTG 
               
               
                   
                 ACGTGTACACCCGTCAGTGCTCTACGCTCATCAATACTGTTGAACTGGCA 
               
               
                   
                 ACGCTTGGCGCGACGCTGGCGGCGGGTGGTGTGAATCCGTTGACGCATAA 
               
               
                   
                 ACGCGTTCTTCAGGCCGACAACGTGCCGTACATTCTGGCCGAAATGATGA 
               
               
                   
                 TGGAAGGGCTGTATGGTCGCTCCGGTGACTGGGCGTATCGCGTTGGTTTA 
               
               
                   
                 CCGGGCAAAAGCGGTGTAGGTGGCGGTATTCTGGCGGTCGTCCCTGGCGT 
               
               
                   
                 GATGGGAATTGCCGCGTTCTCACCACCGCTGGACGAAGAAGGCAACAGTG 
               
               
                   
                 TTCGCGGTCAAAAAATGGTGGCATCGGTCGCTAAGCAACTCGGCTATAAC 
               
               
                   
                 GTGTTTAAGGGCTGACCATGGAGTGTTTTAATGCGATCCAATCATTTTAA 
               
               
                   
                 GGAGTTTAAAATGGATAAGAAGCAAGTAACGGATTTAAGGTCGGAACTAC 
               
               
                   
                 TCGATTCACGTTTTGGTGCGAAGTCTATTTCCACTATCGCAGAATCAAAA 
               
               
                   
                 CGTTTTCCGCTGCACGAAATGCGCGACGATGTCGCATTTCAGATTATCAA 
               
               
                   
                 TGATGAATTATATCTTGATGGCAACGCTCGTCAGAACCTGGCCACTTTCT 
               
               
                   
                 GCCAGACCTGGGACGACGAAAACGTCCACAAGTTGATGGATTTATCCATT 
               
               
                   
                 AACAAAAACTGGATCGACAAAGAAGAATATCCGCAATCCGCAGCCATCGA 
               
               
                   
                 CCTGCGTTGCGTAAACATGGTTGCCGATCTGTGGCATGCGCCTGCGCCGA 
               
               
                   
                 AAAATGGTCAGGCCGTTGGCACCAACACCATTGGTTCTTCCGAGGCCTGT 
               
               
                   
                 ATGCTCGGCGGGATGGCGATGAAATGGCGTTGGCGCAAGCGTATGGAAGC 
               
               
                   
                 TGCAGGCAAACCAACGGATAAACCAAACCTGGTGTGCGGTCCGGTGCAAA 
               
               
                   
                 TCTGCTGGCATAAATTCGCCCGCTACTGGGATGTGGAGCTGCGTGAGATC 
               
               
                   
                 CCTATGCGCCCCGGTCAGTTGTTTATGGACCCGAAACGCATGATTGAAGC 
               
               
                   
                 CTGCGACGAAAATACCATCGGCGTGGTGCCGACTTTCGGCGTGACCTACA 
               
               
                   
                 CCGGTAACTATGAGTTCCCGCAACCGCTGCACGATGCACTGGATAAATTC 
               
               
                   
                 CAGGCCGACACCGGTATCGACATCGACATGCACATCGACGCCGCCAGCGG 
               
               
                   
                 TGGCTTTCTGGCACCGTTCGTCGCCCCGGATATCGTCTGGGACTTCCGCC 
               
               
                   
                 TGCCGCGTGTGAAATCGATCAGTGCTTCAGGCCATAAATTCGGTCTGGCT 
               
               
                   
                 CCGCTGGGCTGCGGCTGGGTTATCTGGCGTGACGAAGAAGCGCTGCCGCA 
               
               
                   
                 GGAACTGGTGTTCAACGTTGACTACCTCGGTGGTCAGATTGGTACTTTTG 
               
               
                   
                 CCATCAACTTCTCCCGCCCGGCGGGTCAGGTGATTGCACAGTACTATGAA 
               
               
                   
                 TTCCTGCGCCTTGGTCGTGAAGGCTATACCAAAGTACAGAATGCTTCCTA 
               
               
                   
                 CCAGGTCGCTGCCTATCTGGCGGATGAGATCGCCAAACTGGGGCCGTATG 
               
               
                   
                 AGTTCATCTGTACCGGTCGCCCGGACGAAGGCATCCCGGCGGTTTGCTTC 
               
               
                   
                 AAACTGAAAGATGGTGAAGATCCGGGATACACCCTGTACGACCTCTCTGA 
               
               
                   
                 ACGTCTGCGTCTGCGCGGCTGGCAGGTTCCGGCCTTCACTCTCGGCGGTG 
               
               
                   
                 AAGCCACCGACATCGTGGTGATGCGCATTATGTGTCGTCGCGGCTTCGAA 
               
               
                   
                 ATGGACTTTGCTGAACTGTTGCTGGAAGACTACAAAGCCTCCCTGAAATA 
               
               
                   
                 TCTCAGCGATCACCCGAAACTGCAGGGTATTGCCCAGCAGAACAGCTTTA 
               
               
                   
                 AACATACCTGATAACCGTTTTGGAGTGATAATATGGCTACATCAGTACAG 
               
               
                   
                 ACAGGTAAAGCTAAGCAGCTCACATTACTCGGATTCTTTGCCATAACGGC 
               
               
                   
                 ATCGATGGTAATGGCTGTTTATGAATACCCTACCTTCGCAACATCGGGCT 
               
               
                   
                 TTTCATTAGTCTTCTTCCTGCTATTAGGCGGGATTTTATGGTTTATTCCC 
               
               
                   
                 GTGGGACTTTGTGCTGCGGAAATGGCCACCGTCGACGGCTGGGAAGAAGG 
               
               
                   
                 TGGTGTCTTCGCCTGGGTATCAAATACTCTGGGTCCGAGATGGGGATTTG 
               
               
                   
                 CAGCGATCTCATTTGGCTATCTGCAAATCGCCATTGGTTTTATTCCGATG 
               
               
                   
                 CTCTATTTCGTGTTAGGGGCACTCTCCTACATCCTGAAATGGCCAGCGCT 
               
               
                   
                 GAATGAAGACCCCATTACCAAAACTATTGCAGCACTCATCATTCTTTGGG 
               
               
                   
                 CGCTGGCATTAACGCAGTTTGGTGGCACGAAATACACGGCGCGAATTGCT 
               
               
                   
                 AAAGTTGGCTTCTTCGCCGGTATCCTGTTACCTGCATTTATTTTGATCGC 
               
               
                   
                 ATTAGCGGCTATTTACCTGCACTCCGGTGCCCCCGTTGCTATCGAAATGG 
               
               
                   
                 ATTCGAAGACCTTCTTCCCTGACTTCTCTAAAGTGGGCACACTGGTTGTA 
               
               
                   
                 TTTGTTGCCTTCATTTTGAGTTATATGGGCGTAGAAGCTTCCGCAACTCA 
               
               
                   
                 CGTCAATGAAATGAGCAACCCCGGGCGCGACTATCCACTGGCTATGTTAC 
               
               
                   
                 TGCTGATGGTGGCGGCAATCTGCTTAAGCTCTGTTGGCGGTTTGTCTATT 
               
               
                   
                 GCGATGGTCATTCCGGGTAATGAAATCAACCTCTCCGCAGGGGTAATGCA 
               
               
                   
                 AACCTTTACCGTTCTGATGTCCCATGTGGCACCGGAAATTGAGTGGACGG 
               
               
                   
                 TTCGCGTGATCTCCGCACTGCTGTTGCTGGGTGTTCTGGCGGAAATCGCC 
               
               
                   
                 TCCTGGATTGTTGGTCCTTCTCGCGGGATGTATGTCACAGCGCAGAAAAA 
               
               
                   
                 CCTGCTGCCAGCGGCATTCGCTAAAATGAACAAAAATGGCGTACCGGTAA 
               
               
                   
                 CGCTGGTCATTTCGCAGCTGGTGATTACTTCTATCGCGTTGATCATCCTC 
               
               
                   
                 ACCAATACCGGTGGCGGTAACAACATGTCCTTCCTGATCGCACTGGCGCT 
               
               
                   
                 GACGGTGGTGATTTATCTGTGTGCTTATTTCATGCTGTTTATTGGCTACA 
               
               
                   
                 TTGTGTTGGTTCTTAAACATCCTGACTTAAAACGCACATTTAATATCCCT 
               
               
                   
                 GGTGGTAAAGGGGTGAAACTGGTCGTGGCAATTGTCGGTCTGCTGACTTC 
               
               
                   
                 AATTATGGCGTTTATTGTTTCCTTCCTGCCGCCGGATAACATCCAGGGTG 
               
               
                   
                 ATTCTACCGATATGTATGTTGAATTACTGGTTGTTAGTTTCCTGGTGGTA 
               
               
                   
                 CTTGCCCTGCCCTTTATTCTCTATGCTGTTCATGATCGTAAAGGCAAAGC 
               
               
                   
