LOCUS       40924_46233        7375 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pWB206, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7375)
  AUTHORS   Bourn WR, Shephard E, Powels R, Stutz H, Douglas N, Jaffray A, 
            Williamson A-L.
  TITLE     The use of guided evolution to create a stable, immunogenic HIV-1 
            antigen expressing recombinant BCG
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7375)
  AUTHORS   Shephard E, Bourn WR, Stutz H, Johnston N, van Harmelen JH, 
            Williamson C, Douglass N, Bowers D, Galant S, Isaacs Z, Binder A, 
            Williamson A-L.
  TITLE     Priming with a rBCG or DNA vaccine expressing HIV-I subtype C Gag 
            elicits comparable CD8+ T cell responses when boosted by a rMVA 
            expressing GagC
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 7375)
  AUTHORS   Bourn WR.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JAN-2007) Medical Virology and Institute of Infectious
            Disease and Molecular Medicine, University of Cape Town, Anzio Road 
            Observatory, Cape Town 7925, South Africa
REFERENCE   4  (bases 1 to 7375)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7375)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (10-JAN-2007) Medical Virology and Institute of Infectious Disease 
            and Molecular Medicine, University of Cape Town, Anzio Road 
            Observatory, Cape Town 7925, South Africa"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             120..932
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1267..1855
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      2215..4799
                     /note="mycobacterial origin; OriM"
     primer_bind     4829..4845
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      4857..4943
                     /label=BCG hsp60
                     /note="BCG hsp60"
                     /regulatory_class="promoter"
     regulatory      4967..4973
                     /note="strong synthetic Shine Delgarno sequence; RBSA"
                     /regulatory_class="ribosome_binding_site"
     gene            5010..5372
                     /gene="tat"
                     /label=tat
                     /note="shuffled BCG codon optimized tat gene from HIV-1
                     subtype C isolate"
     gene            5382..5804
                     /gene="gag"
                     /label=gag
                     /note="truncated fragment of BCG codon optimized gag gene
                     from HIV-1 subtype C isolate"
     regulatory      5689..5695
                     /gene="gag"
                     /label=fortuitous RBSD
                     /note="fortuitous RBSD"
                     /regulatory_class="ribosome_binding_site"
     gene            5807..6460
                     /gene="gag"
                     /label=gag
                     /note="truncated fragment of BCG codon optimized gag gene
                     from HIV-1 subtype C isolate"
     misc_feature    6518..6547
                     /label=Gag-GFP linker sequence
                     /note="Gag-GFP linker sequence"
     CDS             6545..7258
                     /label=Cycle 3 GFP
                     /note="Cycle 3 GFP (Crameri et al., 1996)"
     terminator      7293..7339
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 gctagcaaca aagcgacgtt gtgtctcaaa atctctgatg ttacattgca caagataaaa
       61 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
      121 tgagccatat tcaacgggaa acgtcttgct cgaggccgcg attaaattcc aacatggatg
      181 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
      241 atcgcttgta tgggaagccc catgcgccag agttgtttct gaaacatggc aaaggtagcg
      301 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
      361 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
      421 tccccgggaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
      481 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
      541 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
      601 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
