LOCUS       V002576                 9619 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002576
VERSION     V002576
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9619)
  AUTHORS   Choi KH, Gaynor JB, White KG, Lopez C, Bosio CM, Karkhoff-Schweizer
            RR, Schweizer HP.
  TITLE     A Tn7-based broad-range bacterial cloning and expression system
  JOURNAL   Nat. Methods 2 (6), 443-448 (2005)
   PUBMED   15908923
REFERENCE   2  (bases 1 to 9619)
  AUTHORS   Choi K-H., Gaynor JB, White KG, Lopez CM, Karkhoff-Schweizer RK,
            Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-JAN-2005) Microbiology, Immunology and Pathology,
            Colorado State University, Fort Collins, CO 80523, USA
REFERENCE   3  (bases 1 to 9619)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9619)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat.
            Methods"; date: "2005"; volume: "2"; issue: "6"; pages: "443-448"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (12-JAN-2005) Microbiology, Immunology and Pathology, Colorado State
            University, Fort Collins, CO 80523, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(690..2216)
                     /codon_start=1
                     /gene="tnsD"
                     /product="Tn7 transposase subunit"
                     /label="tnsD"
                     /note="TnsD"
                     /protein_id="AAW78923.1"
                     /translation="MRNFPVPYSNELIYSTIARAGVYQGIVSPKQLLDEVYGNRKVVAT
                     LGLPSHLGVIARHLHQTGRYAVQQLIYEHTLFPLYAPFVGKERRDEAIRLMEYQAQGAV
                     HLMLGVAASRVKSDNRFRYCPDCVALQLNRYGEAFWQRDWYLPALPYCPKHGALVFFDR
                     AVDDHRHQFWALGHTELLSDYPKDSLSQLTALAAYIAPLLDAPRAQELSPSLEQWTLFY
                     QRLAQDLGLTKSKHIRHDLVAERVRQTFSDEALEKLDLKLAENKDTCWLKSIFRKHRKA
                     FSYLQHSIVWQALLPKLTVIEALQQASALTEHSITTRPVSQSVQPNSEDLSVKHKDWQQ
                     LVHKYQGIKAARQSLEGGVLYAWLYRHDRDWLVHWNQQHQQERLAPAPRVDWNQRDRIA
                     VRQLLRIIKRLDSSLDHPRATSSWLLKQTPNGTSLAKNLQKLPLVALCLKRYSESVEDY
                     QIRRISQAFIKLKQEDVELRRWRLLRSATLSKERITEEAQRFLEMVYGEE"
     gene            complement(690..2216)
                     /gene="tnsD"
                     /label="tnsD"
     CDS             complement(2222..3886)
                     /gene="tnsC"
                     /label="Transposon Tn7 transposition protein TnsC"
                     /note="Transposon Tn7 transposition protein TnsC from
                     Escherichia coli. Accession#: P05846"
     CDS             complement(3886..5991)
                     /gene="tnsB"
                     /label="Transposon Tn7 transposition protein TnsB"
                     /note="Transposon Tn7 transposition protein TnsB from
                     Escherichia coli. Accession#: P13989"
     mobile_element  6709..6933
                     /label="Tn7R"
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(7145..7161)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(7169..7185)
                     /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(7193..7223)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7238..7259)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     oriT            7635..7743
                     /label="oriT"
                     /note="incP origin of transfer"
     rep_origin      7766..8150
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(8562..9419)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9420..9524)
                     /label="AmpR promoter"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc ctgcaggtcg
      421 actctagagg atcccccgat tattttggct cgttgaagat ccgatgcttc aaactcaatc