                 GAATACCGGCGTCACTCTGGAGCCAATCAACAGTCAGAACGCACCAAAAG 
               
               
                   
                 GTCACTTCTTCCTGCACCCGCGTGCACGTTCACCACACTATATTGTGATG 
               
               
                   
                 AATGACAAGAAACACTAACGATCGgcgaagtaatcgcaacatccgcatta 
               
               
                   
                 aaatctagcgagggctttactgtcgaccaaaaaaatattgacaacataaa 
               
               
                   
                 aaactttgtgttatacttgtaacgGactctagagggtattaataatgaaa 
               
               
                   
                 ggagaggacatATGAGCGATAACGGCCCGCAGAACCAGCGCAACGCGCCG 
               
               
                   
                 CGCATTACCTTTGGCGGCCCGAGCGATAGCACCGGCAGCAACCAGAACGG 
               
               
                   
                 CGAACGCAGCGGCGCGCGCAGCAAACAGCGCCGCCCGCAGGGCCTGCCGA 
               
               
                   
                 ACAACACCGCGAGCTGGTTTACCGCGCTGACCCAGCATGGCAAAGAAGAT 
               
               
                   
                 CTGAAATTTCCGCGCGGCCAGGGCGTGCCGATTAACACCAACAGCAGCCC 
               
               
                   
                 GGATGATCAGATTGGCTATTATCGCCGCGCGACCCGCCGCATTCGCGGCG 
               
               
                   
                 GCGATGGCAAAATGAAAGATCTGAGCCCGCGCTGGTATTTTTATTATCTG 
               
               
                   
                 GGCACCGGCCCGGAAGCGGGCCTGCCGTATGGCGCGAACAAAGATGGCAT 
               
               
                   
                 TATTTGGGTGGCGACCGAAGGCGCGCTGAACACCCCGAAAGATCATATTG 
               
               
                   
                 GCACCCGCAACCCGGCGAACAACGCGGCGATTGTGCTGCAGCTGCCGCAG 
               
               
                   
                 GGCACCACCCTGCCGAAAGGCTTTTATGCGGAAGGCAGCCGCGGCGGCAG 
               
               
                   
                 CCAGGCGAGCAGCCGCAGCAGCAGCCGCAGCCGCAACAGCAGCCGCAACA 
               
               
                   
                 GCACCCCGGGCAGCAGCCGCGGCACCAGCCCGGCGCGCATGGCGGGCAAC 
               
               
                   
                 GGCGGCGATGCGGCGCTGGCGCTGCTGCTGCTGGATCGCCTGAACCAGCT 
               
               
                   
                 GGAAAGCAAAATGAGCGGCAAAGGCCAGCAGCAGCAGGGCCAGACCGTGA 
               
               
                   
                 CCAAAAAAAGCGCGGCGGAAGCGAGCAAAAAACCGCGCCAGAAACGCACC 
               
               
                   
                 GCGACCAAAGCGTATAACGTGACCCAGGCGTTTGGCCGCCGCGGCCCGGA 
               
               
                   
                 ACAGACCCAGGGCAACTTTGGCGATCAGGAACTGATTCGCCAGGGCACCG 
               
               
                   
                 ATTATAAACATTGGCCGCAGATTGCGCAGTTTGCGCCGAGCGCGAGCGCG 
               
               
                   
                 TTTTTTGGCATGAGCCGCATTGGCATGGAAGTGACCCCGAGCGGCACCTG 
               
               
                   
                 GCTGACCTATACCGGCGCGATTAAACTGGATGATAAAGATCCGAACTTTA 
               
               
                   
                 AAGATCAGGTGATTCTGCTGAACAAACATATTGATGCGTATAAAACCTTT 
               
               
                   
                 CCGCCGACCGAACCGAAAAAAGATAAAAAAAAAAAAGCGGATGAAACCCA 
               
               
                   
                 GGCGCTGCCGCAGCGCCAGAAAAAACAGCAGACCGTGACCCTGCTGCCGG 
               
               
                   
                 CGGCGGATCTGGATGATTTTAGCAAACAGCTGCAGCAGAGCATGAGCAGC 
               
               
                   
                 GCGGATAGCACCCAGGCGCCTAGGCTTAAGATGGCGGATAGCAACGGCAC 
               
               
                   
                 CATTACCGTGGAAGAACTGAAAAAACTGCTGGAACAGTGGAACCTGGTGA 
               
               
                   
                 TTGGCTTTCTGTTTCTGACCTGGATTTGCCTGCTGCAGTTTGCGTATGCG 
               
               
                   
                 AACCGCAACCGCTTTCTGTATATTATTAAACTGATTTTTCTGTGGCTGCT 
               
               
                   
                 GTGGCCGGTGACCCTGGCGTGCTTTGTGCTGGCGGCGGTGTATCGCATTA 
               
               
                   
                 ACTGGATTACCGGCGGCATTGCGATTGCGATGGCGTGCCTGGTGGGCCTG 
               
               
                   
                 ATGTGGCTGAGCTATTTTATTGCGAGCTTTCGCCTGTTTGCGCGCACCCG 
               
               
                   
                 CAGCATGTGGAGCTTTAACCCGGAAACCAACATTCTGCTGAACGTGCCGC 
               
               
                   
                 TGCATGGCACCATTCTGACCCGCCCGCTGCTGGAAAGCGAACTGGTGATT 
               
               
                   
                 GGCGCGGTGATTCTGCGCGGCCATCTGCGCATTGCGGGCCATCATCTGGG 
               
               
                   
                 CCGCTGCGATATTAAAGATCTGCCGAAAGAAATTACCGTGGCGACCAGCC 
               
               
                   
                 GCACCCTGAGCTATTATAAACTGGGCGCGAGCCAGCGCGTGGCGGGCGAT 
               
               
                   
                 AGCGGCTTTGCGGCGTATAGCCGCTATCGCATTGGCAACTATAAACTGAA 
               
               
                   
                 CACCGATCATAGCAGCAGCAGCGATAACATTGCGCTGCTGGTGCAGCTTA 
               
               
                   
                 AGGAGCTCATGTTTGTGTTTCTGGTGCTGCTGCCGCTGGTGAGCAGCCAG 
               
               
                   
                 TGCGTGAACCTGACCACCCGCACCCAGCTGCCGCCGGCGTATACCAACAG 
               
               
                   
                 CTTTACCCGCGGCGTGTATTATCCGGATAAAGTGTTTCGCAGCAGCGTGC 
               
               
                   
                 TGCATAGCACCCAGGATCTGTTTCTGCCGTTTTTTAGCAACGTGACCTGG 
               
               
                   
                 TTTCATGCGATTCATGTGAGCGGCACCAACGGCACCAAACGCTTTGATAA 
               
               
                   
                 CCCGGTGCTGCCGTTTAACGATGGCGTGTATTTTGCGAGCACCGAAAAAA 
               
               
                   
                 GCAACATTATTCGCGGCTGGATTTTTGGCACCACCCTGGATAGCAAAACC 
               
               
                   
                 CAGAGCCTGCTGATTGTGAACAACGCGACCAACGTGGTGATTAAAGTGTG 
               
               
                   
                 CGAATTTCAGTTTTGCAACGATCCGTTTCTGGGCGTGTATTATCATAAAA 
               
               
                   
                 ACAACAAAAGCTGGATGGAAAGCGAATTTCGCGTGTATAGCAGCGCGAAC 
               
               
                   
                 AACTGCACCTTTGAATATGTGAGCCAGCCGTTTCTGATGGATCTGGAAGG 
               
               
                   
                 CAAACAGGGCAACTTTAAAAACCTGCGCGAATTTGTGTTTAAAAACATTG 
               
               
                   
                 ATGGCTATTTTAAAATTTATAGCAAACATACCCCGATTAACCTGGTGCGC 
               
               
                   
                 GATCTGCCGCAGGGCTTTAGCGCGCTGGAACCGCTGGTGGATCTGCCGAT 
               
               
                   
                 TGGCATTAACATTACCCGCTTTCAGACCCTGCTGGCGCTGCATCGCAGCT 
               
               
                   
                 ATCTGACCCCGGGCGATAGCAGCAGCGGCTGGACCGCGGGCGCGGCGGCG 
               
               
                   
                 TATTATGTGGGCTATCTGCAGCCGCGCACCTTTCTGCTGAAATATAACGA 
               
               
                   
                 AAACGGCACCATTACCGATGCGGTGGATTGCGCGCTGGATCCGCTGAGCG 
               
               
                   
                 AAACCAAATGCACCCTGAAAAGCTTTACCGTGGAAAAAGGCATTTATCAG 
               
               
                   
                 ACCAGCAACTTTCGCGTGCAGCCGACCGAAAGCATTGTGCGCTTTCCGAA 
               
               
                   
                 CATTACCAACCTGTGCCCGTTTGGCGAAGTGTTTAACGCGACCCGCTTTG 
               
               
                   
