LOCUS       V009528                 7366 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009528
VERSION     V009528
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7366)
  AUTHORS   Fortune SM, Jaeger A, Sarracino DA, Chase MR, Sassetti CM, Sherman
            DR, Bloom BR, Rubin EJ.
  TITLE     Mutually dependent secretion of proteins required for mycobacterial
            virulence
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 102 (30), 10676-10681 (2005)
   PUBMED   16030141
REFERENCE   2  (bases 1 to 7366)
  AUTHORS   Fortune SM, Jaeger A, Sherman DR, Rubin EJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-JUN-2005) IID, Harvard School of Public Health, 200
            Longwood Ave., Boston, MA 02115, USA
REFERENCE   3  (bases 1 to 7366)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7366)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
            Acad. Sci. U.S.A."; date: "2005"; volume: "102"; issue: "30"; pages:
            "10676-10681"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (03-JUN-2005) IID, Harvard School of Public Health, 200 Longwood
            Ave., Boston, MA 02115, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      52..138
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      230..257
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        303..333
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(339..358)
                     /label="lac operator (symmetric)"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The
                     symmetric lac operator was optimized for tight binding of
                     lac repressor."
     CDS             387..1562
                     /gene="espA"
                     /label="ESX-1 secretion-associated protein EspA"
                     /note="ESX-1 secretion-associated protein EspA from
                     Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv).
                     Accession#: P9WJE1"
     CDS             1569..1586
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             1593..1622
                     /label="Myc"
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     terminator      1666..1713
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     rep_origin      complement(1917..2505)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(2827..3483)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(3484..3584)
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             complement(3582..4547)
                     /gene="hyg"
                     /label="Hygromycin-B 7''-O-kinase"
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces
                     hygroscopicus. Accession#: P09979"
     CDS             complement(5743..6855)
                     /gene="33"
                     /label="Integrase"
                     /note="Integrase from Mycobacterium phage L5. Accession#:
                     P22884"
ORIGIN
        1 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
       61 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
      121 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
      181 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
      241 ctgacggatg gcctttttgc gtttctacaa actcttcctg tcgtcatatc tagaccccag
      301 gcttgacact ttatgcttcc ggctcgtata atgtgtggaa ttgtgagcgc tcacaattcg
      361 gatccagctg cagaattcgg ccggccatga gcagagcgtt catcatcgat ccaacgatca
      421 gtgccattga cggcttgtac gaccttctgg ggattggaat acccaaccaa gggggtatcc
      481 tttactcctc actagagtac ttcgaaaaag ccctggagga gctggcagca gcgtttccgg
      541 gtgatggctg gttaggttcg gccgcggaca aatacgccgg caaaaaccgc aaccacgtga
      601 attttttcca ggaactggca gacctcgatc gtcagctcat cagcctgatc cacgaccagg
      661 ccaacgcggt ccagacgacc cgcgacatcc tggagggcgc caagaaaggt ctcgagttcg
      721 tgcgcccggt ggctgtggac ctgacctaca tcccggtcgt cgggcacgcc ctatcggccg
      781 ccttccaggc gccgttttgc gcgggcgcga tggccgtagt gggcggcgcg cttgcctact
