LOCUS       V015351                 6050 bp    DNA     circular SYN 08-JUL-2019
DEFINITION  Exported.
ACCESSION   V015351
VERSION     V015351
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6050)
  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 6050)
  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 6050)
  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..6050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    35..56
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        71..101
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    109..125
                     /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     133..149
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             186..1232
                     /codon_start=1
                     /transl_table=11
                     /product="transposase"
                     /label="transposase"
                     /note="C9 mutant transposase from Himar1"
                     /protein_id="QDH76841.1"
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDIKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     repeat_region   1334..1362
                     /label="Himar1 IR left"
                     /note="Himar1 IR left"
     misc_feature    1390..1401
                     /label="DNA uptake sequence (DUS) for Neisseria species"
                     /note="DNA uptake sequence (DUS) for Neisseria species"
     misc_feature    1400..1408
                     /note="DNA uptake signal sequence (USS) for Haemophilus
                     influenza"
     misc_feature    1409..1417
                     /note="DNA uptake signal sequence (USS) for Actinobacillus
                     pleuropneumoniae"
     CDS             1587..2234
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     misc_feature    2308..2316
                     /note="DNA uptake signal sequence (USS) for Actinobacillus
                     pleuropneumoniae"
     misc_feature    2316..2327
                     /label="DNA uptake sequence (DUS) for Neisseria species"
                     /note="DNA uptake sequence (DUS) for Neisseria species"
     misc_feature    2328..2336
                     /note="DNA uptake signal sequence (USS) for Haemophilus
                     influenza"
     repeat_region   2339..2367
                     /label="Himar1 IR right"
                     /note="Himar1 IR right"
     oriT            2585..2694
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             2727..3095
                     /label="traJ"
                     /note="oriT-recognizing protein"
     promoter        complement(3175..3193)
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3211..3227)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(3235..3251)
                     /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(3259..3289)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3304..3325)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3613..4201)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4375..5232)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(5233..5337)
                     /label="AmpR promoter"
     rep_origin      complement(5415..5870)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     6011..6027
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        6034..6050
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacgt gagcgcaacg caattaatgt gagttagctc actcattagg
       61 caccccaggc tttacacttt atgctcccgg ctcgtatgtt gtgtggaatt gtgagcggat
      121 aacaatttca cacaggaaac agctatgacc atgattacgc cgactagaaa aggaggattc
      181 ctcatatgga aaaaaaggaa tttcgtgttt tgataaaata ctgttttctg aagggaaaaa
      241 atacagtgga agcaaaaact tggcttgata atgagtttcc ggactctgcc ccagggaaat
      301 caacaataat tgattggtat gcaaaattca agcgtggtga aatgagcacg gaggacggtg
      361 aacgcagtgg acgcccgaaa gaggtggtta ccgacgaaaa catcaaaaaa atccacaaaa
      421 tgattttgaa tgaccgtaaa atgaagttga tcgagatagc agaggcctta aagatatcaa
      481 aggaacgtgt tggtcatatc attcatcaat atttggatat gcggaagctc tgtgcgaaat
      541 gggtgccgcg cgagctcaca tttgaccaaa aacaacgacg tgttgatgat tctaagcggt
      601 gtttgcagct gttaactcgt aatacacccg agtttttccg tcgatatgtg acaatggatg
      661 aaacatggct ccatcactac actcctgagt ccaatcgaca gtcggctgag tggacagcga
      721 ccggtgaacc gtctccgaag cgtggaaaga ctcaaaagtc cgctggcaaa gtaatggcct
      781 ctgttttttg ggatgcgcat ggaataattt ttatcgatta tcttgagaag ggaaaaacca
      841 tcaacagtga ctattatatg gcgttattgg agcgtttgaa ggtcgaaatc gcggcaaaac
      901 ggccccacat gaagaagaaa aaagtgttgt tccaccaaga caacgcaccg tgccacaagt
      961 cattgagaac gatggcaaaa attcatgaat tgggcttcga attgcttccc cacccgccgt
     1021 attctccaga tctggccccc agcgactttt tcttgttctc agacatcaaa aggatgctcg
     1081 cagggaaaaa atttggctgc aatgaagagg tgatcgccga aactgaggcc tattttgagg
     1141 caaaaccgaa ggagtactac caaaatggta tcaaaaaatt ggaaggtcgt tataatcgtt
     1201 gtatcgctct tgaagggaac tatgttgaat aataagcatg ctaagggcga attccagcac
     1261 actgggtcgc atgctcccgg ccgccatggc cgcgggatta ttaccctgtt atccctacgg
     1321 ccgcactagt gattaacagg ttggctgata agtccccggt ctgcaggcgg gcgcactagt
