LOCUS       40924_15180        5664 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDS1028, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5664)
  AUTHORS   Monson RE, Drew AM, Smith D, Matilla M, Roberts K, Richardson E, 
            Williamson N, Ramsay J, Welch M, Salmond GPC.
  TITLE     A plasmid-transposon hybrid mutagenesis system effective in a broad 
            range of Enterobacteria
  JOURNAL   Front Microbiol (2016) In press
REFERENCE   2  (bases 1 to 5664)
  AUTHORS   Monson RE, Smith D, Matilla M, Roberts K, Richardson E, Drew A, 
            Williamson N, Ramsay J, Welch M, Salmond GPC.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-NOV-2015) Biochemistry, University of Cambridge, 
            Tennis Court Road, Cambridge CB2 1QW, United Kingdom
REFERENCE   3  (bases 1 to 5664)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5664)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front 
            Microbiol (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-NOV-2015) Biochemistry, University of Cambridge, Tennis Court 
            Road, Cambridge CB2 1QW, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(132..1319)
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     promoter        complement(1367..1395)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             1773..3200
                     /codon_start=1
                     /label=Tn5 transposase
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAAQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     misc_feature    3212..3230
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     rep_origin      complement(3269..3657)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     promoter        3974..4076
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             4077..4733
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     misc_feature    complement(5121..5139)
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     oriT            complement(5378..5487)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
ORIGIN
        1 gaattcctgc agcccggggg atccactagt tctagggttg gtttgcgcat tcacagttct
       61 ccgcaagaat tgattggctc caattcttgg agtggtgaat ccgttagcga ggtgccgccg
      121 gcttccattc aggtcgaggt ggcccggctc catgcaccgc gacgcaacgc ggggaggcag
      181 acaaggtata gggcggcgcc tacaatccat gccaacccgt tccatgtgct cgccgaggcg
      241 gcataaatcg ccgtgacgat cagcggtcca gtgatcgaag ttaggctggt aagagccgcg
      301 agcgatcctt gaagctgtcc ctgatggtcg tcatctacct gcctggacag catggcctgc
      361 aacgcgggca tcccgatgcc gccggaagcg agaagaatca taatggggaa ggccatccag
      421 cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt cggccgccat gccggcgata
      481 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
      541 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
      601 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
      661 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
      721 actgggttga aggctctcaa gggcatcggt cgacgctctc ccttatgcga ctcctgcatt
      781 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
      841 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
      901 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
      961 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     1021 tagaggatcc acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt
     1081 agcgaagcga gcaggactgg gcggcggcca aagcggtcgg acagtgctcc gagaacgggt
     1141 gcgcatagaa attgcatcaa cgcatatagc gctagcagca cgccatagtg actggcgatg
     1201 ctgtcggaat ggacgatatc ccgcaagagg cccggcagta ccggcataac caagcctatg
     1261 cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata
     1321 cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg
     1381 atgataagct gtcaaacatg agaattacaa cttatatcgt atggggctga cttcaggtgc
     1441 tacatttgaa gagataaatt gcactgaaat ctagaatgat tctccgccag catggcttcg
     1501 gccagtgcgt cgagcagcgc ccgcttgttc ctgaagtgcc agtaaagcgc cggctgctga
     1561 acccccaacc gttccgccag tttgcgtgtc gtcagaccgt ctacgccgac ctcgttcaac
     1621 aggtctaggg cggcacggat cactgtattc ggctgcaact ttgtcatgct tgacacttta
     1681 tcactgataa acataatatg tccaccaact tatcagtgat aaagaatccg cgcgttcaat
     1741 cggaccagcg gaggctggtc cggaggccac atatgataac ttctgctctt catcgtgcgg
     1801 ccgactgggc taaatctgtg ttctcttcgg cggcgctggg tgatcctcgc cgtactgccc
     1861 gcttggttaa cgtcgccgcc caattggcaa aatattctgg taaatcaata accatctcat
     1921 cagagggtag taaagccgcc caggaaggcg cttaccgatt tatccgcaat cccaacgttt
