LOCUS       V009387                 9406 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009387
VERSION     V009387
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9406)
  AUTHORS   Wilson AC, Perego M, Hoch JA.
  TITLE     New transposon delivery plasmids for insertional mutagenesis in
            Bacillus anthracis
  JOURNAL   J. Microbiol. Methods 71 (3), 332-335 (2007)
   PUBMED   17931726
REFERENCE   2  (bases 1 to 9406)
  AUTHORS   Wilson AC, Perego M, Hoch JA.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-SEP-2007) Department of Molecular and Experimental,
            The Scripps Research Institute, 10550 North Torrey Pines Road, La
            Jolla, CA 92037, USA
REFERENCE   3  (bases 1 to 9406)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9406)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J.
            Microbiol. Methods"; date: "2007"; volume: "71"; issue: "3"; pages:
            "332-335"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (10-SEP-2007) Department of Molecular and Experimental, The Scripps
            Research Institute, 10550 North Torrey Pines Road, La Jolla, CA
            92037, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             171..1217
                     /codon_start=1
                     /product="C9 transposase"
                     /label="C9 transposase"
                     /protein_id="ABV70027.1"
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDLKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     promoter        complement(1280..1298)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1635..1656
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1671..1701
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    1709..1725
                     /label="lac operator"
                     /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)."
     primer_bind     1733..1749
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        1770..1788
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     mobile_element  1801..1827
                     /mobile_element_type="transposon:mariner"
                     /label="ner"
     CDS             1843..1908
                     /label="3xFLAG"
                     /note="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
     protein_bind    1912..1959
                     /label="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        complement(1978..1996)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2483..3071)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             3367..4146
                     /gene="ant1"
                     /label="Spectinomycin 9-adenylyltransferase"
                     /note="Spectinomycin 9-adenylyltransferase from
                     Staphylococcus aureus (strain N315). Accession#: P0A0D1"
     protein_bind    5051..5098
                     /label="FRT"
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     mobile_element  complement(5112..5138)
                     /mobile_element_type="transposon:mariner"
                     /label="ner"
     promoter        complement(5151..5169)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5176..5192)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             5817..6464
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             complement(7080..7778)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label="repA"
                     /protein_id="ABV70030.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNNSNIIQNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            complement(7080..7778)
                     /gene="repA"
                     /label="repA"
     CDS             8390..8401
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
