LOCUS       V002265                 8952 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002265
VERSION     V002265
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8952)
  AUTHORS   Li R, Wang G, Shen B, Wang R, Song Y, Li S, Jiang J.
  TITLE     Random transposon vectors pUTTns for the markerless integration of
            exogenous genes into gram-negative eubacteria chromosomes
  JOURNAL   J. Microbiol. Methods 79 (2), 220-226 (2009)
   PUBMED   19778558
REFERENCE   2  (bases 1 to 8952)
  AUTHORS   Li R, Jiang J, Li S.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-JUL-2009) College of Life Science, Nanjing
            Agricultural University, 6# Tongwei Road, Nanjing, Jiangsu 210095,
            China
REFERENCE   3  (bases 1 to 8952)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8952)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J.
            Microbiol. Methods"; date: "2009"; volume: "79"; issue: "2"; pages:
            "220-226"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (09-JUL-2009) College of Life Science, Nanjing Agricultural
            University, 6# Tongwei Road, Nanjing, Jiangsu 210095, China"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(314..1171)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(1172..1276)
                     /label="AmpR promoter"
     CDS             complement(1326..2753)
                     /label="Tn5 transposase"
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"
     misc_feature    2871..2889
                     /label="Tn5 IE end"
                     /note="Tn5 IE end"
     CDS             complement(3256..4674)
                     /label="SacB"
                     /note="secreted levansucrase that renders bacterial growth
                     sensitive to sucrose"
     CDS             5333..5980
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     misc_feature    6123..6554
                     /label="similar to sacB"
                     /note="similar to sacB"
     misc_feature    6555..6584
                     /label="MCS"
                     /note="MCS"
     misc_feature    6585..6603
                     /label="Tn5 OE end"
                     /note="Tn5 OE end"
     protein_bind    complement(6615..6631)
                     /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)."
     rep_origin      complement(6666..7054)
                     /direction=LEFT
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(7675..8043)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(8076..8185)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
ORIGIN
        1 gccccgtggc cgggggactg ttgggcgcca tctccttgct gcctcgcgcg tttcggtgat
       61 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
      121 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
      181 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
      241 cagagcagat tgtactgaga gtgcaccata aatcaatcta aagtatatat gagtaaactt
      301 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
      361 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
      421 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
      481 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
      541 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
      601 gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta
      661 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