                 CGAGCGTGTATGCGTGGAACCGCAAACGCATTAGCAACTGCGTGGCGGAT 
               
               
                   
                 TATAGCGTGCTGTATAACAGCGCGAGCTTTAGCACCTTTAAATGCTATGG 
               
               
                   
                 CGTGAGCCCGACCAAACTGAACGATCTGTGCTTTACCAACGTGTATGCGG 
               
               
                   
                 ATAGCTTTGTGATTCGCGGCGATGAAGTGCGCCAGATTGCGCCGGGCCAG 
               
               
                   
                 ACCGGCAAAATTGCGGATTATAACTATAAACTGCCGGATGATTTTACCGG 
               
               
                   
                 CTGCGTGATTGCGTGGAACAGCAACAACCTGGATAGCAAAGTGGGCGGCA 
               
               
                   
                 ACTATAACTATCTGTATCGCCTGTTTCGCAAAAGCAACCTGAAACCGTTT 
               
               
                   
                 GAACGCGATATTAGCACCGAAATTTATCAGGCGGGCAGCACCCCGTGCAA 
               
               
                   
                 CGGCGTGGAAGGCTTTAACTGCTATTTTCCGCTGCAGAGCTATGGCTTTC 
               
               
                   
                 AGCCGACCAACGGCGTGGGCTATCAGCCGTATCGCGTGGTGGTGCTGAGC 
               
               
                   
                 TTTGAACTGCTGCATGCGCCGGCGACCGTGTGCGGCCCGAAAAAAAGCAC 
               
               
                   
                 CAACCTGGTGAAAAACAAATGCGTGAACTTTAACTTTAACGGCCTGACCG 
               
               
                   
                 GCACCGGCGTGCTGACCGAAAGCAACAAAAAATTTCTGCCGTTTCAGCAG 
               
               
                   
                 TTTGGCCGCGATATTGCGGATACCACCGATGCGGTGCGCGATCCGCAGAC 
               
               
                   
                 CCTGGAAATTCTGGATATTACCCCGTGCAGCTTTGGCGGCGTGAGCGTGA 
               
               
                   
                 TTACCCCGGGCACCAACACCAGCAACCAGGTGGCGGTGCTGTATCAGGGC 
               
               
                   
                 GTGAACTGCACCGAAGTGCCGGTGGCGATTCATGCGGATCAGCTGACCCC 
               
               
                   
                 GACCTGGCGCGTGTATAGCACCGGCAGCAACGTGTTTCAGACCCGCGCGG 
               
               
                   
                 GCTGCCTGATTGGCGCGGAACATGTGAACAACAGCTATGAATGCGATATT 
               
               
                   
                 CCGATTGGCGCGGGCATTTGCGCGAGCTATCAGACCCAGACCAACAGCCC 
               
               
                   
                 GGGCAGCGCGAGCAGCGTGGCGAGCCAGAGCATTATTGCGTATACCATGA 
               
               
                   
                 GCCTGGGCGCGGAAAACAGCGTGGCGTATAGCAACAACAGCATTGCGATT 
               
               
                   
                 CCGACCAACTTTACCATTAGCGTGACCACCGAAATTCTGCCGGTGAGCAT 
               
               
                   
                 GACCAAAACCAGCGTGGATTGCACCATGTATATTTGCGGCGATAGCACCG 
               
               
                   
                 AATGCAGCAACCTGCTGCTGCAGTATGGCAGCTTTTGCACCCAGCTGAAC 
               
               
                   
                 CGCGCGCTGACCGGCATTGCGGTGGAACAGGATAAAAACACCCAGGAAGT 
               
               
                   
                 GTTTGCGCAGGTGAAACAGATTTATAAAACCCCGCCGATTAAAGATTTTG 
               
               
                   
                 GCGGCTTTAACTTTAGCCAGATTCTGCCGGATCCGAGCAAACCGAGCAAA 
               
               
                   
                 CGCAGCTTTATTGAAGATCTGCTGTTTAACAAAGTGACCCTGGCGGATGC 
               
               
                   
                 GGGCTTTATTAAACAGTATGGCGATTGCCTGGGCGATATTGCGGCGCGCG 
               
               
                   
                 ATCTGATTTGCGCGCAGAAATTTAACGGCCTGACCGTGCTGCCGCCGCTG 
               
               
                   
                 CTGACCGATGAAATGATTGCGCAGTATACCAGCGCGCTGCTGGCGGGCAC 
               
               
                   
                 CATTACCAGCGGCTGGACCTTTGGCGCGGGCGCGGCGCTGCAGATTCCGT 
               
               
                   
                 TTGCGATGCAGATGGCGTATCGCTTTAACGGCATTGGCGTGACCCAGAAC 
               
               
                   
                 GTGCTGTATGAAAACCAGAAACTGATTGCGAACCAGTTTAACAGCGCGAT 
               
               
                   
                 TGGCAAAATTCAGGATAGCCTGAGCAGCACCGCGAGCGCGCTGGGCAAAC 
               
               
                   
                 TGCAGGATGTGGTGAACCAGAACGCGCAGGCGCTGAACACCCTGGTGAAA 
               
               
                   
                 CAGCTGAGCAGCAACTTTGGCGCGATTAGCAGCGTGCTGAACGATATTCT 
               
               
                   
                 GAGCCGCCTGGATCCGCCGGAAGCGGAAGTGCAGATTGATCGCCTGATTA 
               
               
                   
                 CCGGCCGCCTGCAGAGCCTGCAGACCTATGTGACCCAGCAGCTGATTCGC 
               
               
                   
                 GCGGCGGAAATTCGCGCGAGCGCGAACCTGGCGGCGACCAAAATGAGCGA 
               
               
                   
                 ATGCGTGCTGGGCCAGAGCAAACGCGTGGATTTTTGCGGCAAAGGCTATC 
               
               
                   
                 ATCTGATGAGCTTTCCGCAGAGCGCGCCGCATGGCGTGGTGTTTCTGCAT 
               
               
                   
                 GTGACCTATGTGCCGGCGCAGGAAAAAAACTTTACCACCGCGCCGGCGAT 
               
               
                   
                 TTGCCATGATGGCAAAGCGCATTTTCCGCGCGAAGGCGTGTTTGTGAGCA 
               
               
                   
                 ACGGCACCCATTGGTTTGTGACCCAGCGCAACTTTTATGAACCGCAGATT 
               
               
                   
                 ATTACCACCGATAACACCTTTGTGAGCGGCAACTGCGATGTGGTGATTGG 
               
               
                   
                 CATTGTGAACAACACCGTGTATGATCCGCTGCAGCCGGAACTGGATAGCT 
               
               
                   
                 TTAAAGAAGAACTGGATAAATATTTTAAAAACCATACCAGCCCGGATGTG 
               
               
                   
                 GATCTGGGCGATATTAGCGGCATTAACGCGAGCGTGGTGAACATTCAGAA 
               
               
                   
                 AGAAATTGATCGCCTGAACGAAGTGGCGAAAAACCTGAACGAAAGCCTGA 
               
               
                   
                 TTGATCTGCAGGAACTGGGCAAATATGAACAGGGCTATATTCCGGAAGCG 
               
               
                   
                 CCGCGCGATGGCCAGGCGTATGTGCGCAAAGATGGCGAATGGGTGCTGCT 
               
               
                   
                 GAGCACCTTTCTGGCATTAGCCTATGGAAGTCAGGGTAATCTTAATCCAT 
               
               
                   
                 TAATTAATGAAATCAGCAAAATCATTTCAGCTGCAGGTAATTTTGATGTT 
               
               
                   
                 AAAGAGGAAAGAGCGGCCGCGTCTTTATTGCAGTTGTCCGGTAATGCCAG 
               
               
                   
                 TGATTTTTCATATGGACGGAACTCAATAACTTTGACAGCATCAGCATAAG 
               
               
                   