ORIGIN
        1 ccgccagtgt gatggattga cacatagatg gcgtcgctag tattaaatgc atattatttt
       61 tatatagtac caaccttcaa atgattcgtg tcaaaatttg acgtctgtaa gtcaattagt
      121 ttgtgagata gagcgtcttt tgtgaagcaa cttttgttat tgtgaaacat atggaaaaaa
      181 aggaatttcg tgttttgata aaatactgtt ttctgaaggg aaaaaataca gtggaagcaa
      241 aaacttggct tgataatgag tttccggact ctgccccagg gaaatcaaca ataattgatt
      301 ggtatgcaaa attcaagcgt ggtgaaatga gcacggagga cggtgaacgc agtggacgcc
      361 cgaaagaggt ggttaccgac gaaaacatca aaaaaatcca caaaatgatt ttgaatgacc
      421 gtaaaatgaa gttgatcgag atagcagagg ccttaaagat atcaaaggaa cgtgttggtc
      481 atatcattca tcaatatttg gatatgcgga agctctgtgc gaaatgggtg ccgcgcgagc
      541 tcacatttga ccaaaaacaa cgacgtgttg atgattctaa gcggtgtttg cagctgttaa
      601 ctcgtaatac acccgagttt ttccgtcgat atgtgacaat ggatgaaaca tggctccatc
      661 actacactcc tgagtccaat cgacagtcgg ctgagtggac agcgaccggt gaaccgtctc
      721 cgaagcgtgg aaagactcaa aagtccgctg gcaaagtaat ggcctctgtt ttttgggatg
      781 cgcatggaat aatttttatc gattatcttg agaagggaaa aaccatcaac agtgactatt
      841 atatggcgtt attggagcgt ttgaaggtcg aaatcgcggc aaaacggccc cacatgaaga
      901 agaaaaaagt gttgttccac caagacaacg caccgtgcca caagtcattg agaacgatgg
      961 caaaaattca tgaattgggc ttcgaattgc ttccccaccc accgtattct ccagatctgg
     1021 cccccagcga ctttttcttg ttctcagacc tcaaaaggat gctcgcaggg aaaaaatttg
     1081 gctgcaatga agaggtgatc gccgaaactg aggcctattt tgaggcaaaa ccgaaggagt
     1141 actaccaaaa tggtatcaaa aaattggaag gtcgttataa tcgttgtatc gctcttgaag
     1201 ggaactatgt tgaataataa aaacgaattt tgacaaaaaa atgtgttttt ctttgttaga
     1261 ccggaatcct ccaattcgcc ctatagtgag tcgtattacg cgcgggcttt ccccgtcaag
     1321 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     1381 aaaaacttga ttagggtgat ggttcacttc tttcctgcgt tatcccctga ttctgtggat
     1441 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     1501 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     1561 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     1621 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     1681 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     1741 agctatgacc atgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg
     1801 acaggttggc tgataagtcc ccggtctcta gaattcacca tagactacaa agaccatgac
     1861 ggtgattata aagatcatga tatcgattac aaggatgacg atgacaagct tgaagttcct
     1921 attccgaagt tcctattctc tagaaagtat aggaacttcg gcgcgccgct ctagacccct
     1981 atagtgagtc gtattacaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2041 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2101 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     2161 aatgtgcgcg gaaccagatc cccacgatgc gtccggcgta gaggatctga agatcagcag
     2221 ttcaacctgt tgatagtacg atccccctgc attaatgaat cggccaacgc gcggggagag
     2281 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2341 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2401 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2461 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2521 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2581 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2641 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2701 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2761 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2821 gcggactggc agcagccact ggtaacagga ttacgagagc gaggtatgta ggcggtgcta
     2881 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     2941 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3001 aaaccaccgc tggtagcggt ggtttttttg tttcgaagca gcagattacg cgcagaaaaa
     3061 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3121 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3181 tgactctaga ggatcgatct gtataataaa gaataattat taatctgtag acaaattgtg