      721 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
      781 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
      841 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
      901 gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt
      961 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     1021 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     1081 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     1141 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     1201 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     1261 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgcagatctg caggtcgacc
     1321 gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta
     1381 ctttgcaggg cttcccaacc ttcccagagg gcgccccagc tggcaattcc ggttcgcttg
     1441 ctgtccataa aaccgcccag tctagctatc gccatgtaag cccactgcaa gctacctgct
     1501 ttctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg
     1561 gtcagcaccg tttctgcgga ctggctttct acgtgttccg cttcctttag cagcccttgc
     1621 gccctgagtg cttgcggcag cgtgaagctt tctctgagct gtaacagcct gaccgcaaca
     1681 aacgagagga tcgagaccat ccgctccaga ttatccggct cctccatgcg ttgcctctcg
     1741 gctcctgctc cggttttcca tgccttatgg aactcctcga tccgccagcg atgggtataa
     1801 atgtcgatga cgcgcaaggc ttgggctagc gactcgaccg gttcgctggt cagcaacaac
     1861 catttcaacg gggtctcacc cttgggcggg ttaatctcct cggccagcac cgcgttgagc
     1921 gtgatattcc cctgttttag cgtgatgcgc ccactgcgca ggctcaagct cgccttgcgg
     1981 gctggtcgat ttttacgttt accgcgttta tccaccacgc ccttttgcgg aatgctgatc
     2041 tgatagccac ccaactccgg ttggttcttc agatggtcgt acagatacaa cccagactct
     2101 acgtccttgc gtgggtgctt ggagcgcacc acgaagcgct cgttatgcgc cagtttgtcc
     2161 tgcagataag catgaatatc ggcttcgcgg tcacagaccg caatcacgtt gctcatcatg
     2221 ctgcccatgc gtaaccggct agttgcggcg gctgccagcc atttgccact ctccttttca
     2281 tccgcatcgg cagggtcatc cgggcgcatc caccactcct gatgcagtaa tcctacggtg
     2341 cggaatgtgg tggcctcgag caagagaacg gagtgaaccc accatccgcg ggatttatcc
     2401 tgaatagagc ccagcttgcc aagctcttcg gcgacctggt ggcgataact caaagaggtg
     2461 gtgtcctcaa tggccagcag ttcgggaaac tcctgagcca acttgactgt ttgcatggcg
     2521 ccagcctttc tgatcgcctc ggcagaaacg ttgggattgc ggataaatcg gtaagcgcct
     2581 tcctgcatgg cttcactacc ctctgatgag atggttattg atttaccaga atattttgcc
     2641 aattgggcgg cgacgttaac caagcgggca gtacggcgag gatcacccag cgccgccgaa
     2701 gagaacacag atttagccca gtcggccgca cgatgaagag cagaagttat catgaacgtt
     2761 accatgttag gaggtcacat ggaagatcag atcctggaaa acgggaaagg ttccgttcag
     2821 gacgctactt gtgtacggga tcggtcgacg gatcccaagc ttcttctaga ctgtctcttg
     2881 atcagatctg gccgcctagg ccgaattcga gctccttccc aaccttacca gagggcgccc
     2941 cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg
     3001 taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt gtccagatag
     3061 cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct ttctacgtgt
     3121 tccgcttcct ttagcagccc ttgcgccctg agtgcttgcg gcagcgtgaa gctcgaggta
     3181 tgggattcac ctttatgttg ataagaaata aaagaaaatg ccaataggat atcggcattt
     3241 tcttttgcgt ttttatttgt taactgttaa ttgtccttgt tcaaggatgc tgtctttgac
     3301 aacagatgtt ttcttgcctt tgatgttcag caggaagctt ggcgcaaacg ttgattgttt
     3361 gtctgcgtag aatcctctgt ttgtcatata gcttgtaatc acgacattgt ttcctttcgc
     3421 ttgaggtaca gcgaagtgtg agtaagtaaa ggttacatcg ttaggatcaa gatccatttt
     3481 taacacaagg ccagttttgt tcagcggctt gtatgggcca gttaaagaat tagaaacata
