LOCUS       40924_20835        7657 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Transposon mutagenesis vector pG18-STM, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7657)
  AUTHORS   Pobigaylo N, Wetter D, Szymczak S, Schiller U, Kurtz S, Meyer F, 
            Nattkemper TW, Becker A.
  TITLE     Construction of a large signature-tagged mini-Tn5 transposon library
            and its application to mutagenesis of Sinorhizobium meliloti
  JOURNAL   Appl. Environ. Microbiol. 72 (6), 4329-4337 (2006)
  PUBMED    16751548
REFERENCE   2  (bases 1 to 7657)
  AUTHORS   Pobigaylo N, Becker A.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-FEB-2006) Department of Genetics, University of 
            Bielefeld, Universitaetsstrasse 25, Bielefeld D-33615, Germany
REFERENCE   3  (bases 1 to 7657)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7657)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2006"; volume: "72"; issue: "6"; pages:
            "4329-4337"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (20-FEB-2006) Department of Genetics, University of Bielefeld, 
            Universitaetsstrasse 25, Bielefeld D-33615, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    145..468
                     /label=similar to transposase
                     /note="similar to transposase"
     CDS             complement(509..1039)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(1228..1256)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     oriT            complement(1479..1587)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      1799..2387
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2675..2696
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2711..2741
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2749..2765
                     /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)."
     repeat_region   2777..2795
                     /note="Tn5 outside end; required for transposition"
     CDS             2876..4684
                     /label=GUS
                     /note="beta-glucuronidase"
     promoter        4759..4787
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    4795..4811
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     CDS             4863..5654
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    5791..6033
                     /note="artificial linker for cloning and amplification of 
                     signature tags"
     gap             5855..5878
                     /estimated_length=24
     misc_feature    5855..5878
                     /note="h-tag; variable region, signature tag"
     gap             5946..5969
                     /estimated_length=24
     misc_feature    5946..5969
                     /note="k-tag; variable region, signature tag"
     repeat_region   6034..6052
                     /note="Tn5 inside end; required for transposition"
     CDS             6139..7566
                     /label=Tn5 transposase
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"
     primer_bind     complement(7636..7652)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
       61 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
      121 ctgaatggcg aatggcgata agctagcttc acgctgccgc aagcactcag ggcgcaaggg
      181 ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag aaacggtgct gaccccggat
      241 gaatgtcagc tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt
      301 agcttgcagt gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga
      361 accggaattg ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg
      421 gatggctttc ttgccgccaa ggatctgatg gcgcagggga tcaagatccg tcggccggga
      481 agccgatctc ggcttgaacg aattgttagg tggcggtact tgggtcgata tcaaagtgca
      541 tcacttcttc ccgtatgccc aactttgtat agagagccac tgcgggatcg tcaccgtaat
      601 ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg aggagattga
      661 tgagcgcggt ggcaatgccc tgcctccggt gctcgccgga gactgcgaga tcatagatat
      721 agatctcact acgcggctgc tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa
      781 ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt actacggagc aagttcccga
      841 ggtaatcgga gtccggctga tgttgggagt aggtggctac gtctccgaac tcacgaccga
      901 aaagatcaag agcagcccgc atggatttga cttggtcagg gccgagccta catgtgcgaa
      961 tgatgcccat acttgagcca cctaactttg ttttagggcg actgccctgc tgcgtaacat
     1021 cgttgctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa
     1081 cgcgcttgct gcttggatgc ccgaggcata gactgtacaa aaaaacagtc ataacaagcc
     1141 atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg
     1201 cgtgagcgca tacgctactt gcattacagt ttacgaaccg aacaggctta tgtcaactgg
     1261 gttcgtgcct tcatccgttt ccacggtgtg cgtcacccgg caaccttggg cagcagcgaa
     1321 gtcgaggcat ttctgtcctg gctggcgaac gagcgcaagg tttcggtctc cacgcatcgt
     1381 caggcattgg cggccttgct gttcttctac ggcaaggtgc tgtgcacgga tctgccctgg
     1441 cttcaggaga tcggaagacc tcggccgacg gatctgatcg gccagcctcg cagagcagga
     1501 ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg
     1561 ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct
     1621 acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa
     1681 atcgctataa tgaccccgaa gcagggttat gcagcggaaa agccatgacc aaaatccctt
     1741 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1801 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1861 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1921 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     1981 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2041 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2101 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2161 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2221 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2281 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2341 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2401 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2461 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2521 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     2581 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     2641 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     2701 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     2761 aacaatttgt ggaattctga ctcttataca caagtgcggc cgcgcgaatt gatccggtga
     2821 ttgattgagc aagctaattc cggaccagta ttattatctt aatgaggagt cccttatgtt
     2881 acgtcctgta gaaaccccaa cccgtgaaat caaaaaactc gacggcctgt gggcattcag
     2941 tctggatcgc gaaaactgtg gaattgatca gcgttggtgg gaaagcgcgt tacaagaaag
     3001 ccgggcaatt gctgtgccag gcagttttaa cgatcagttc gccgatgcag atattcgtaa
     3061 ttatgcgggc aacgtctggt atcagcgcga agtctttata ccgaaaggtt gggcaggcca
     3121 gcgtatcgtg ctgcgtttcg atgcggtcac tcattacggc aaagtgtggg tcaataatca
     3181 ggaagtgatg gagcatcagg gcggctatac gccatttgaa gccgatgtca cgccgtatgt
     3241 tattgccggg aaaagtgtac gtatcaccgt ttgtgtgaac aacgaactga actggcagac
     3301 tatcccgccg ggaatggtga ttaccgacga aaacggcaag aaaaagcagt cttacttcca
     3361 tgatttcttt aactatgccg ggatccatcg cagcgtaatg ctctacacca cgccgaacac
     3421 ctgggtggac gatatcaccg tggtgacgca tgtcgcgcaa gactgtaacc acgcgtctgt
     3481 tgactggcag gtggtggcca atggtgatgt cagcgttgaa ctgcgtgatg cggatcaaca
     3541 ggtggttgca actggacaag gcactagcgg gactttgcaa gtggtgaatc cgcacctctg
     3601 gcaaccgggt gaaggttatc tctatgaact gtgcgtcaca gccaaaagcc agacagagtg
     3661 tgatatctac ccgcttcgcg tcggcatccg gtcagtggca gtgaagggcg aacagttcct
     3721 gattaaccac aaaccgttct actttactgg ctttggtcgt catgaagatg cggacttacg
     3781 tggcaaagga ttcgataacg tgctgatggt gcacgaccac gcattaatgg actggattgg
     3841 ggccaactcc taccgtacct cgcattaccc ttacgctgaa gagatgctcg actgggcaga
     3901 tgaacatggc atcgtggtga ttgatgaaac tgctgctgtc ggctttaacc tctctttagg
     3961 cattggtttc gaagcgggca acaagccgaa agaactgtac agcgaagagg cagtcaacgg
     4021 ggaaactcag caagcgcact tacaggcgat taaagagctg atagcgcgtg acaaaaacca
     4081 cccaagcgtg gtgatgtgga gtattgccaa cgaaccggat acccgtccgc aaggtgcacg
     4141 ggaatatttc gcgccactgg cggaagcaac gcgtaaactc gacccgacgc gtccgatcac
     4201 ctgcgtcaat gtaatgttct gcgacgctca caccgatacc atcagcgatc tctttgatgt