                 AGCTCtttattaatttaaataatagcaatcttactgggctgtgccacata 
               
               
                   
                 agattgctatttttttggagtcataatggattcttgtcataaaattgatt 
               
               
                   
                 atgggttatacgccctggagattttagcccaataccataacgtctctgtt 
               
               
                   
                 aacccggaagaaattaaacatagatttgacacagacgggactggtctggg 
               
               
                   
                 attaacgtcatggttgcttgctgcgaaatctttagaactaaaggtaaaac 
               
               
                   
                 aggtaaaaaaaacaattgaccgattaaactttatttctctgcccgcatta 
               
               
                   
                 gtctggagagaggatggatgtcattttattctgactaaagtcagtaaaga 
               
               
                   
                 agcaaacagatatcttatttttgatctggagcagcgaaatccccgtgttc 
               
               
                   
                 tcgaacagtctgagtttgaggcgttatatcaggggcatattattcttatt 
               
               
                   
                 gcttcccgttcttctgttaccgggaaactggcaaaatttgactttacctg 
               
               
                   
                 gtttattcctgccattataaaatacaggaaaatatttattgaaacccttg 
               
               
                   
                 ttgtatctgtttttttacaattatttgcattaataacccccctttttttt 
               
               
                   
                 caggtggttatggacaaagtattagtacacagggggttttcaacccttaa 
               
               
                   
                 tgttattactgtcgcattatctgttgtggtggtgtttgagattatactca 
               
               
                   
                 gcggtttaagaacttacatttttgcacatagtacaagtcggattgatgtt 
               
               
                   
                 gagttgggtgccaaactcttccggcatttactggcgctaccgatctctta 
               
               
                   
                 ttttgagagtcgtcgtgttggtgatactgttgccagggtaagagaattag 
               
               
                   
                 accagatccgtaatttcctgacaggacaggcattaacatctgttctggac 
               
               
                   
                 ttattattttcattcatattttttgcggtaatgtggtattacagcccaaa 
               
               
                   
                 gcttactctggtgatcttattttcgctgccctgttatgctgcatggtctg 
               
               
                   
                 tttttattagccccattttgcgacgtcgccttgatgataagttttcacgg 
               
               
                   
                 aatgcggataatcaatctttcctggtggaatcagtcacggcgattaacac 
               
               
                   
                 tataaaagctatggcagtctcacctcagatgacgaacatatgggacaaac 
               
               
                   
                 aattggcaggatatgttgctgcaggctttaaagtgacagtattagccacc 
               
               
                   
                 attggtcaacaaggaatacagttaatacaaaagactgttatgatcatcaa 
               
               
                   
                 cctgtggttgggtgcacacctggttatttccggggatttaagtattggtc 
               
               
                   
                 agttaattgcttttaatatgcttgcaggtcagattgttgcaccggttatt 
               
               
                   
                 cgccttgcacaaatctggcaggatttccagcaggttggtatatcagttac 
               
               
                   
                 ccgccttggtgatgtgcttaactctccaactgaaagttatcatgggaaac 
               
               
                   
                 tggcattaccggaaattaatggtgatatcacttttcgtaatatccggttt 
               
               
                   
                 cgctataagcctgactctccggttattttagataatatcaatctcagtat 
               
               
                   
                 taagcagggggaggttattggtattgtcggacgttctggttcaggaaaaa 
               
               
                   
                 gcacattaactaaattaattcaacgtttttatattcctgaaaatggccag 
               
               
                   
                 gtcttaattgatggacatgatcttgcgttggccgatcctaactggttacg 
               
               
                   
                 tcgtcaggtgggggttgtgttgcaggacaatgtgctgcttaatcgcagta 
               
               
                   
                 ttattgataatatttcactggctaatcctggcatgtccgtcgaaaaagtt 
               
               
                   
                 atttatgcagcgaaattagcaggtgctcatgattttatttctgaattgcg 
               
               
                   
                 tgaggggtataacaccattgtcggggaacagggggcaggattatccggag 
               
               
                   
                 gtcaacgtcaacgcatcgcaattgcaagggcgctggtgaacaaccctaaa 
               
               
                   
                 atactcatctttgatgaagcaaccagtgctctggattatgagtcggagca 
               
               
                   
                 tgtcatcatgcgcaatatgcacaaaatatgtaagggcagaacggttataa 
               
               
                   
                 tcattgctcatcgtctgtctacagtaaaaaatgcagaccgcattattgtc 
               
               
                   
                 atggaaaaagggaaaattgttgaacagggtaaacataaggagctgctttc 
               
               
                   
                 tgaaccggaaagtttatacagttacttatatcagttacagtcagactaac 
               
               
                   
                 agaaagaacagaagaatatgaaaacatggttaatggggttcagcgagttc 
               
               
                   
                 ctgttgcgctataaacttgtctggagtgaaacatggaaaatccggaagca 
               
               
                   
                 gttagatactccggtacgtgaaaaggacgaaaatgaattcttacccgctc 
               
               
                   
                 atctggaattaattgaaacgccggtatccagacggccgcgtctggttgct 
               
               
                   
                 tattttattatggggtttctggttattgctttcattttatctgttttagg 
               
               
                   
                 ccaggtggaaattgttgccactgcaaatgggaaattaacactcagtgggc 
               
               
                   
                 gtagcaaagaaattaaacctattgaaaactcgatagttaaagaaattatc 
               
               
                   
                 gtaaaagaaggagagtcagtccggaaaggggatgtgttattaaagcttac 
               
               
                   
                 agcgctgggagctgaagctgatacgttaaaaacgcagtcatcactgttac 
               
               
                   
                 aggccaggctggaacaaattcggtatcaaattctgagccggtcaattgaa 
               
               
                   
                 ttaaataaacttcctgaactgaaacttcctgatgagccttattttcagaa 
               
               
                   
                 tgtatctgaagaggaagtactgcgtttaacttctttgataaaagaacagt 
               
               
                   
                 tttccacatggcaaaatcagaagtatcaaaaagaactgaatctggataag 
               
               
                   
                 aaaagagcagagcgattaacaatacttgcccgtataaaccgttatgaaaa 
               
               
                   
                 tgtatcgagggttgaaaaaagccgtctggatgatttcaggagcctgttgc 
               
               
                   
                 ataaacaggcaattgcaaaacatgctgtacttgagcaggagaataaatat 
               
               
                   
                 gttgaggcagcaaatgaattacgggtttataaatcgcaactggagcaaat 
               
               
                   
                 tgagagtgagatattgtctgcaaaagaagaatatcagcttgtcacgcagc 
               
               
                   
                 tttttaaaaatgaaattttagacaagctaagacaaacaacagacagcatt 
               
               
                   
                 gagttattaactctggagttagagaaaaatgaagagcgtcaacaggcttc 
               
               
                   
                 agtaatcagggcccctgtttcgggaaaagttcagcaactgaaggttcata 
               
               
                   
                 ctgaaggtggggttgttacaacagcggaaacactgatggtcatcgttccg 
               
               
                   
                 gaagatgacacgctggaggttactgctctggtacaaaataaagatattgg 
               
               
                   
                 ttttattaacgtcgggcagaatgccatcattaaagtggaggcttttcctt 
               
               
                   
                 acacccgatatggttatctggtgggtaaggtaaaaaatataaatttagat 
               
               
                   
                 gcaatagaagaccagaaactgggactcgtttttaatgtcattgtttctgt 
               
               
                   
                 tgaagagaatgatttgtcaaccgggaataagcacattccattaagctcgg 
               
               
                   
                 gtatggctgtcactgcagaaataaagactggaatgcgaagcgtaatcagt 
               
               
                   
                 tatcttcttagtcccctggaagaatctgtaacagaaagtttacatgagcg 
               
               
                   
                 ttaagtctcagagcagcggtatccggctcatatcttctcctgtcagatcc 
               
               
                   
                 aagggcgaattccagcacactggcggccgttactagtggatccaccggtc 
               
               
                   
                 tcgaggggcgcgccggtaccgaattctaattaatgggcatcatttttcag 
               
               
                   
                 ctatttcatttataaaataagttatgaaaaaaatcatcatattactaacg 
               
               
                   
                 acatttttcctgctttcgggatgcactattcccagggcggtatttaaatc 
               
               
                   
                 cagccttattaatcaggacgatcctcgttataatctggtcgaagtcacgc 
               
               
                   
                 cgacattaaaactaagcgctcccgatactgtgccgaaaactattgtcgat 
               
               
                   
                 ccggtttttgccgcaaataactggcactggacatctttggctaaaggcga 
               
               
                   
                 tgtgctgcatatcactattttatcctcgggcggggctggatatttatcca 
               
               
                   
                 ataacgcgagcggcgaccgtgcggattttgaaaatattcttgtgactgac 
               
               
                   
                 agtaataccgttcaggtgccttatgccgggacaatcccggtttctggatt 
               
               
                   
                 ggatgtgacgcaactggctgatgagatcaaaaagcgactttcgcgcgttg 
               
               
                   
                 tcctgaatcctcaggtgattgtgacacttaccgcccgcaccggagccatg 
               
               
                   
                 gtgacggtcgagggcagcgggaaaacgggtcgataccctctcgaacagag 
               
               
                   
                 tatgaatcgtctgagtcatcttttggcaacggcagtggcagtagaaaata 
               
               
                   
                 ccagcacagatatgatggaagttcacgtgacgcgccagcagcattatttc 
               
               
                   
                 actgcgcgactgtcagatatttatcagtatcccggattggatattgcctt 
               
               
                   
                 acagccggatgaccgtatcactctgcgtcaagtgaccgagtatgttaatg 
               
               
                   
                 tgctcggcgcagcaggtgttcaggggaaacatgctctggttcagcgtcat 
               
               
                   
                 tccagcgtcgtggatgctttggcgctcgcgaaaggattaaatgacaatct 
               
               
                   
                 tgccgatccgcaagcgatttttctgtacaagcataacgaagcggaacagg 
               
               
                   
                 cgaaacagcagatgcgtaagctgaatatctatcatgtcgatatgagccaa 
               
               
                   
                 cctaactcggtgtttttagcgcaggccatacgagtggacaacggcgatgt 
               
               
                   
                 tatctatatctcgaacgcctctttgactgatttcgctaaggtgaaagccg 
               
               
                   
                 cattcgatagctttttgac 
               
               
                   