     1981 ctgccgaggc gatcagaaag gctggcgcca tgcaaacagt caagttggct caggagtttc
     2041 ccgaactgct ggccattgag gacaccacct ctttgagtta tcgccaccag gtcgccgaag
     2101 agcttggcaa gctgggctct attcaggata aatcccgcgg atggtgggtt cactccgttc
     2161 tcttgctcga ggccaccaca ttccgcaccg taggattact gcatcaggag tggtggatgc
     2221 gcccggatga ccctgccgat gcggatgaaa aggagagtgg caaatggctg gcagcggccg
     2281 caactagccg gttacgcatg ggcagcatga tgagcaacgt gattgcggtc tgtgaccgcg
     2341 aagccgatat tcatgcttat ctgcaggaca aactggcgca taacgagcgc ttcgtggtgc
     2401 gctccaagca cccacgcaag gacgtagagt ctgggttgta tctgtacgac catctgaaga
     2461 accaaccgga gttgggtggc tatcagatca gcattccgca aaagggcgtg gtggataaac
     2521 gcggtaaacg taaaaatcga ccagcccgca aggcgagctt gagcctgcgc agtgggcgca
     2581 tcacgctaaa acaggggaat atcacgctca acgcggtgct ggccgaggag attaacccgc
     2641 ccaagggtga gaccccgttg aaatggttgt tgctgaccag cgaaccggtc gagtcgctag
     2701 cccaagcctt gcgcgtcatc gacatttata cccatcgctg gcggatcgag gagttccata
     2761 aggcatggaa aaccggagca ggagccgaga ggcaacgcat ggaggagccg gataatctgg
     2821 agcggatggt ctcgatcctc tcgtttgttg cggtcaggct gttacagctc agagaaagct
     2881 tcacgccgcc gcaagcactc agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa
     2941 gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc tatctggaca
     3001 agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac atggcgatag
     3061 ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg ggcgccctct
     3121 gggaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc aaggatctga
     3181 tggcgcaggg gatcaagatc tgatccgtcg actgtctctt atacacatct caaccatcat
     3241 cgatgaattc gagctcggta cccccccatg tcagccgtta agtgttcctg tgtcactcaa
     3301 aattgctttg agaggctcta agggcttctc agtgcgttac atccctggct tgttgtccac
     3361 aaccgttaaa ccttaaaagc tttaaaagcc ttatatattc ttttttttct tataaaactt
     3421 aaaaccttag aggctattta agttgctgat ttatattaat tttattgttc aaacatgaga
     3481 gcttagtacg tgaaacatga gagcttagta cgttagccat gagagcttag tacgttagcc
     3541 atgagggttt agttcgttaa acatgagagc ttagtacgtt aaacatgaga gcttagtacg
     3601 tgaaacatga gagcttagta cgtactatca acaggttgaa ctgctgatct tcagatcctc
     3661 tacgccggac gcatcgtggc cggggttcga aatcgatgat acactccgct agcgctgatg
     3721 tccggcggtg cttttgccgt tacgcaccac cccgtcagta gctgaacagg agggacagct
     3781 gatagaaaca gaagccactg gagcacctca aaaacaccat catacactaa atcagtaagt
     3841 tggcagcatc acccgacgca ctttgcgccg aataaatacc tgtgacggaa gatcacttcg
     3901 cagaataaat aaatcctggt gtccctgttg ataccgggaa gccctgggcc aacttttggc
     3961 gaaaatgaga cgttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat
     4021 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg
     4081 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
     4141 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
     4201 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     4261 ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc
     4321 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
     4381 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
     4441 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
     4501 tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
     4561 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca
     4621 aggtgctgat gccgctggcg attcaggttc atcatgccgt ctgtgatggc ttccatgtcg
     4681 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt
     4741 tttaaggcag ttattggtgc ccttaaacgc ctggtgctac gcctgaataa gtgataataa
     4801 gcggatgaat ggcagaaatt cgataccgtc gcttaattaa ccgagagctt ggcacccagc
     4861 ctgcgcgagc aggggaattg atccggtgga tgaccttttg aatgaccttt aatagattat
     4921 attactaatt aattggggac cctagaggtc ccctttttta ttttaaaaat tttttcacaa
     4981 aacggtttac aagcataaag cttgctcaat caatcaccgg atccccggga attctcatgt
     5041 ttgacagctt atcatcgata agcttctgcc tcgcgcgata aatctagagt cgacctgcag
     5101 gcatgcaagc ttcagggttg agatgtgtat aagagacagt cgacctgcag gatcgatggc
     5161 gcgccgacgt gctcatagtc cacgacgccc gtgattttgt agccctggcc gacggccagc
     5221 aggtaggccg acaggctcat gccggccgcc gccgcctttt cctcaatcgc tcttcgttcg
     5281 tctggaaggc agtacacctt gataggtggg ctgcccttcc tggttggctt ggtttcatca
     5341 gccatccgct tgccctcatc tgttacgccg gcggtagccg gccagcctcg cagagcagga
     5401 ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg
     5461 ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct
     5521 acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa
     5581 atcgctataa tgaccccgaa gcagggttat gcagcggaaa agcgctgctt ccctgctgtt
     5641 ttgtggcgcg ccatcgatcc gccg
//