      661 aaatgcataa tcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
      721 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
      781 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
      841 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
      901 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
      961 actggcagag cattacgctg acttgacggg acggcggctt tgttgaataa atcgaacttt
     1021 tgctgagttg aaggatcaga tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa
     1081 gcaaaagttc aaaatcacca actggtccac ctacaacaaa gctctcatca accgtggctc
     1141 cctcactttc tggctggatg atggggcgat tcaggcctgg tatgagtcag caacaccttc
     1201 ttcacgaggc agacctcact agttccactg agcgtcagac cccgtagaaa agatcaaagg
     1261 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1321 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     1381 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     1441 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     1501 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     1561 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     1621 aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc
     1681 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     1741 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1801 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1861 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     1921 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     1981 cgctcgccgc agccgaacga ccgagcgcaa cgcgggccgc ccggagcccc gcccgcgctc
     2041 gggagccccg ggcccgcgcc gaagcccgct gctgcgagcc cggagcgggc cggccgacgg
     2101 cggtgcgggc ccggcggcgg acgctcagca gcggcgggcg tgaaaggccc tggcatcctc
     2161 gatcatctcc tccagggtgg tcggccgagc ttccatccca gctcggcaag gatcaacgag
     2221 gacagtcgca cgacgaagtt cttctggatc gcgcccgtgc tggaagcact caacctcgaa
     2281 gcgtgtggtt gcggagccat ctagcaacca cacgaaacat gcgcaacgaa ccgcgcaacg
     2341 aacaacgcct agaactggcc ctagatgagc tgactcgtat cgttggtaaa cctagtttga
     2401 ccagcatgtt ttaactacgt tcggtgagct gtcaacgggg cctgtaacgg cacaacgaac
     2461 cgtgcaacga gagtggccac ggatgccacc acaggcacta caacggagtt cgccacgtac
     2521 atcaccacaa ccaccgattc tggcggtgag ctccccgata ttcagcggaa atggcttggt
     2581 atcgaccaag attcgtagaa ccccgtctcg tctggctggt attcaaaacg ggcgcaacga
     2641 aacacgcaac gagacaggca tggcccaaac cagaaaacta gcgtctacca ggacttttac
     2701 ctgtccgacc cgttgcaacg gaacccccca cggaaccccc gcgacacccg ctccccaatt
     2761 gcgttagaac agcggtggat tgtcggcttc gttgtgggcc ttttgagccg cttcctgttc
     2821 tgccgcacgc tctttcctcg cccgatagcc gagtcgctta acggtgtcca gatgcagccc
     2881 gaaatgtttg gccgtttgcg gccaagagtg gccctcgtcg tcgtgatagg cgcggatgcg
     2941 ttcgcggcgt gcagcctgct cggcgagcca ctcgctgcgt tcctgcgcca cgagccggac
     3001 gacgtggcgt tcggatagtc cggtgattcg agcgccttcg gcggcggtca cgcgccgctt
     3061 tttgcggaca gtcggctgcc ggttgtagcc gtcgctgtag ccgtcgctgt agccgtcgct
     3121 catagcaatg cctccatggc tgacgcggac tttgcgcgcc gcgcaactgt gctcgccgcc
     3181 gtgcgcgctg ctcccttccg cgagatggcc gactggcgcg cactgagtgt ggcctcgtag
     3241 accacgatcc cgtccgccca aatgcgcgac ttggttgtga tccaacgcca aatgctgttg
     3301 gcgatggcgc ggacctcgct gtccggtagc ggtccgggac acacgtcgtt gcacgggaat
     3361 tcggcgtttc gcgcgtggca ctcggcatag atcgcgcggc cgagtccgtc cacgttccgg
     3421 gtcggcaggt agatccgcat gagggcggga cgataggccc acaacctgac ggaatcgaac
     3481 agtgcgcaat tccgccctag cggcgtcgga gccgctttgt acgtggtctg ctgacgccag
     3541 cgcggcggtg gcatgttcgc gccgagctcg gcctcgatgt ggctgagtgt gtagagatct
     3601 gagtggagcc attccgtttc ccaggcgatg tggccggggt ttttggtcat gaggcctgag
     3661 taactgcggt cgccgtcgac ggcgcgccga aggccttcgg cgcacgccgc catgtatgcg
     3721 agcggcttac gccgcgcgta ttcggtgcgt ggaacagggg cgttgagtgc ccacactgcg
     3781 tgtgcgtggc cgttggcgcg attgcccacg atcgcgttgg gcagcggatg ggacccccgg
     3841 gcgctgagcg ctcggagcgc tgcgtctgga tggtctacgt ccacgaccag caggtttgcc
     3901 agcgctgttg ggttcgcctc gatgtaccgg cggcctaggg ccgacgcgcg gctttggcgg
     3961 tagatcccct cgagcagatc gtcgcttgcc agcggccagt acggcagcca gagctgctca
     4021 aattcgtcgg cgacgtggct cacgcttggt agtagaccac gattaatcac cggtgtatgg