               
               
                 SEQ ID NO: 11 (SARS- 
                 RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVL 
               
               
                 CoV-2 RBD) 
                 YNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKI 
               
               
                   
                 ADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDI 
               
               
                   
                 STEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELL 
               
               
                   
                 HAPATVCGPKKSTNLVKNKCVNF 
               
               
                   
               
               
                 SEQ ID NO: 12 (14 
                 GYIPEAPRDGQAYVRKDGEWVLLSTFL 
               
               
                 Fibritin Trimerization 
                   
               
               
                 Motif) 
                   
               
               
                   
               
               
                 SEQ ID NO: 13 (HLYA 
                 ALAYGSQGNLNPLINEISKIISAAGNFDVKEERAAASLLQLSGNASDFSY 
               
               
                 signal sequence) 
                 GRNSITLTASA 
               
               
                   
               
               
                 SEQ ID NO: 14 (YbaS) 
                 MLDANKLQQAVDQAYTQFHSLNGGQNADYIPFLANVPGQLAAVAIVTCDG 
               
               
                   
                 NVYSAGDSDYRFALESISKVCTLALALEDVGPQAVQDKIGADPTGLPFNS 
               
               
                   
                 VIALELHGGKPLSPLVNAGAIATTSLINAENVEQRWQRILHIQQQLAGEQ 
               
               
                   
                 VALSDEVNQSEQTTNFHNRAIAWLLYSAGYLYCDAMEACDVYTRQCSTLI 
               
               
                   
                 NTVELATLGATLAAGGVNPLTHKRVLQADNVPYILAEMMMEGLYGRSGDW 
               
               
                   
                 AYRVGLPGKSGVGGGILAVVPGVMGIAAFSPPLDEEGNSVRGQKMVASVA 
               
               
                   
                 KQLGYNVFKG 
               
               
                   
               
               
                 SEQ ID NO: 15 (YbaS) 
                 ATGTTAGATGCAAACAAATTACAGCAGGCAGTGGATCAGGCTTACACCCA 
               
               
                   
                 ATTTCACTCACTTAACGGCGGACAAAATGCCGATTACATTCCCTTTCTGG 
               
               
                   
                 CGAATGTACCAGGTCAACTGGCGGCAGTGGCTATCGTGACNTGCGATGGC 
               
               
                   
                 AACGTCTATAGTGCGGGTGACAGTGATTACCGCTTTGCACTGGAATCCAT 
               
               
                   
                 CTCTAAAGTCTGTACGTTAGCCCTTGCGTTAGAAGATGTCGGCCCGCAGG 
               
               
                   
                 CGGTACAGGACAAAATTGGCGCTGACCCGACCGGATTGCCCTTTAACTCA 
               
               
                   
                 GTTATCGCCTTAGAGTTGCATGGCGGCAAACCGCTGTCGCCACTGGTAAA 
               
               
                   
                 TGCTGGCGCTATTGCCACCACCAGCCTGATTAACGCTGAAAATGTTGAAC 
               
               
                   
                 AACGCTGGCAGCGAATTTTACATATCCAACAGCAACTGGCTGGTGAGCAG 
               
               
                   
                 GTAGCGCTCTCTGACGAAGTCAACCAGTCGGAACAAACAACCAACTTCCA 
               
               
                   
                 TAACCGGGCCATTGCCTGGCTGCTGTACTCCGCCGGATATCTCTATTGTG 
               
               
                   
                 ATGCAATGGAAGCCTGTGACGTGTACACCCGTCAGTGCTCTACGCTCATC 
               
               
                   
                 AATACTGTTGAACTGGCAACGCTTGGCGCGACGCTGGCGGCGGGTGGTGT 
               
               
                   
                 GAATCCGTTGACGCATAAACGCGTTCTTCAGGCCGACAACGTGCCGTACA 
               
               
                   
                 TTCTGGCCGAAATGATGATGGAAGGGCTGTATGGTCGCTCCGGTGACTGG 
               
               
                   
                 GCGTATCGCGTTGGTTTACCGGGCAAAAGCGGTGTAGGTGGCGGTATTCT 
               
               
                   
                 GGCGGTCGTCCCTGGCGTGATGGGAATTGCCGCGTTCTCACCACCGCTGG 
               
               
                   
                 ACGAAGAAGGCAACAGTGTTCGCGGTCAAAAAATGGTGGCATCGGTCGCT 
               
               
                   
                 AAGCAACTCGGCTATAACGTGTTTAAGGGCTGA 
               
               
                   
               
               
                 SEQ ID NO: 16 (GadB) 
                 MRSNHFKEFKMDKKQVTDLRSELLDSRFGAKSISTIAESKRFPLHEMRDD 
               
               
                   
                 VAFQIINDELYLDGNARQNLATFCQTWDDENVHKLMDLSINKNWIDKEEY 
               
               
                   
                 PQSAAIDLRCVNMVADLWHAPAPKNGQAVGTNTIGSSEACMLGGMAMKWR 
               
               
                   
                 WRKRMEAAGKPTDKPNLVCGPVQICWHKFARYWDVELREIPMRPGQLFMD 
               
               
                   
                 PKRMIEACDENTIGVVPTFGVTYTGNYEFPQPLHDALDKFQADTGIDIDM 
               
               
                   
                 HIDAASGGFLAPFVAPDIVWDFRLPRVKSISASGHKFGLAPLGCGWVIWR 
               
               
                   
                 DEEALPQELVFNVDYLGGQIGTFAINFSRPAGQVIAQYYEFLRLGREGYT 
               
               
                   
                 KVQNASYQVAAYLADEIAKLGPYEFICTGRPDEGIPAVCFKLKDGEDPGY 
               
               
                   
                 TLYDLSERLRLRGWQVPAFTLGGEATDIVVMRIMCRRGFEMDFAELLLED 
               
               
                   
                 YKASLKYLSDHPKLQGIAQQNSFKHT 
               
               
                   
               
               
                 SEQ ID NO: 17 (GadB) 
                 ATGCGATCCAATCATTTTAAGGAGTTTAAAATGGATAAGAAGCAAGTAAC 
               
               
                   
                 GGATTTAAGGTCGGAACTACTCGATTCACGTTTTGGTGCGAAGTCTATTT 
               
               
                   
                 CCACTATCGCAGAATCAAAACGTTTTCCGCTGCACGAAATGCGCGACGAT 
               
               
                   
                 GTCGCATTTCAGATTATCAATGATGAATTATATCTTGATGGCAACGCTCG 
               
               
                   
                 TCAGAACCTGGCCACTTTCTGCCAGACCTGGGACGACGAAAACGTCCACA 
               
               
                   
                 AGTTGATGGATTTATCCATTAACAAAAACTGGATCGACAAAGAAGAATAT 
               
               
                   
                 CCGCAATCCGCAGCCATCGACCTGCGTTGCGTAAACATGGTTGCCGATCT 
               
               
                   
                 GTGGCATGCGCCTGCGCCGAAAAATGGTCAGGCCGTTGGCACCAACACCA 
               
               
                   
                 TTGGTTCTTCCGAGGCCTGTATGCTCGGCGGGATGGCGATGAAATGGCGT 
               
               
                   
                 TGGCGCAAGCGTATGGAAGCTGCAGGCAAACCAACGGATAAACCAAACCT 
               
               
                   
                 GGTGTGCGGTCCGGTGCAAATCTGCTGGCATAAATTCGCCCGCTACTGGG 
               
               
                   
                 ATGTGGAGCTGCGTGAGATCCCTATGCGCCCCGGTCAGTTGTTTATGGAC 
               
               
                   
                 CCGAAACGCATGATTGAAGCCTGCGACGAAAATACCATCGGCGTGGTGCC 
               
               
                   
                 GACTTTCGGCGTGACCTACACCGGTAACTATGAGTTCCCGCAACCGCTGC 
               
               
                   
                 ACGATGCACTGGATAAATTCCAGGCCGACACCGGTATCGACATCGACATG 
               
               
                   
                 CACATCGACGCCGCCAGCGGTGGCTTTCTGGCACCGTTCGTCGCCCCGGA 
               
               
                   
                 TATCGTCTGGGACTTCCGCCTGCCGCGTGTGAAATCGATCAGTGCTTCAG 
               
               
                   
                 GCCATAAATTCGGTCTGGCTCCGCTGGGCTGCGGCTGGGTTATCTGGCGT 
               
               
                   
