LOCUS       V015681                 6153 bp    DNA     circular SYN 08-JUL-2019
DEFINITION  Exported.
ACCESSION   V015681
VERSION     V015681
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6153)
  AUTHORS   Bosse JT, Li Y, Leanse LG, Zhou L, Chaudhuri RR, Peters SE, Wang J,
            Maglennon GA, Holden MTG., Maskell DJ, Tucker AW, Wren BW, Rycroft
            AN, Langford PR.
  TITLE     Rationally designed mariner vectors to allow functional genomic
            analysis of Actinobacillus pleuropneumoniae and other bacteria by
            transposon-directed insertion-site sequencing (TraDIS)
  JOURNAL   bioRxivorg (2019) In press
REFERENCE   2  (bases 1 to 6153)
  AUTHORS   Bosse JT.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JUL-2018) Medicine, Imperial College London, Norfolk
            Place, London W2 1PG, UK
REFERENCE   3  (bases 1 to 6153)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxivorg
            (2019) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (17-JUL-2018) Medicine, Imperial College London, Norfolk Place,
            London W2 1PG, UK"
FEATURES             Location/Qualifiers
     source          1..6153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      4..279
                     /label="derived from Actinobacillus pleuropneumoniae sodC"
                     /note="derived from Actinobacillus pleuropneumoniae sodC"
                     /regulatory_class="promoter"
     CDS             289..1335
                     /codon_start=1
                     /transl_table=11
                     /product="transposase"
                     /label="transposase"
                     /note="C9 mutant transposase from Himar1"
                     /protein_id="QDH76837.1"
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDIKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     repeat_region   1437..1465
                     /label="Himar1 IR left"
                     /note="Himar1 IR left"
     misc_feature    1493..1504
                     /label="DNA uptake sequence (DUS) for Neisseria species"
                     /note="DNA uptake sequence (DUS) for Neisseria species"
     misc_feature    1503..1511
                     /note="DNA uptake signal sequence (USS) for Haemophilus
                     influenza"
     misc_feature    1512..1520
                     /note="DNA uptake signal sequence (USS) for Actinobacillus
                     pleuropneumoniae"
     CDS             1690..2337
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     misc_feature    2411..2419
                     /note="DNA uptake signal sequence (USS) for Actinobacillus
                     pleuropneumoniae"
     misc_feature    2419..2430
                     /label="DNA uptake sequence (DUS) for Neisseria species"
                     /note="DNA uptake sequence (DUS) for Neisseria species"
     misc_feature    2431..2439
                     /note="DNA uptake signal sequence (USS) for Haemophilus
                     influenza"
     repeat_region   2442..2470
                     /label="Himar1 IR right"
                     /note="Himar1 IR right"
     oriT            2688..2797
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             2830..3198
                     /label="traJ"
                     /note="oriT-recognizing protein"
     promoter        complement(3278..3296)
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3314..3330)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(3338..3354)
                     /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)."
     promoter        complement(3362..3392)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3407..3428)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3716..4304)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4478..5335)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(5336..5440)
                     /label="AmpR promoter"
     rep_origin      complement(5518..5973)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     6114..6130
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        6137..6153
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgaccg ccaaccgata aaacctacat tttgcatttt atgctcctaa
       61 ttaaatttac taagttgtgt ttgaatttat tttgagaaat agaatctgat aaatacagga
      121 ttttctacta aaatgcaatt agttttagac tatttaaata atattattta aattctttac
      181 tgtagtgtac aatacacaca gtccattaac caaaataaaa ggaggattcc aatcactagt
      241 gaattctgca gtcgacggta ccgcgactag aaaaggagga ttcctcatat ggaaaaaaag
      301 gaatttcgtg ttttgataaa atactgtttt ctgaagggaa aaaatacagt ggaagcaaaa
      361 acttggcttg ataatgagtt tccggactct gccccaggga aatcaacaat aattgattgg
      421 tatgcaaaat tcaagcgtgg tgaaatgagc acggaggacg gtgaacgcag tggacgcccg
      481 aaagaggtgg ttaccgacga aaacatcaaa aaaatccaca aaatgatttt gaatgaccgt
      541 aaaatgaagt tgatcgagat agcagaggcc ttaaagatat caaaggaacg tgttggtcat
      601 atcattcatc aatatttgga tatgcggaag ctctgtgcga aatgggtgcc gcgcgagctc
      661 acatttgacc aaaaacaacg acgtgttgat gattctaagc ggtgtttgca gctgttaact
      721 cgtaatacac ccgagttttt ccgtcgatat gtgacaatgg atgaaacatg gctccatcac
      781 tacactcctg agtccaatcg acagtcggct gagtggacag cgaccggtga accgtctccg
      841 aagcgtggaa agactcaaaa gtccgctggc aaagtaatgg cctctgtttt ttgggatgcg
      901 catggaataa tttttatcga ttatcttgag aagggaaaaa ccatcaacag tgactattat
      961 atggcgttat tggagcgttt gaaggtcgaa atcgcggcaa aacggcccca catgaagaag
     1021 aaaaaagtgt tgttccacca agacaacgca ccgtgccaca agtcattgag aacgatggca
     1081 aaaattcatg aattgggctt cgaattgctt ccccacccgc cgtattctcc agatctggcc