     3241 aaaggatgta cttaaaccgt aacggtcagc tttattgaac agtaatttaa gtatatgtcc
     3301 aatctagggt aagtaaattg agtatcaata taaactttat atgaacataa tcaacgaggt
     3361 gaaatcatga ccaatttgat taacggaaaa ataccaaatc aagcgattca aacattaaaa
     3421 atcgtaaaag atttatttgg aagttcaata gttggagtat atctatttgg ttcagcagta
     3481 aatggtggtt tacgcattaa cagcgatgta gatggtctag tcgtcgtgaa tcatagttta
     3541 cctcaattaa ctcgaaaaaa actaacagaa agactaatga ctatatcagg aaagattgga
     3601 aatacggatt ctgttagacc acttgaagtt acggttataa ataggagtga agttgtccct
     3661 tggcaatatc ctccaaaaag agaatttata tacggtgagt ggctcagggg tgaatttgag
     3721 aatggacaaa ttcaggaacc aagctatgat cctgatttgg ctattgtttt agcacaagca
     3781 agaaagaata gtatttctct atttggtcct gattcttcaa gtatacttgt ctccgtacct
     3841 ttgacagata ttcgaagagc aattaaggat tctttgccag aactaattga ggggataaaa
     3901 ggtgatgagc gtaatgtaat tttaaccgta gctgcaatgt ggcaaacagt gactactggt
     3961 gaaattacct cgaaagatgt cgctgcagaa tgggctatac ctcttttacc taaagagcat
     4021 gtaactttac tggatatagc tagaaaaggc tatcggggag agtggtatga taagtgggaa
     4081 ggactatatt caaaggtgaa agcactcgtt aagtatatga aaaattctat agaaacttct
     4141 ctcaattagg ctaattttat tcgaataaca ggtgcttact tttaaaacta ctgatttatt
     4201 gataaatatt gaacaatttt tgggaagaat aaagcgtcct cttgtgaaat tagagaacgc
     4261 tttattactt taatttagtg aaacaatttg taactattga aaatagaaag aaattgttcc
     4321 ttgcatagtt tattaatatt agtgacattt gcatgcttca aagcctgtcg gaattggttt
     4381 ttactgaaaa gaacacgata ttcagcgttt acccacttat aaacaaaaga tcgccacgac
     4441 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4501 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4561 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4621 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4681 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4741 gctcaaggcg cggatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4801 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4861 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4921 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4981 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gtacgcgtct
     5041 aggcggccgc gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcga
     5101 accggttcta gagaccgggg acttatcagc caacctgtta gccaattcgc cctatagtga
     5161 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     5221 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     5281 ggcccgcacc gggggatcct ctagagtcga cctgcaggtc gataaaccca gcgaaccatt
     5341 tgaggtgata ggtaagatta taccgaggta tgaaaacgag aattggacct ttacagaatt
     5401 actctatgaa gcgccatatt taaaaagcta ccaagacgaa gaggatgaag aggatgagga
     5461 ggcagattgc cttgaatata ttgacaatac tgataagata atatatcttt tatatagaag
     5521 atagcttgat gggcaatagt tacccttatt atcaagataa gaaagaaaag gatttttcgc
     5581 tacgctcaaa tcctttaaaa aaacacaaaa gaccacattt tttaatgtgg tctttattct
     5641 tcaactaaag cacccattag ttcaacaaac gaaaattgga taaagtggga tatttttaaa
     5701 atatatattt atgttacagt aatattgact tttaaaaaag gattgattct aatgaagaaa
     5761 gcagacaagt aagcctccta aattcacttt agataaaaat ttaggaggca tatcaaatga
     5821 actttaataa aattgattta gacaattgga agagaaaaga gatatttaat cattatttga
     5881 accaacaaac gacttttagt ataaccacag aaattgatat tagtgtttta taccgaaaca
     5941 taaaacaaga aggatataaa ttttaccctg catttatttt cttagtgaca agggtgataa
     6001 actcaaatac agcttttaga actggttaca atagcgacgg agagttaggt tattgggata
     6061 agttagagcc actttataca atttttgatg gtgtatctaa aacattctct ggtatttgga
     6121 ctcctgtaaa gaatgacttc aaagagtttt atgatttata cctttctgat gtagagaaat
     6181 ataatggttc ggggaaattg tttcccaaaa cacctatacc tgaaaatgct ttttctcttt
     6241 ctattattcc atggacttca tttactgggt ttaacttaaa tatcaataat aatagtaatt
     6301 accttctacc cattattaca gcaggaaaat tcattaataa aggtaattca atatatttac