     3541 accaagcatg taaatatcgt tagacgtaat gccgtcaatc gtcatttttg atccgcggga
     3601 gtcagtgaac aggtaccatt tgccgttcat tttaaagacg ttcgcgcgtt caatttcatc
     3661 tgttactgtg ttagatgcaa tcagcggttt catcactttt ttcagtgtgt aatcatcgtt
     3721 tagctcaatc ataccgagag cgccgtttgc taactcagcc gtgcgttttt tatcgctttg
     3781 cagaagtttt tgactttctt gacggaagaa tgatgtgctt ttgccatagt atgctttgtt
     3841 aaataaagat tcttcgcctt ggtagccatc ttcagttcca gtgtttgctt caaatactaa
     3901 gtatttgtgg cctttatctt ctacgtagtg aggatctctc agcgtatggt tgtcgcctga
     3961 gctgtagttg ccttcatcga tgaactgctg tacattttga tacgtttttc cgtcaccgtc
     4021 aaagattgat ttataatcct ctacaccgtt gatgttcaaa gagctgtctg atgctgatac
     4081 gttaacttgt gcagttgtca gtgtttgttt gccgtaatgt ttaccggaga aatcagtgta
     4141 gaataaacgg atttttccgt cagatgtaaa tgtggctgaa cctgaccatt cttgtgtttg
     4201 gtcttttagg atagaatcat ttgcatcgaa tttgtcgctg tctttaaaga cgcggccagc
     4261 gtttttccag ctgtcaatag aagtttcgcc gactttttga tagaacatgt aaatcgatgt
     4321 gtcatccgca tttttaggat ctccggctaa tgcaaagacg atgtggtagc cgtgatagtt
     4381 tgcgacagtg ccgtcagcgt tttgtaatgg ccagctgtcc caaacgtcca ggccttttgc
     4441 agaagagata tttttaattg tggacgaatc gaattcagga acttgatatt tttcattttt
     4501 ttgctgttca gggatttgca gcatatcatg gcgtgtaata tgggaaatgc cgtatgtttc
     4561 cttatatggc ttttggttcg tttctttcgc aaacgcttga gttgcgcctc ctgccagcag
     4621 tgcggtagta aaggttaata ctgttgcttg ttttgcaaac tttttgatgt tcatcgttca
     4681 tgtctccttt tttatgtact gtgttagcgg tctgcttctt ccagccctcc tgtttgaaga
     4741 tggcaagtta gttacgcaca ataaaaaaag acctaaaata tgtaaggggt gacgccaaag
     4801 tatacacttt gccctttaca cattttaggt cttgcctgct ttatcagtaa caaacccgcg
     4861 cgatttactt ttcgacctca ttctattaga ctctcgtttg gattgcaact ggtctatttt
     4921 cctcttttgt ttgatagaaa atcataaaag gatttgcaga ctacgggcct aaagaactag
     4981 agctcggtac ccggggatcc ggctgtaatc cgggcagcgc aacggaacat tcatcagtgt
     5041 aaaaatggaa tcaataaagc cctgcgcagc gcgcagggtc agcctgaata cgcgtactag
     5101 taccttgatg acacagaaga aggcgatttg attcatacag atgatgacga aaaagccgat
     5161 gaagatggat tttctattat tgcaatgtgg aattgggaac ggaaaaatta ttttattaaa
     5221 gagtagttca acaaacggga ttgactttta aaaaaggatt gattctaatg aagaaagcag
     5281 acaagtaagc ctcctaaatt cactttagat aaaaatttag gaggcatatc aaatgaactt
     5341 taataaaatt gatttagaca attggaagag aaaagagata tttaatcatt atttgaacca
     5401 acaaacgact tttagtataa ccacagaaat tgatattagt gttttatacc gaaacataaa
     5461 acaagaagga tataaatttt accctgcatt tattttctta gtgacaaggg tgataaactc
     5521 aaatacagct tttagaactg gttacaatag cgacggagag ttaggttatt gggataagtt
     5581 agagccactt tatacaattt ttgatggtgt atctaaaaca ttctctggta tttggactcc
     5641 tgtaaagaat gacttcaaag agttttatga tttatacctt tctgatgtag agaaatataa
     5701 tggttcgggg aaattgtttc ccaaaacacc tatacctgaa aatgcttttt ctctttctat
     5761 tattccatgg acttcattta ctgggtttaa cttaaatatc aataataata gtaattacct
     5821 tctacccatt attacagcag gaaaattcat taataaaggt aattcaatat atttaccgct
     5881 atctttacag gtacatcatt ctgtttgtga tggttatcat gcaggattgt ttatgaactc
     5941 tattcaggaa ttgtcagata ggcctaatga ctggctttta taagacgtca cgcgtattca
     6001 ggctgaccct gcgcgctgcg cagggcttta ttgattccat ttttacactg atgaatgttc
     6061 cgttgcgctg cccggattac agccggatcc tctagagtcg acctgcaggc atgcgacaac
     6121 agatgttttc ttgcctttga tgttcagcag gaagcttggc gcaaacgttg attgtttgtc
     6181 tgcgtagaat cctctgtttg tcatatagct tgtaatcacg acattgtttc ctttcgcttg