      841 tggtcgtgaa aacgctgatc aacgcgactc aactcctcaa attgcttgcc aaattggcgg
      901 agttggtcgc ggccgccatt gcggacatca tttcggatgt ggcggacatc atcaagggca
      961 ccctcggaga agtgtgggag ttcatcacaa acgcgctcaa cggcctgaaa gagctttggg
     1021 acaagctcac ggggtgggtg accggactgt tctctcgagg gtggtcgaac ctggagtcct
     1081 tctttgcggg cgtccccggc ttgaccggcg cgaccagcgg cttgtcgcaa gtgactggct
     1141 tgttcggtgc ggccggtctg tccgcatcgt cgggcttggc tcacgcggat agcctggcga
     1201 gctcagccag cttgcccgcc ctggccggca ttgggggcgg gtccggtttt gggggcttgc
     1261 cgagcctggc tcaggtccat gccgcctcaa ctcggcaggc gctacggccc cgagctgatg
     1321 gcccggtcgg cgccgctgcc gagcaggtcg gcgggcagtc gcagctggtc tccgcgcagg
     1381 gttcccaagg tatgggcgga cccgtaggca tgggcggcat gcacccctct tcgggggcgt
     1441 cgaaagggac gacgacgaag aagtactcgg aaggcgcggc ggcgggcact gaagacgccg
     1501 agcgcgcgcc agtcgaagct gacgcgggcg gtgggcaaaa ggtgctggta cgaaacgtcg
     1561 tcactagtca ccaccaccac caccatatgg ccgagcagaa gctgatctcg gaggaggacc
     1621 tgtgacttaa gcttatcgat gtcgacgtag ttaactgagc aataactagc ataacccctt
     1681 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggagat
     1741 ctggtaccgc ggccgcacgc gttgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1801 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1861 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1921 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1981 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2041 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2101 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2161 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2221 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2281 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2341 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2401 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2461 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2521 ttttctacgg ggtctgacgc tcagtggaac tagtgaggtc tgcctcgtga agaaggtgtt
     2581 gctgactcat accaggcctg aatcgcccca tcatccagcc agaaagtgag ggagccacgg
     2641 ttgatgagag ctttgttgta ggtggaccag ttggtgattt tgaacttttg ctttgccacg
     2701 gaacggtctg cgttgtcggg aagatgcgtg atctgatcct tcaactcagc aaaagttcga
     2761 tttattcaac aaagccgccg tcccgtcaag tcagcgtaat gctctgccag tgttacaacc
     2821 aagttacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
     2881 gacatggaag ccatcacaga cggcatgatg aacctgaatc gccagcggca tcagcacctt
     2941 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
     3001 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
     3061 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     3121 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     3181 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     3241 ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat
     3301 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     3361 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     3421 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     3481 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     3541 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgaggaactg cgccagttcc
     3601 tccggatcgg tgaagccgga gagatccagc ggggtctcct cgaacacctc gaagtcgtgc
     3661 aggaaggtga aggcgagcag ttcgcgggcg aagtcctcgg tccgcttcca ctgcgccccg
     3721 tcgagcagcg cggccaggat ctcgcggtcg ccccggaagg cgttgagatg cagttgcacc
     3781 aggctgtagc gggagtctcc cgcatagacg tcggtgaagt cgacgatccc ggtgacctcg
     3841 gtcgcggcca ggtccacgaa gatgttggtc ccgtgcaggt cgccgtggac gaaccggggt
     3901 tcgcggccgg ccagcagcgt gtccacgtcc ggcagccagt cctccaggcg gtccagcagc
     3961 cggggcgaga ggtagcccca cccgcggtgg tcctcgacgg tcgccgcgcg gcgttcccgc
     4021 agcagttccg ggaagacctc ggaatggggg gtgagcacgg tgttcccggt cagcggcacc
     4081 ctgtgcagcc ggccgagcac ccggccgagt tcgcgggcca gggcgagcag cgcgttccgg
     4141 tcggtcgtgc cgtccatcgc ggaccgccag gtggtgccgg tcatccggct catcaccagg
     4201 tagggccacg gccaggctcc ggtgccgggc cgcagctcgc cgcggccgag gaggcggggc