     1381 gattcgcgga tgccgtctga agtgcggtac aagcggtcgg caatagttac ccttaagtta
     1441 ttggtatgac tggttttaag cgcaaaaaaa gttgcttttt cgtacctatt aatgtatcgt
     1501 tttaaaaaag gattgattct aatgaagaaa gcagacaagt aagcctccta aattcacttt
     1561 agataaaaat ttaggaggca tatcaaatga actttaataa aattgattta gacaattgga
     1621 agagaaaaga gatatttaat cattatttga accaacaaac gacttttagt ataaccacag
     1681 aaattgatat tagtgtttta taccgaaaca taaaacaaga aggatataaa ttttaccctg
     1741 catttatttt cttagtgaca agggtgataa actcaaatac agcttttaga actggttaca
     1801 atagcgacgg agagttaggt tattgggata agttagagcc actttataca atttttgatg
     1861 gtgtatctaa aacattctct ggtatttgga ctcctgtaaa gaatgacttc aaagagtttt
     1921 atgatttata cctttctgat gtagagaaat ataatggttc ggggaaattg tttcccaaaa
     1981 cacctatacc tgaaaatgct ttttctcttt ctattattcc atggacttca tttactgggt
     2041 ttaacttaaa tatcaataat aatagtaatt accttctacc cattattaca gcaggaaaat
     2101 tcattaataa aggtaattca atatatttac cgctatcttt acaggtacat cattctgttt
     2161 gtgatggtta tcatgcagga ttgtttatga actctattca ggaattgtca gataggccta
     2221 atgactggct tttataatat gagataatgc cgactgtact ttttacagtc ggttttctaa
     2281 tgtcactaac ctgccccgtt agttgaaacc gcttgttcag acggcatacc gcacttcgag
     2341 accggggact tatcagccaa cctgttaatc ccgcggccat gattaccctg ttatccctaa
     2401 tcactagtgc ggccgcctgc aggtcgacca aaacagcagg gaagcagcgc ttttccgctg
     2461 cataaccctg cttcggggtc attatagcga ttttttcggt atatccatcc tttttcgcac
     2521 gatatacagg attttgccaa agggttcgtg tagactttcc ttggtgtatc caacggcgtc
     2581 agccgggcag gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc
     2641 cttattcgca cctggcggtg ctcaacggga atcctgctct gcgaggctgg ccggctaccg
     2701 ccggcgtaac agatgagggc aagcggatgg ctgatgaaac caagccaacc aggaagggca
     2761 gcccacctat caaggtgtac tgccttccag acgaacgaag agcgattgag gaaaaggcgg
     2821 cggcggccgg catgagcctg tcggcctacc tgctggccgt cggccagggc tacaaaatca
     2881 cgggcgtcgt ggactatgag cacgtccgcg agctggcccg catcaatggc gacctgggcc
     2941 gcctgggcgg cctgctgaaa ctctggctca ccgacgaccc gcgcacggcg cggttcggtg
     3001 atgccacgat cctcgccctg ctggcgaaga tcgaagagaa gcaggacgag cttggcaagg
     3061 tcatgatggg cgtggtccgc ccgagggcag agccatgact tttttagccg ctaaaacggc
     3121 cggggggtgc gcgtgattgc caagcacgtc cccatgcatg catagcttga gtattctata
     3181 gtgtcaccta aatagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3241 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3301 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3361 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3421 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3481 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3541 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3601 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3661 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3721 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3781 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3841 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3901 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3961 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4021 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4081 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4141 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4201 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4261 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4321 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4381 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4441 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4501 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4561 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4621 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4681 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4741 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4801 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4861 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4921 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4981 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5041 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5101 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5161 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5221 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5281 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5341 atttccccga aaagtgccac ctgatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     5401 aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     5461 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     5521 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     5581 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     5641 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     5701 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     5761 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     5821 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca
     5881 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     5941 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     6001 tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata
//