     4081 tccgacacga gctccaagtc agatatttcg ctgaggggcc accccacaac tgcacactcc
     4141 cccgctctcc cgtcgagccc tggtggtgga acaccagcga cagccgagca cccccaacca
     4201 cctgtaccaa ccaggaggaa cacatgcgtc gtttcgagga cgtttccggg ccgctgagag
     4261 ccgctgtggc ggccgtacac gccgccttag acccgttaga ccccctgccg cctgaatgcg
     4321 cgggtacgag ccacacagcg cccgaactta cggagctggt gggctcacct ggctttatgg
     4381 cgtacgaatc ggctgtgtgc gacctgttgg gcgaggtgag gtacgcgcta ctcacgctgg
     4441 caagggcgac acagccgccc caccgagccc gcacggccgc gcgcggtgtc aacaaccggg
     4501 tgagtcgtgc acaccagcag gtgttcgagg cttggctcga agtgcaggac atcgtggcga
     4561 acgccgcccg atgagccgcg ccttacgctg gctgccagcc gttcgcgggc tggttggtgc
     4621 agcgcgtcga gcggttagag gccctgcggt gttccaccac cgcaggcctc gcccttttta
     4681 aggctgaatt tgcttgtctc cgaatccaac tggcttgtcc aagggtgtat ctacgcttaa
     4741 tccaaagttc aaacgagggg attacacatg accaacttcg ataacgttct cggctcgatg
     4801 atgcccgggt acgtaccggg ccccccctcg aggtcgacgg tatcgataag cttgatctag
     4861 acaaggtcga acgaggggca tgacccggtg cggggcttct tgcactcggc ataggcgagt
     4921 gctaagaata acgttggcac tcgaattcga gttaggtacg tgacttaagg aggcaaaatt
     4981 ccatgggtta cgtatcgtcg acccaagctc tgggcatctc gtacggccgc aagaagcgcc
     5041 gccagcgccg ctcgaccccg ccgtcgtcgg aggaccacca gaacccgatc tcgaagcagc
     5101 cgctgccgca gacccgcggc gacccgaccg gctcggagga gtcgaagaag aaggtcgaga
     5161 gcaagaccaa gaccgacccg ttcgactgca agtactgctc gtaccactgc ctggtgtgct
     5221 tccagaccaa gggcctcggt atctcctacg ggcggaagaa gcggatggag cccatcgacc
     5281 cgaacctgga gccctggaac cacccgggct cgcagccgaa caccccgtgc aacaagtgct
     5341 attgcaaata ctgctcctat cactgcctcg tcgtcgaagc tgcccgcgcc tcgatcctgc
     5401 gcggcgagaa gctggacaag tgggagaaga tccgcctgcg cccgggcggc aagaagcact
     5461 acatgctgaa gcacatcgtg tgggcctcgc gcgagctgga gcgcttcgcc ctgaacccgg
     5521 acctgctgga gacctcggag ggctgcaagc agatcatgaa gcagctccag ccggccctcc
     5581 agaccggcac cgagaagctc aagtcgctgt acaacaccgt ggccaccctg tactgcgtgc
     5641 acgagaagat cgaggtgcgc gacaccaagg aggccctgga caagatcgag gaggagcaga
     5701 acaagtgcca gcaaaagacc cagcaggcca aggccgccga cggcaaggtg tcgcagaact
     5761 acccgattat gcagaacctc cagggccaga tggtgcacca ggcctctcgc cgcgcaccct
     5821 gaacgcctgg gtgaaggtga tcgaggagaa ggcgttctcg ccggaggtga tcccgatgtt
     5881 caccgccctg tcggagggtg ccaccccgca ggacctgaac accatgctga acaccgtggg
     5941 cggccaccag gccgccatgc agatgctgaa ggacaccatc aacgaggagg ccgccgagtg
     6001 ggaccgcctg cacccggtgc acgccggccc gattgccccg ggccagatgc gcgagccgcg
     6061 cggctcggac atcgccggca ccacctcgac cctccaggag cagatcgcct ggatgacctc
     6121 gaacccgccg atcccggtgg gcgacatcta caagcgctgg atcatcctgg gcctgaacaa
     6181 gatcgtgcgc atgtactcgc cggtgtcgat cctggacatc cgccagggtc cgaaggagcc
     6241 gttccgcgac tacgtggacc gcttcttcaa gaccctgcgc gccgagcagg ccacccagga
     6301 ggtgaagaac tggatgaccg acaccctgct ggtgcagaac gccaacccgg actgcaagac
     6361 catcctgcgc gccctgggtc cgggtgccac cctggaggag atgatgaccg cctgccaggg
     6421 cgtgggcggc ccgggccaca aggcccgcgt gctggccgaa ggccatgtcg cagaccaact
     6481 cgggcaacat catgatgcag cgctcgaact tcaagggtcc ggctccgccg gttccgccgg
     6541 ctcagcttcg aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga
     6601 tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata
     6661 cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac
     6721 acttgtcact actttctctt atggtgttca atgcttttca agatacccag atcatatgaa
     6781 gcggcacgac ttcttcaaga gcgccatgcc tgagggatac gtgcaggaga ggaccatctc
     6841 tttcaaggac gacgggaact acaagacacg tgctgaagtc aagtttgagg gagacaccct
     6901 cgtcaacagg atcgagctta agggaatcga tttcaaggag gacggaaaca tcctcggcca
     6961 caagttggaa tacaactaca actcccacaa cgtatacatc acggcagaca aacaaaagaa
     7021 tggaatcaaa gctaacttca aaattagaca caacattgaa gatggaagcg ttcaactagc
     7081 agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca
     7141 ttacctgtcc acacaatctg ccctttcgaa agatcccaac gaaaagagag accacatggt
     7201 ccttcttgag tttgtaacag ctgctgggat tacacatggc atggatgaac tatacaaatg
     7261 agttaactag cgtacgatcg actgccaggc atcaaataaa acgaaaggct cagtcgaaag
     7321 actgggcctt tcgttttatc tgttgtttgt ccggccatca tggccgcggt gatca
//