                 GACGAAGAAGCGCTGCCGCAGGAACTGGTGTTCAACGTTGACTACCTCGG 
               
               
                   
                 TGGTCAGATTGGTACTTTTGCCATCAACTTCTCCCGCCCGGCGGGTCAGG 
               
               
                   
                 TGATTGCACAGTACTATGAATTCCTGCGCCTTGGTCGTGAAGGCTATACC 
               
               
                   
                 AAAGTACAGAATGCTTCCTACCAGGTCGCTGCCTATCTGGCGGATGAGAT 
               
               
                   
                 CGCCAAACTGGGGCCGTATGAGTTCATCTGTACCGGTCGCCCGGACGAAG 
               
               
                   
                 GCATCCCGGCGGTTTGCTTCAAACTGAAAGATGGTGAAGATCCGGGATAC 
               
               
                   
                 ACCCTGTACGACCTCTCTGAACGTCTGCGTCTGCGCGGCTGGCAGGTTCC 
               
               
                   
                 GGCCTTCACTCTCGGCGGTGAAGCCACCGACATCGTGGTGATGCGCATTA 
               
               
                   
                 TGTGTCGTCGCGGCTTCGAAATGGACTTTGCTGAACTGTTGCTGGAAGAC 
               
               
                   
                 TACAAAGCCTCCCTGAAATATCTCAGCGATCACCCGAAACTGCAGGGTAT 
               
               
                   
                 TGCCCAGCAGAACAGCTTTAAACATACCTGA 
               
               
                   
               
               
                 SEQ ID NO: 18 (GadC) 
                 MATSVQTGKAKQLTLLGFFAITASMVMAVYEYPTFATSGFSLVFFLLLGG 
               
               
                   
                 ILWFIPVGLCAAEMATVDGWEEGGVFAWVSNTLGPRWGFAAISFGYLQIA 
               
               
                   
                 IGFIPMLYFVLGALSYILKWPALNEDPITKTIAALIILWALALTQFGGTK 
               
               
                   
                 YTARIAKVGFFAGILLPAFILIALAAIYLHSGAPVAIEMDSKTFFPDFSK 
               
               
                   
                 VGTLVVFVAFILSYMGVEASATHVNEMSNPGRDYPLAMLLLMVAAICLSS 
               
               
                   
                 VGGLSIAMVIPGNEINLSAGVMQTFTVLMSHVAPEIEWTVRVISALLLLG 
               
               
                   
                 VLAEIASWIVGPSRGMYVTAQKNLLPAAFAKMNKNGVPVTLVISQLVITS 
               
               
                   
                 IALIILTNTGGGNNMSFLIALALTVVIYLCAYFMLFIGYIVLVLKHPDLK 
               
               
                   
                 RTFNIPGGKGVKLVVAIVGLLTSIMAFIVSFLPPDNIQGDSTDMYVELLV 
               
               
                   
                 VSFLVVLALPFILYAVHDRKGKANTGVTLEPINSQNAPKGHFFLHPRARS 
               
               
                   
                 PHYIVMNDKKH 
               
               
                   
               
               
                 SEQ ID NO: 19 (GadC) 
                 ATGGCTACATCAGTACAGACAGGTAAAGCTAAGCAGCTCACATTACTCGG 
               
               
                   
                 ATTCTTTGCCATAACGGCATCGATGGTAATGGCTGTTTATGAATACCCTA 
               
               
                   
                 CCTTCGCAACATCGGGCTTTTCATTAGTCTTCTTCCTGCTATTAGGCGGG 
               
               
                   
                 ATTTTATGGTTTATTCCCGTGGGACTTTGTGCTGCGGAAATGGCCACCGT 
               
               
                   
                 CGACGGCTGGGAAGAAGGTGGTGTCTTCGCCTGGGTATCAAATACTCTGG 
               
               
                   
                 GTCCGAGATGGGGATTTGCAGCGATCTCATTTGGCTATCTGCAAATCGCC 
               
               
                   
                 ATTGGTTTTATTCCGATGCTCTATTTCGTGTTAGGGGCACTCTCCTACAT 
               
               
                   
                 CCTGAAATGGCCAGCGCTGAATGAAGACCCCATTACCAAAACTATTGCAG 
               
               
                   
                 CACTCATCATTCTTTGGGCGCTGGCATTAACGCAGTTTGGTGGCACGAAA 
               
               
                   
                 TACACGGCGCGAATTGCTAAAGTTGGCTTCTTCGCCGGTATCCTGTTACC 
               
               
                   
                 TGCATTTATTTTGATCGCATTAGCGGCTATTTACCTGCACTCCGGTGCCC 
               
               
                   
                 CCGTTGCTATCGAAATGGATTCGAAGACCTTCTTCCCTGACTTCTCTAAA 
               
               
                   
                 GTGGGCACACTGGTTGTATTTGTTGCCTTCATTTTGAGTTATATGGGCGT 
               
               
                   
                 AGAAGCTTCCGCAACTCACGTCAATGAAATGAGCAACCCCGGGCGCGACT 
               
               
                   
                 ATCCACTGGCTATGTTACTGCTGATGGTGGCGGCAATCTGCTTAAGCTCT 
               
               
                   
                 GTTGGCGGTTTGTCTATTGCGATGGTCATTCCGGGTAATGAAATCAACCT 
               
               
                   
                 CTCCGCAGGGGTAATGCAAACCTTTACCGTTCTGATGTCCCATGTGGCAC 
               
               
                   
                 CGGAAATTGAGTGGACGGTTCGCGTGATCTCCGCACTGCTGTTGCTGGGT 
               
               
                   
                 GTTCTGGCGGAAATCGCCTCCTGGATTGTTGGTCCTTCTCGCGGGATGTA 
               
               
                   
                 TGTCACAGCGCAGAAAAACCTGCTGCCAGCGGCATTCGCTAAAATGAACA 
               
               
                   
                 AAAATGGCGTACCGGTAACGCTGGTCATTTCGCAGCTGGTGATTACTTCT 
               
               
                   
                 ATCGCGTTGATCATCCTCACCAATACCGGTGGCGGTAACAACATGTCCTT 
               
               
                   
                 CCTGATCGCACTGGCGCTGACGGTGGTGATTTATCTGTGTGCTTATTTCA 
               
               
                   
                 TGCTGTTTATTGGCTACATTGTGTTGGTTCTTAAACATCCTGACTTAAAA 
               
               
                   
                 CGCACATTTAATATCCCTGGTGGTAAAGGGGTGAAACTGGTCGTGGCAAT 
               
               
                   
                 TGTCGGTCTGCTGACTTCAATTATGGCGTTTATTGTTTCCTTCCTGCCGC 
               
               
                   
                 CGGATAACATCCAGGGTGATTCTACCGATATGTATGTTGAATTACTGGTT 
               
               
                   
                 GTTAGTTTCCTGGTGGTACTTGCCCTGCCCTTTATTCTCTATGCTGTTCA 
               
               
                   
                 TGATCGTAAAGGCAAAGCGAATACCGGCGTCACTCTGGAGCCAATCAACA 
               
               
                   
                 GTCAGAACGCACCAAAAGGTCACTTCTTCCTGCACCCGCGTGCACGTTCA 
               
               
                   
                 CCACACTATATTGTGATGAATGACAAGAAACACTAA 
               
               
                   
               
               
                 SEQ ID NO: 20 (araC- 
                 CGATCGTTATGACAACTTGACGGCTACATCATTCACTTTTTCTTCACAAC 
               
               
                 YbaS-GadBC AR 
                 CGGCACGGAACTCGCTCGGGCTGGCCCCGGTGCATTTTTTAAATACCCGC 
               
               
                 cassette) 
                 GAGAAGTAGAGTTGATCGTCAAAACCAACATTGCGACCGACGGTGGCGAT 
               
               
                   
                 AGGCATCCGGGTGGTGCTCAAAAGCAGCTTCGCCTGGCTGATACGTTGGT 
               
               
                   
                 CCTCGCGCCAGCTTAAGACGCTAATCCCTAACTGCTGGCGGAAAAGATGT 
               
               
                   
                 GACAGACGCGACGGCGACAAGCAAACATGCTGTGCGACGCTGGCGATATC 
               
               
                   
                 AAAATTGCTGTCTGCCAGGTGATCGCTGATGTACTGACAAGCCTCGCGTA 
               
               
                   
                 CCCGATTATCCATCGGTGGATGGAGCGACTCGTTAATCGCTTCCATGCGC 
               
               
                   
                 CGCAGTAACAATTGCTCAAGCAGATTTATCGCCAGCAGCTCCGAATAGCG 
               
               
                   
                 CCCTTCCCCTTGCCCGGCGTTAATGATTTGCCCAAACAGGTCGCTGAAAT 
               
               
                   
                 GCGGCTGGTGCGCTTCATCCGGGCGAAAGAACCCCGTATTGGCAAATATT 
               
               
                   