     6241 aggtacagcg aagtgtgagt aagtaaaggt tacatcgtta ggatcaagat ccatttttaa
     6301 cacaaggcca gttttgttca gcggcttgta tgggccagtt aaagaattag aaacataacc
     6361 aagcatgtaa atatcgttag acgtaatgcc gtcaatcgtc atttttgatc cgcgggagtc
     6421 agtgaacagg taccatttgc cgttcatttt aaagacgttc gcgcgttcaa tttcatctgt
     6481 tactgtgtta gatgcaatca gcggtttcat cacttttttc agtgtgtaat catcgtttag
     6541 ctcaatcata ccgagggccc atggcatatg tccggagcgg ccgcacttgt gtataagagt
     6601 cagaattcca caaattgtta tccgctcaca attccacatg tggaattcca catgtggaat
     6661 tcccatgtca gccgttaagt gttcctgtgt cactcaaaat tgctttgaga ggctctaagg
     6721 gcttctcagt gcgttacatc cctggcttgt tgtccacaac cgttaaacct taaaagcttt
     6781 aaaagcctta tatattcttt tttttcttat aaaacttaaa accttagagg ctatttaagt
     6841 tgctgattta tattaatttt attgttcaaa catgagagct tagtacgtga aacatgagag
     6901 cttagtacgt tagccatgag agcttagtac gttagccatg agggtttagt tcgttaaaca
     6961 tgagagctta gtacgttaaa catgagagct tagtacgtga aacatgagag cttagtacgt
     7021 actatcaaca ggttgaactg ctgatcttca gatcctctac gccggacgca tcgtggccgg
     7081 atccagccga ccaggctttc cacgcccgcg tgccgctcca tgtcgttcgc gcggttctcg
     7141 gaaacgcgct gccgcgtttc gtgattgtca cgctcaagcc cgtagtcccg ttcgagcgtc
     7201 gcgcagaggt cagcgagggc gcggtaggcc cgatacggct catggatggt gtttcgggtc
     7261 gggtgaatct tgttgatggc gatatggatg tgcaggttgt cggtgtcgtg atgcacggca
     7321 ctgacgcgct gatgctcggc gaagccaagc ccagcgcaga tgcggtcctc aatcgcgcgc
     7381 aacgtctccg cgtcgggctt ctctcccgcg cggaagctaa ccagcaggtg ataggtcttg
     7441 tcggcctcgg aacgggtgtt gccgtgctgg gtcgccatca cctcggccat gacagcgggc
     7501 agggtgtttg cctcgcagtt cgtgacgcgc acgtgaccca ggcgctcggt cttgccttgc
     7561 tcgtcggtga tgtacttcac cagctccgcg aagtcgctct tcttgatgga gcgcatgggg
     7621 acgtgcttgg caatcacgcg caccccccgg ccgttttagc ggctaaaaaa gtcatggctc
     7681 tgccctcggg cggaccacgc ccatcatgac cttgccaagc tcgtcctgct tctcttcgat
     7741 cttcgccagc agggcgagga tcgtggcatc accgaaccgc gccgtgcgcg ggtcgtcggt
     7801 gagccagagt ttcagcaggc cgcccaggcg gcccaggtcg ccattgatgc gggccagctc
     7861 gcggacgtgc tcatagtcca cgacgcccgt gattttgtag ccctggccga cggccagcag
     7921 gtaggccgac aggctcatgc cggccgccgc cgccttttcc tcaatcgctc ttcgttcgtc
     7981 tggaaggcag tacaccttga taggtgggct gcccttcctg gttggcttgg tttcatcagc
     8041 catccgcttg ccctcatctg ttacgccggc ggtagccggc cagcctcgca gagcaggatt
     8101 cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc cgctcgcggg
     8161 tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag gaaagtctac
     8221 acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat
     8281 cgctataatg accccgaagc agggttatgc agcggaaaag cgctgcttcc ctgctgtttt
     8341 gtggaatatc taccgactgg aaacaggcaa atgcaggaaa ttactgaact gaggggacag
     8401 gcgagagacg atgccaaaga gctacaccga cgagctggcc gagtgggttg aatcccgcgc
     8461 ggccaagaag cgccggcgtg atgaggctgc ggttgcgttc ctggcggtga gggcggatgt
     8521 cgaggcggcg ttagcgtccg gctatgcgct cgtcaccatt tgggagcaca tgcgggaaac
     8581 ggggaaggtc aagttctcct acgagacgtt ccgctcgcac gccaggcggc acatcaaggc
     8641 caagcccgcc gatgtgcccg caccgcaggc caaggctgcg gaacccgcgc cggcacccaa
     8701 gacgccggag ccacggcggc cgaagcaggg gggcaaggct gaaaagccgg cccccgctgc
     8761 ggccccgacc ggcttcacct tcaacccaac accggacaaa aaggatcctc tacgccggac
     8821 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
     8881 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg
     8941 gtatggtggc ag
//