     6361 cgctatcttt acaggtacat cattctgttt gtgatggtta tcatgcagga ttgtttatga
     6421 actctattca ggaattgtca gataggccta atgactggct tttataatat gagataatgc
     6481 cgactgtact ttttacagtc ggttttctaa tgtcactaac ctgccccgtt agttgaagaa
     6541 ggtttttata ttacagctcc agatcatcga attatcgggg aagaacagta tgtcgagcta
     6601 ttttttgact tactggggat caagcctgat tgggagaaaa taaaatatta tattttactg
     6661 gatgaattgt tttagtacct agatttagat gtctaaaaag ctttaactac aagcttttta
     6721 gacatctaat cttttctgaa gtacatccgc aactgtccat actctgatgt tttatatctt
     6781 ttctaaaagt tcgctagata ggggtcccga gcgcctacga ggaatttgta tcgactctag
     6841 aggatcattt tgtttattgc aattgtattg ctattaatcg caacatcaaa ccaaaataaa
     6901 aacccccttc gactttcgtc agggggcttt tatttattca ataatccctc ctctcaataa
     6961 atctattgtt gtacttaatt caacttccat ttctctgtat ctttcaatac gctcttttaa
     7021 gtccttaatt tcttttttta attcctcatt ttcagcaaat aactcttttt ctttgtttgt
     7081 cattttattt cccccgtttc agcatcaaga acctttgcat aacttgctct atatccacac
     7141 tgataattgc cctcaaacca taatctaaag gcgctagagt ttgttgaaac aatatctttt
     7201 acatcattcg tatttaaaat tccaaactcc gctcccctaa ggcgaataaa agccattaaa
     7261 tcttttgtat ttaccaaatt atagtcatcc actatatcta aaagtaaatt cttcaattct
     7321 cttttttggc tttcatcaag tgttatatag cggtcaatat caaaatcatt aatgttcaaa
     7381 atatcttttt tgtcgtatat atgtttattc ttagcaatag cgtcctttga ttcatgagtc
     7441 aaatattcat atgaaccttt gatataatca agtatctcaa catgagcaac tgaactattc
     7501 cccaattttc gcttaatctt gttcctaacg ctttctattg ttacaggatt tcgtgcaata
     7561 tatataacgt gatagtgtgg ttttttatag tgctttccat tttgtataat attactatta
     7621 ttccatgtat ctttatcttt tttttcgtcc atatcgtgta aaggactgac agccatagat
     7681 acgcccaaac tctctaattt ttctttccaa tcattaggaa ttgagtcagg atataataaa
     7741 aatccaaaat ttctagcttt agtattttta atagccatga tataattacc ttatcaaaaa
     7801 caagtagcga aaactcgtat ccttctaaaa acgcgagctt tcgcttattt tttttattct
     7861 gattcctttc ttgcatattc ttctatagct aacgccgcaa ccgcagattt tgaaaaacct
     7921 ttttgtttcg ccatatctgt taatttttta tcttgctctt ttgtcagaga aatcataact
     7981 ctttttttcg attctgaaat caccatttaa aaaactccaa tcaaataatt ttataaagtt
     8041 agtgtatcac tttgtaatca taaaaacaac aataaagcta cttaaatata gatttataaa
     8101 aaacgttggc gaaaacgttg gcgattcgtt ggcgattgaa aaacccctta aacccttgag
     8161 ccagttggga tagagcgttt ttggcacaaa aattggcact cggcacttaa tggggggtcg
     8221 tagtacggaa gcaaaattcg cttcctttcc ccccattttt ttccaaattc caaatttttt
     8281 tcaaaaattt tccagcgcta ccgctcggca aaattgcaag caatttttaa aatcaaaccc
     8341 atgagggaat ttcattccct catactccct tgagcctcct ccaaccgaaa tagaagggcg
     8401 ctgcgcttat tatttcattc agtcatcggc tttcataatc taacagacaa catcttcgct
     8461 gcaaagccac gctacgctca agggctttta cgctacgata acgcctgttt taacgattat
     8521 gccgataact aaacgaaata aacgctaaaa cgtctcagaa acgattttga gacgttttaa
     8581 taaaaaatcg gataataaat atatataaac gtatatagat ttcataaagt ctaacacact
     8641 agacttattt acttcgtaat taagtcgtta aaccgtgtgc tctacgacca aaactataaa
     8701 acctttaaga actttctttt tttacaagaa aaaagaaatt agataaatct ctcatatctt
     8761 ttattcaata atcgcatccg attgcagtat aaatttaacg atccccatcg gaggaccgaa
     8821 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     8881 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     8941 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     9001 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     9061 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     9121 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     9181 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     9241 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     9301 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     9361 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gagcgg
//