     4261 gctgtgcctg aaccgttatt acggatggta tgtccaaagc ggcgatttgg aaacggcaga
     4321 gaaggtactg gaaaaagaac ttctggcctg gcaggagaaa ctgcatcagc cgattatcat
     4381 caccgaatac ggcgtggata cgttagccgg gctgcactca atgtacaccg acatgtggag
     4441 tgaagagtat cagtgtgcat ggctggatat gtatcaccgc gtctttgatc gcgtcagcgc
     4501 cgtcgtcggt gaacaggtat ggaatttcgc cgattttgcg acctcgcaag gcatattgcg
     4561 cgttggcggt aacaagaaag ggatcttcac tcgcgaccgc aaaccgaagt cggcggcttt
     4621 tctgctgcaa aaacgctgga ctggcatgaa cttcggtgaa aaaccgcagc agggaggcaa
     4681 acaatgaatc aacaactctc ctggcgcacc atcgtcggct acagcctcgg tgaagctagc
     4741 ttactcccca tccccctgtt gacaattaat catcggctcg tataatgtgt ggaattgtga
     4801 gcggataaca atttcacaca ggaaacagga tcgatcaaga gacaggatga ggatcgtttc
     4861 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4921 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4981 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5041 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5101 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5161 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5221 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5281 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5341 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5401 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5461 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5521 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5581 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5641 ttgacgagtt cttctgagcg ggactctggg gaattgatcc ggtgattgat tgagcaagct
     5701 gatccgtcga tcgacctgca tctagcccgc ctaatgagcg ggcttttttt tagatgcagc
     5761 caagctagct tgcggccagc ttgcggccgc ggcctaggcg gcctagtcct ggtgcattga
     5821 gccctactag ctctacgacg gtccacctaa gcttnnnnnn nnnnnnnnnn nnnnnnnnaa
     5881 gcttgcggcg gaggggagtt aaaatgaaat cacgcagcta tatgaatgcc gccacccccg
     5941 gtaccnnnnn nnnnnnnnnn nnnnnnnnng gtaccttacc gtacttcctt aaactacgtg
     6001 ccccgtaaac cacgtccttt ggcctaggcg gccagatctg atcaagagac agtctagaca
     6061 caagtagcgt cctgaacgga acctttcccg ttttccagga tctgatcttc catgtgagct
     6121 cctaacatgg taacgttcat gataacttct gctcttcatc gtgcggccga ctgggctaaa
     6181 tctgtgttct cttcggcggc gctgggtgat cctcgccgta ctgcccgctt ggttaacgtc
     6241 gccgcccaat tggcaaaata ttctggtaaa tcaataacca tctcatcaga gggtagtgaa
     6301 gccatgcagg aaggcgctta ccgatttatc cgcaatccca acgtttctgc cgaggcgatc
     6361 agaaaggctg gcgccatgca aacagtcaag ttggctcagg agtttcccga actgctggcc
     6421 attgaggaca ccacctcttt gagttatcgc caccaggtcg ccgaagagct tggcaagctg
     6481 ggctctattc aggataaatc ccgcggatgg tgggttcact ccgttctctt gctcgaggcc
     6541 accacattcc gcaccgtagg attactgcat caggagtggt ggatgcgccc ggatgaccct
     6601 gccgatgcgg atgaaaagga gagtggcaaa tggctggcag ccgccgcaac tagccggtta
     6661 cgcatgggca gcatgatgag caacgtgatt gcggtctgtg accgcgaagc cgatattcat
     6721 gcttatctgc aggacaaact ggcgcataac gagcgcttcg tggtgcgctc caagcaccca
     6781 cgcaaggacg tagagtctgg gttgtatctg tacgaccatc tgaagaacca accggagttg
     6841 ggtggctatc agatcagcat tccgcaaaag ggcgtggtgg ataaacgcgg taaacgtaaa
     6901 aatcgaccag cccgcaaggc gagcttgagc ctgcgcagtg ggcgcatcac gctaaaacag
     6961 gggaatatca cgctcaacgc ggtgctggcc gaggagatta acccgcccaa gggtgagacc
     7021 ccgttgaaat ggttgttgct gaccagcgaa ccggtcgagt cgctagccca agccttgcgc
     7081 gtcatcgaca tttataccca tcgctggcgg atcgaggagt tccataaggc atggaaaacc
     7141 ggagcaggag ccgagaggca acgcatggag gagccggata atctggagcg gatggtctcg
     7201 atcctctcgt ttgttgcggt caggctgtta cagctcagag agagcttcac gctgccgcaa
     7261 gcactcaggg cgcaagggct gctaaaggaa gcggaacacg tagaaagcca gtccgcagaa
     7321 acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag
     7381 cgcaaagaga aagcaggtag cttgcagtgg gcctacatgg cgatagctag actgggcggt
     7441 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggga aggttgggaa
     7501 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
     7561 aagatctgat cgcatgcgat cagatcttga tccctataac gtgaagctct ctctgagcat
     7621 gcaagctagc ttggcactgg ccgtcgtttt acaacgt
//