                 GACGGCCAGTTAAGCCATTCATGCCAGTAGGCGCGCGGACGAAAGTAAAC 
               
               
                   
                 CCACTGGTGATACCATTCGCGAGCCTCCGGATGACGACCGTAGTGATGAA 
               
               
                   
                 TCTCTCCTGGCGGGAACAGCAAAATAACACCCGGTCGGCAAACAAATTCT 
               
               
                   
                 CGTCCCTGATTTTTCACCACCCCCTGACCGCGAATGGTGAGATTGAGAAT 
               
               
                   
                 ATAACCTTTCATTCCCAGCGGTCGGTCGATAAAAAAATCGAGATAACCGT 
               
               
                   
                 TGGCCTCAATCGGCGTTAAACCCGCCACCAGATGGGCATTAAACGAGTAT 
               
               
                   
                 CCCGGCAGCAGGGGATCATTTTGCGCTTCAGCCATACTTTTCATACTCCC 
               
               
                   
                 GCCATTCAGAGAAGAAACCAATTGTCCATATTGCATCAGACATTGCCGTC 
               
               
                   
                 ACTGCGTCTTTTACTGGCTCTTCTCGCTAACCAAACCGGTAACCCCGCTT 
               
               
                   
                 ATTAAAAGCATTCTGTAACAAAGCGGGACCAAAGCCATGACAAAAACGCG 
               
               
                   
                 TAACAAAAGTGTCTATAATCACGGCAGAAAAGTCCACATTGATTATTTGC 
               
               
                   
                 ACGGCGTCACACTTTGCTATGCCATAGCATTTTTATCCATAAGATTAGCG 
               
               
                   
                 GATCCTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACCC 
               
               
                   
                 GTTTTTTTGGGAATTCGAGCTCGGAGTTAACAAAAGATGTTAGATGCAAA 
               
               
                   
                 CAAATTACAGCAGGCAGTGGATCAGGCTTACACCCAATTTCACTCACTTA 
               
               
                   
                 ACGGCGGACAAAATGCCGATTACATTCCCTTTCTGGCGAATGTACCAGGT 
               
               
                   
                 CAACTGGCGGCAGTGGCTATCGTGACNTGCGATGGCAACGTCTATAGTGC 
               
               
                   
                 GGGTGACAGTGATTACCGCTTTGCACTGGAATCCATCTCTAAAGTCTGTA 
               
               
                   
                 CGTTAGCCCTTGCGTTAGAAGATGTCGGCCCGCAGGCGGTACAGGACAAA 
               
               
                   
                 ATTGGCGCTGACCCGACCGGATTGCCCTTTAACTCAGTTATCGCCTTAGA 
               
               
                   
                 GTTGCATGGCGGCAAACCGCTGTCGCCACTGGTAAATGCTGGCGCTATTG 
               
               
                   
                 CCACCACCAGCCTGATTAACGCTGAAAATGTTGAACAACGCTGGCAGCGA 
               
               
                   
                 ATTTTACATATCCAACAGCAACTGGCTGGTGAGCAGGTAGCGCTCTCTGA 
               
               
                   
                 CGAAGTCAACCAGTCGGAACAAACAACCAACTTCCATAACCGGGCCATTG 
               
               
                   
                 CCTGGCTGCTGTACTCCGCCGGATATCTCTATTGTGATGCAATGGAAGCC 
               
               
                   
                 TGTGACGTGTACACCCGTCAGTGCTCTACGCTCATCAATACTGTTGAACT 
               
               
                   
                 GGCAACGCTTGGCGCGACGCTGGCGGCGGGTGGTGTGAATCCGTTGACGC 
               
               
                   
                 ATAAACGCGTTCTTCAGGCCGACAACGTGCCGTACATTCTGGCCGAAATG 
               
               
                   
                 ATGATGGAAGGGCTGTATGGTCGCTCCGGTGACTGGGCGTATCGCGTTGG 
               
               
                   
                 TTTACCGGGCAAAAGCGGTGTAGGTGGCGGTATTCTGGCGGTCGTCCCTG 
               
               
                   
                 GCGTGATGGGAATTGCCGCGTTCTCACCACCGCTGGACGAAGAAGGCAAC 
               
               
                   
                 AGTGTTCGCGGTCAAAAAATGGTGGCATCGGTCGCTAAGCAACTCGGCTA 
               
               
                   
                 TAACGTGTTTAAGGGCTGACCATGGAGTGTTTTAATGCGATCCAATCATT 
               
               
                   
                 TTAAGGAGTTTAAAATGGATAAGAAGCAAGTAACGGATTTAAGGTCGGAA 
               
               
                   
                 CTACTCGATTCACGTTTTGGTGCGAAGTCTATTTCCACTATCGCAGAATC 
               
               
                   
                 AAAACGTTTTCCGCTGCACGAAATGCGCGACGATGTCGCATTTCAGATTA 
               
               
                   
                 TCAATGATGAATTATATCTTGATGGCAACGCTCGTCAGAACCTGGCCACT 
               
               
                   
                 TTCTGCCAGACCTGGGACGACGAAAACGTCCACAAGTTGATGGATTTATC 
               
               
                   
                 CATTAACAAAAACTGGATCGACAAAGAAGAATATCCGCAATCCGCAGCCA 
               
               
                   
                 TCGACCTGCGTTGCGTAAACATGGTTGCCGATCTGTGGCATGCGCCTGCG 
               
               
                   
                 CCGAAAAATGGTCAGGCCGTTGGCACCAACACCATTGGTTCTTCCGAGGC 
               
               
                   
                 CTGTATGCTCGGCGGGATGGCGATGAAATGGCGTTGGCGCAAGCGTATGG 
               
               
                   
                 AAGCTGCAGGCAAACCAACGGATAAACCAAACCTGGTGTGCGGTCCGGTG 
               
               
                   
                 CAAATCTGCTGGCATAAATTCGCCCGCTACTGGGATGTGGAGCTGCGTGA 
               
               
                   
                 GATCCCTATGCGCCCCGGTCAGTTGTTTATGGACCCGAAACGCATGATTG 
               
               
                   
                 AAGCCTGCGACGAAAATACCATCGGCGTGGTGCCGACTTTCGGCGTGACC 
               
               
                   
                 TACACCGGTAACTATGAGTTCCCGCAACCGCTGCACGATGCACTGGATAA 
               
               
                   
                 ATTCCAGGCCGACACCGGTATCGACATCGACATGCACATCGACGCCGCCA 
               
               
                   
                 GCGGTGGCTTTCTGGCACCGTTCGTCGCCCCGGATATCGTCTGGGACTTC 
               
               
                   
                 CGCCTGCCGCGTGTGAAATCGATCAGTGCTTCAGGCCATAAATTCGGTCT 
               
               
                   
                 GGCTCCGCTGGGCTGCGGCTGGGTTATCTGGCGTGACGAAGAAGCGCTGC 
               
               
                   
                 CGCAGGAACTGGTGTTCAACGTTGACTACCTCGGTGGTCAGATTGGTACT 
               
               
                   
                 TTTGCCATCAACTTCTCCCGCCCGGCGGGTCAGGTGATTGCACAGTACTA 
               
               
                   
                 TGAATTCCTGCGCCTTGGTCGTGAAGGCTATACCAAAGTACAGAATGCTT 
               
               
                   
                 CCTACCAGGTCGCTGCCTATCTGGCGGATGAGATCGCCAAACTGGGGCCG 
               
               
                   
                 TATGAGTTCATCTGTACCGGTCGCCCGGACGAAGGCATCCCGGCGGTTTG 
               
               
                   
                 CTTCAAACTGAAAGATGGTGAAGATCCGGGATACACCCTGTACGACCTCT 
               
               
                   
                 CTGAACGTCTGCGTCTGCGCGGCTGGCAGGTTCCGGCCTTCACTCTCGGC 
               
               
                   
                 GGTGAAGCCACCGACATCGTGGTGATGCGCATTATGTGTCGTCGCGGCTT 
               
               
                   
                 CGAAATGGACTTTGCTGAACTGTTGCTGGAAGACTACAAAGCCTCCCTGA 
               
               
                   
                 AATATCTCAGCGATCACCCGAAACTGCAGGGTATTGCCCAGCAGAACAGC 
               
               
                   
                 TTTAAACATACCTGATAACCGTTTTGGAGTGATAATATGGCTACATCAGT 
               
               
                   
                 ACAGACAGGTAAAGCTAAGCAGCTCACATTACTCGGATTCTTTGCCATAA 
               
               
                   
                 CGGCATCGATGGTAATGGCTGTTTATGAATACCCTACCTTCGCAACATCG 
               
               
                   
                 GGCTTTTCATTAGTCTTCTTCCTGCTATTAGGCGGGATTTTATGGTTTAT 
               
               
                   
                 TCCCGTGGGACTTTGTGCTGCGGAAATGGCCACCGTCGACGGCTGGGAAG 
               
               
                   