     1141 cccagcgact ttttcttgtt ctcagacatc aaaaggatgc tcgcagggaa aaaatttggc
     1201 tgcaatgaag aggtgatcgc cgaaactgag gcctattttg aggcaaaacc gaaggagtac
     1261 taccaaaatg gtatcaaaaa attggaaggt cgttataatc gttgtatcgc tcttgaaggg
     1321 aactatgttg aataataagc atgctaaggg cgaattccag cacactgggt cgcatgctcc
     1381 cggccgccat ggccgcggga ttattaccct gttatcccta cggccgcact agtgattaac
     1441 aggttggctg ataagtcccc ggtctgcagg cgggcgcact agtgattcgc ggatgccgtc
     1501 tgaagtgcgg tacaagcggt cggcaatagt tacccttaag ttattggtat gactggtttt
     1561 aagcgcaaaa aaagttgctt tttcgtacct attaatgtat cgttttaaaa aaggattgat
     1621 tctaatgaag aaagcagaca agtaagcctc ctaaattcac tttagataaa aatttaggag
     1681 gcatatcaaa tgaactttaa taaaattgat ttagacaatt ggaagagaaa agagatattt
     1741 aatcattatt tgaaccaaca aacgactttt agtataacca cagaaattga tattagtgtt
     1801 ttataccgaa acataaaaca agaaggatat aaattttacc ctgcatttat tttcttagtg
     1861 acaagggtga taaactcaaa tacagctttt agaactggtt acaatagcga cggagagtta
     1921 ggttattggg ataagttaga gccactttat acaatttttg atggtgtatc taaaacattc
     1981 tctggtattt ggactcctgt aaagaatgac ttcaaagagt tttatgattt atacctttct
     2041 gatgtagaga aatataatgg ttcggggaaa ttgtttccca aaacacctat acctgaaaat
     2101 gctttttctc tttctattat tccatggact tcatttactg ggtttaactt aaatatcaat
     2161 aataatagta attaccttct acccattatt acagcaggaa aattcattaa taaaggtaat
     2221 tcaatatatt taccgctatc tttacaggta catcattctg tttgtgatgg ttatcatgca
     2281 ggattgttta tgaactctat tcaggaattg tcagataggc ctaatgactg gcttttataa
     2341 tatgagataa tgccgactgt actttttaca gtcggttttc taatgtcact aacctgcccc
     2401 gttagttgaa accgcttgtt cagacggcat accgcacttc gagaccgggg acttatcagc
     2461 caacctgtta atcccgcggc catgattacc ctgttatccc taatcactag tgcggccgcc
     2521 tgcaggtcga ccaaaacagc agggaagcag cgcttttccg ctgcataacc ctgcttcggg
     2581 gtcattatag cgattttttc ggtatatcca tcctttttcg cacgatatac aggattttgc
     2641 caaagggttc gtgtagactt tccttggtgt atccaacggc gtcagccggg caggataggt
     2701 gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg tcccttattc gcacctggcg
     2761 gtgctcaacg ggaatcctgc tctgcgaggc tggccggcta ccgccggcgt aacagatgag
     2821 ggcaagcgga tggctgatga aaccaagcca accaggaagg gcagcccacc tatcaaggtg
     2881 tactgccttc cagacgaacg aagagcgatt gaggaaaagg cggcggcggc cggcatgagc
     2941 ctgtcggcct acctgctggc cgtcggccag ggctacaaaa tcacgggcgt cgtggactat
     3001 gagcacgtcc gcgagctggc ccgcatcaat ggcgacctgg gccgcctggg cggcctgctg
     3061 aaactctggc tcaccgacga cccgcgcacg gcgcggttcg gtgatgccac gatcctcgcc
     3121 ctgctggcga agatcgaaga gaagcaggac gagcttggca aggtcatgat gggcgtggtc
     3181 cgcccgaggg cagagccatg acttttttag ccgctaaaac ggccgggggg tgcgcgtgat
     3241 tgccaagcac gtccccatgc atgcatagct tgagtattct atagtgtcac ctaaatagct
     3301 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3361 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3421 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3481 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3541 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3601 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3661 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3721 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3781 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3841 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3901 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3961 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4021 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4081 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4141 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4201 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4261 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4321 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4381 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4441 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4501 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4561 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4621 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4681 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4741 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4801 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4861 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4921 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4981 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5041 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5101 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5161 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5221 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5281 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5341 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5401 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5461 cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
     5521 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     5581 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     5641 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     5701 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     5761 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     5821 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     5881 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     5941 ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg
     6001 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     6061 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     6121 gacggccagt gaattgtaat acgactcact ata
//