     4261 accggcaccg gggcgtccgc caggaccgcg tacgcctccg actccgacgc gaggctctcc
     4321 ggaccgcacc agtgctcgcc gaacagcttg atcaccgggt cgggctcgcc gaccagtacg
     4381 gggttggtgc tctcgccggg cacccgcagc accggcggca ccggcagccc gagctcctcc
     4441 agggctcggc gggccagcgg ctcccagaat tcctggtcgt tccgcaggct cgcgtaggaa
     4501 tcatccgaat caatacggtc gagaagtaac agggattctt gtgtcacagc ggacctctat
     4561 tcacagggta cgggccggct taattccgca cggccggtcg cgacacggcc tgtccgcacc
     4621 gcggtcaggc gttgacgatg acgggctggt cggccacgtc ggggagtata acttcctcac
     4681 acaaccggtc gtgactttta gggctccgag agaagctcct cgatgtcgtc tggccacgac
     4741 cagaggagtt caccctcggc ggtgaggttg gtgtgctcgt tcacccggat caggagatcg
     4801 tcatcctcga tgcctcgggg gacgtacctg aacccgccgc cggccatacc ttcgtagggc
     4861 tcgatggatg ggtcgaactc gagcactaag tcgtcgtcgc ggagcatctt ccaccacgac
     4921 aataggcgct tcttcttgtc ttcggacatc gtgcggaagc tacccactcg catgtactcg
     4981 ccgtgatccc ggagcctctg aaaagccttc gacttatcgt gaggtttccg cgtgtcccac
     5041 ggccagttct gctggacgat ctgcctggtg gtcaaccgtc ctccgtaggt cttcttgtgc
     5101 cacgacaccg cttgtcgagt cacgccatac agctctgcga tttcggtctg attaaacccc
     5161 ttcctgcgaa gatcttcgat ctcgctgaga gtgagtggta ttcggctagg ggccggaacc
     5221 actgctttgt gttggatttt gccgctcatg tttccctcca tgagaaaggt gcgtgcgtct
     5281 ccgccgatta cggagacatg ttggtgcctg tcaaggatac ccctaattta gttgcgtctg
     5341 cggaaccata ttcagttgtg ttccccgacg ccgtggccgt ctcccactgg gcgtgggatc
     5401 gactggcgtt acgcggtcgt aaatgtagcg gcctgcccca ctcggtagca aaccttgtga
     5461 caggtatcac ttaggtcgcc ttctgttaca cgttgacctc gggtttcatc gtcacgactc
     5521 tcctttctta gacagcctca agatcgttac accggcttgc gaagatgtac cttctagctg
     5581 gcgggaacgg caatgctcct ggcggcggcg gggctggtgc ccaggtctcc gcacagagcg
     5641 gcggtgctgg cgctcgcggc caggcgtggt tcttcgcgta ctgacaagaa acccccctct
     5701 ttaggactca gtgtccttgg gaggggggct ttttgcgttt caggaggtct tggccagctt
     5761 ggacatcgcc tcagcgatag cctcgtcgcg ggcctcagac gccatctggt acttcatcgc
     5821 catcctagga gtcgtgtgac cgagacgggc catcagctcc ttggtcgtcg cacctgcctg
     5881 agcggcgaac gtagcgccga cagcgcggag gtcgtggatg cggagttccg gccgaccgat
     5941 cttggcgtag ccacgcttca gcgacttggt gaacgcggac ttcgacagcc ggttgccctg
     6001 cgtcgtggtc accaggaatg cctcggggcc cttgttcatc ttcgtacggt ccttcatgtg
     6061 cgctcggatc atctccgcga cgtgaggcgg aaccgtcaca ggacgcttcg accggacggt
     6121 cttggcgttg ccaacgacga tcttgttccc cacgcgggaa gcgccacggc gcacccggag
     6181 cttcatcgtc atgccgtcgt gcacgatgtc cttgcggcga agctcgatca gctctccgaa
     6241 ccggaggctc gtccacgcca ggatgtatgc cgcgatccgg tagtgctcga agatctcagc
     6301 ggcgacgatg tccagctcct caggcgtcag cgcctctacg tcgcgctcat cggctgcctt
     6361 ctgctcgatc cggcacgggt tctctgcgat cagcttgtcc tcgaccgctg tgttcatcac
     6421 cgcccggagg acgttgtagg catgccggcg ggcagtcggg tgcttcctac ccatcccggc
     6481 ccaccacgca cgcaccagag ctggcgtcat ctctgtgacc gccacttcac ctagcaccgg
     6541 gtagatgcgg cgctccgcgt gcccgctgta cagatccctg gtgccgtctg cgaggtcgcg
     6601 ctccacgagc cacttccggg tgtactcctc cagcgtgatg gcgctggcgg ctgccttctt
     6661 cgcccggtcc tgtggagggg tccaggtctc catctcgatg agccgcttct cgcccgcgag
     6721 ccaggcttcg gcgtccatct tgttgtcgta ggtctgcagc gcgtagtacc tcacaccgtc
     6781 ctgcgggttg acgtatgagg cttggatcct cccgctgcgc tgagtcttca gcgatcccca
     6841 tccgcgacgt gccaactagg tctcctctcg tcgtgaacaa ggctaccggg ttgcaactcc
     6901 tgtgcaactc tcaggcttca acgcgcttct acgacctgca atttctttcc acttagagga
     6961 tgcagccgag agggggtaaa aacctatctt gaccggccca tatatgtggt cggcagacac
     7021 ccattcttcc aaactagcta cgcgggttcg attcccgtcg cccgctccgc tggtcagagg
     7081 gtgttttcgc cctctggcca tttttctttc caggggtctg caactcttgt gcgactcttc
     7141 tgacctgggc atacgcggtt gcaacgcatc cctgatctgg ctactttcga tgctgacaaa
     7201 cgaatagagc cccccgcctg cgcgaacaga cgaggggcat tcacaccaga ttggagctgg
     7261 tgcagtgaag agaatagacc gggacaaggt tgcaccggga gttgcagcgg tcggaaccct
     7321 cgccgtcggc gggctggcgt tcgccctgtc gttcacggct ctagcg
//