                 AAGGTGGTGTCTTCGCCTGGGTATCAAATACTCTGGGTCCGAGATGGGGA 
               
               
                   
                 TTTGCAGCGATCTCATTTGGCTATCTGCAAATCGCCATTGGTTTTATTCC 
               
               
                   
                 GATGCTCTATTTCGTGTTAGGGGCACTCTCCTACATCCTGAAATGGCCAG 
               
               
                   
                 CGCTGAATGAAGACCCCATTACCAAAACTATTGCAGCACTCATCATTCTT 
               
               
                   
                 TGGGCGCTGGCATTAACGCAGTTTGGTGGCACGAAATACACGGCGCGAAT 
               
               
                   
                 TGCTAAAGTTGGCTTCTTCGCCGGTATCCTGTTACCTGCATTTATTTTGA 
               
               
                   
                 TCGCATTAGCGGCTATTTACCTGCACTCCGGTGCCCCCGTTGCTATCGAA 
               
               
                   
                 ATGGATTCGAAGACCTTCTTCCCTGACTTCTCTAAAGTGGGCACACTGGT 
               
               
                   
                 TGTATTTGTTGCCTTCATTTTGAGTTATATGGGCGTAGAAGCTTCCGCAA 
               
               
                   
                 CTCACGTCAATGAAATGAGCAACCCCGGGCGCGACTATCCACTGGCTATG 
               
               
                   
                 TTACTGCTGATGGTGGCGGCAATCTGCTTAAGCTCTGTTGGCGGTTTGTC 
               
               
                   
                 TATTGCGATGGTCATTCCGGGTAATGAAATCAACCTCTCCGCAGGGGTAA 
               
               
                   
                 TGCAAACCTTTACCGTTCTGATGTCCCATGTGGCACCGGAAATTGAGTGG 
               
               
                   
                 ACGGTTCGCGTGATCTCCGCACTGCTGTTGCTGGGTGTTCTGGCGGAAAT 
               
               
                   
                 CGCCTCCTGGATTGTTGGTCCTTCTCGCGGGATGTATGTCACAGCGCAGA 
               
               
                   
                 AAAACCTGCTGCCAGCGGCATTCGCTAAAATGAACAAAAATGGCGTACCG 
               
               
                   
                 GTAACGCTGGTCATTTCGCAGCTGGTGATTACTTCTATCGCGTTGATCAT 
               
               
                   
                 CCTCACCAATACCGGTGGCGGTAACAACATGTCCTTCCTGATCGCACTGG 
               
               
                   
                 CGCTGACGGTGGTGATTTATCTGTGTGCTTATTTCATGCTGTTTATTGGC 
               
               
                   
                 TACATTGTGTTGGTTCTTAAACATCCTGACTTAAAACGCACATTTAATAT 
               
               
                   
                 CCCTGGTGGTAAAGGGGTGAAACTGGTCGTGGCAATTGTCGGTCTGCTGA 
               
               
                   
                 CTTCAATTATGGCGTTTATTGTTTCCTTCCTGCCGCCGGATAACATCCAG 
               
               
                   
                 GGTGATTCTACCGATATGTATGTTGAATTACTGGTTGTTAGTTTCCTGGT 
               
               
                   
                 GGTACTTGCCCTGCCCTTTATTCTCTATGCTGTTCATGATCGTAAAGGCA 
               
               
                   
                 AAGCGAATACCGGCGTCACTCTGGAGCCAATCAACAGTCAGAACGCACCA 
               
               
                   
                 AAAGGTCACTTCTTCCTGCACCCGCGTGCACGTTCACCACACTATATTGT 
               
               
                   
                 GATGAATGACAAGAAACACTAACGATCG 
               
               
                   
               
               
                 SEQ ID NO: 21 (GadA) 
                 MDQKLLTDFRSELLDSRFGAKAISTIAESKRFPLHEMRDDVAFQIINDEL 
               
               
                   
                 YLDGNARQNLATFCQTWDDENVHKLMDLSINKNWIDKEEYPQSAAIDLRC 
               
               
                   
                 VNMVADLWHAPAPKNGQAVGTNTIGSSEACMLGGMAMKWRWRKRMEAAGK 
               
               
                   
                 PTDKPNLVCGPVQICWHKFARYWDVELREIPMRPGQLFMDPKRMIEACDE 
               
               
                   
                 NTIGVVPTFGVTYTGNYEFPQPLHDALDKFQADTGIDIDMHIDAASGGFL 
               
               
                   
                 APFVAPDIVWDFRLPRVKSISASGHKFGLAPLGCGWVIWRDEEALPQELV 
               
               
                   
                 FNVDYLGGQIGTFAINFSRPAGQVIAQYYEFLRLGREGYTKVQNASYQVA 
               
               
                   
                 AYLADEIAKQGPYEFICTGRPDEGIPAVCFKLKDGEDPGYTLYDLSERLR 
               
               
                   
                 LRGWQVPAFTLGGEATDIVVMRIMCRRGFEMDFAELLLEDYKASLKYLSD 
               
               
                   
                 HPKLQGIAQQNSFKHT 
               
               
                   
               
               
                 SEQ ID NO: 22 (GadA) 
                 ATGGACCAGAAGCTGTTAACGGATTTCCGCTCAGAACTACTCGATTCACG 
               
               
                   
                 TTTTGGCGCAAAGGCCATTTCTACTATCGCGGAGTCAAAACGATTTCCGC 
               
               
                   
                 TGCACGAAATGCGCGATGATGTCGCATTCCAGATTATCAATGATGAATTA 
               
               
                   
                 TATCTTGATGGCAACGCTCGTCAGAACCTAGCCACTTTCTGCCAGACCTG 
               
               
                   
                 GGACGACGAAAACGTCCATAAATTGATGGATTTGTCGATCAATAAAAACT 
               
               
                   
                 GGATCGACAAAGAAGAATATCCGCAATCCGCAGCCATCGACCTGCGTTGC 
               
               
                   
                 GTAAATATGGTTGCCGATCTGTGGCATGCGCCTGCGCCGAAAAATGGTCA 
               
               
                   
                 GGCCGTTGGCACCAACACCATTGGTTCTTCCGAGGCCTGTATGCTCGGCG 
               
               
                   
                 GGATGGCGATGAAATGGCGTTGGCGCAAGCGTATGGAAGCTGCAGGCAAA 
               
               
                   
                 CCAACGGATAAACCAAACCTAGTGTGCGGTCCGGTACAAATCTGCTGGCA 
               
               
                   
                 TAAATTCGCCCGCTACTGGGATGTGGAGCTGCGTGAGATCCCTATGCGCC 
               
               
                   
                 CCGGTCAGTTGTTTATGGACCCGAAACGCATGATTGAAGCCTGTGACGAA 
               
               
                   
                 AACACCATCGGCGTGGTGCCGACTTTCGGCGTGACCTACACCGGTAACTA 
               
               
                   
                 TGAGTTCCCACAACCGCTGCACGATGCGCTGGATAAATTCCAGGCCGACA 
               
               
                   
                 CCGGTATCGACATCGACATGCACATCGACGCTGCCAGCGGTGGCTTCCTG 
               
               
                   
                 GCACCGTTCGTCGCCCCGGATATCGTCTGGGACTTCCGCCTGCCGCGTGT 
               
               
                   
                 GAAATCGATCAGTGCTTCAGGCCATAAATTCGGTCTGGCTCCGCTGGGCT 
               
               
                   
                 GCGGCTGGGTTATCTGGCGTGACGAAGAAGCGCTGCCGCAGGAACTGGTG 
               
               
                   
                 TTCAACGTTGACTACCTGGGTGGTCAAATTGGTACTTTTGCCATCAACTT 
               
               
                   
                 CTCCCGCCCGGCGGGTCAGGTAATTGCACAGTACTATGAATTCCTGCGCC 
               
               
                   
                 TCGGTCGTGAAGGCTATACCAAAGTACAGAACGCCTCTTACCAGGTTGCC 
               
               
                   
                 GCTTATCTGGCGGATGAAATCGCCAAACAGGGGCCGTATGAGTTCATCTG 
               
               
                   
                 TACGGGTCGCCCGGACGAAGGCATCCCGGCGGTTTGCTTCAAACTGAAAG 
               
               
                   
                 ATGGTGAAGATCCGGGATACACCCTGTACGACCTCTCTGAACGTCTGCGT 
               
               
                   
                 CTGCGCGGCTGGCAGGTTCCGGCCTTCACTCTCGGCGGTGAAGCCACCGA 
               
               
                   
                 CATCGTGGTGATGCGCATTATGTGTCGTCGCGGCTTCGAAATGGACTTTG 
               
               
                   
                 CTGAACTGTTGCTGGAAGACTACAAAGCCTCCCTGAAATATCTCAGCGAT 
               
               
                   
                 CACCCGAAACTGCAGGGTATTGCCCAGCAGAACAGCTTTAAACACACCTG 
               
               
                   
                 A