      481 acacgatccg ccttatttat ttgctttgtt gtactgttaa ccaccttagc tcgacttaat
      541 aaaggcaaat gagtaaatcg agtccatttc cgttcttgca ttggattaaa ccaaacaaaa
      601 atacgccact ccttatgacc cgagttacgg aataaatgac caatatgtac aacctgcaca
      661 ttgtcattaa atgtagctag cctaaccact cactcttccc cataaaccat ttccaagaat
      721 ctttgtgctt cctcagttat ccgctcttta gataacgttg cacttcttaa taatcgccag
      781 cgcctaagct caacatcttc ctgtttaagc ttaataaaag cttggctaat ccgtctaatt
      841 tgataatctt ccacactctc tgagtaacgc tttaagcaaa gcgctaccaa aggcagtttc
      901 tgtagatttt ttgcaagaga ggttccgtta ggagtttgct ttaacagcca gctcgatgtc
      961 gctcttgggt gatcaaggct actatctaga cgctttatga ttcttaatag ttgtcgtaca
     1021 gcaattcgat ctctttggtt ccaatcaact ctaggggcgg gtgccagacg ctcttgttga
     1081 tgctgttgat tccagtgaac tagccaatcc ctgtcatgtc ggtaaagcca agcgtatagc
     1141 accccaccct ctaaagactg tcttgccgcc ttaattcctt ggtatttatg cactagttgc
     1201 tgccagtctt tatgcttaac agataaatct tcagagttag gttgcacaga ctggctaaca
     1261 ggtcttgtcg ttatagagtg ctcagtaaga gcacttgcct gctgtagcgc ttctataacc
     1321 gttagttttg gcaataaggc ttgccacaca atactatgct gtaaataact aaaggctttt
     1381 ctatgcttac ggaatatact tttcagccaa cacgtgtcct tgttctctgc caactttaaa
     1441 tccagtttct ctagtgcctc atcactaaaa gtctgcctca ctctctccgc caccaagtca
     1501 tgacgaatgt gcttgctttt ggttagccct agatcctgcg ctaagcgctg ataaaacagc
     1561 gtccactgct caaggcttgg ggaaagctct tgcgctcgtg gagcatctaa cagaggggct
     1621 atataagcag ctagtgctgt taattgagat agggagtctt tggggtagtc tgaaagcagc
     1681 tcagtatgac ccaaagccca aaattgatgt cggtgatcat ctacagctct atcaaagaag
     1741 actaaagcac cgtgttttgg acaatatggc aaagcgggca aataccaatc tcgttgccaa
     1801 aaggcttccc catacctatt tagctgaaga gcaacgcaat cagggcagta tctaaagcgg
     1861 ttatcgctct taactctaga agcagcgact cctagcatta aatgcaccgc accttgcgct
     1921 tggtactcca ttaaccgaat agcttcgtct cggcgctcct tgcctacaaa cggagcatat
     1981 aaagggaata aggtatgctc ataaataagc tgctgaacag cgtaacgtcc tgtttgatgt
     2041 agatgtcttg ctatcacacc taaatgcgag ggcagaccta aggtagcgac caccttgcgg
     2101 ttgccataca cctcatccaa cagctgctta ggactaacaa tcccttgata aacgcctgcc
     2161 cgtgcaatag tgctataaat cagctcattc gagtacggaa caggaaagtt tctcatggct
     2221 aacctgcttg tttaaataag ctcgccatat ccactattac ccctttccct tttaaccgtt
     2281 catgcatggt tttttcaggt tggcgttgtg aatagatata acgtaaatcc gtatcaggca
     2341 agctatccca gtcgctgggc ttgactacct ttatagccga aaccttttta ctcttggagg
     2401 gcttttctag ttctgacact accgtttcgc cttccatcaa ccactgcaaa acaagaggca
     2461 gtaacttttg tcttgtcatc gttggattct ggctaaacgc ttttttaata gtgggaatta
     2521 acaggcttga atcgtaatcc tctttcagca tcagatataa atgacgctga tcttcggtat
     2581 ctaactcttg aagggctttt tcttctggtg tttgttcttg tatcgctgcg atatctagct
     2641 gaagttggat taaccgttta tcaatctcgg gaacgactag atcagaataa cgagcaatgc
     2701 gttctgggat acccgagcgt aatgcctcta gcatggggtg cacaggcttt aactcatctt
     2761 gatacacttg ccgcaataaa ccagcggtaa tacgctcatt gcctaaagct agcgcacgga
     2821 gctgagcgag tacaaaaagt tttactacaa tgtccatcac tccttggctt agctcatacc
     2881 acacatcacg gacctcatcc gataacagcg catctttgcg ttgtaaaagc tgtaattgcc
     2941 aaagattatc cgtaaaagcg atccactctt gattgggctt tccacgttgc gtttgttgta
     3001 taggatccca gaatatagct ccaaaccctg cccctctacg tgcagaccgc aaatcagcct
     3061 caaaaatctc tcgtgcttta ggggtaccaa tcaacatcac tggtacgcca ataatattca
     3121 ccatcgtcac aaaaaagttc agcatctctt gagatccacc cgaacgagag cggcttaaat
     3181 gctgaatttc atcaataacc aacaacccta aagcatgtgc attggctatt tgcgacatca
     3241 aagccaacat ggtttctata ccatgacgtt ttaagccata acgacgctca tagttcgagc
     3301 ccaaggctcg atccaacgct ctgaaaaaat tcaagcagat ttcttttagc gaaccattat
     3361 gcgagcagtc tattttcaaa tacaccacct gctctacatt gagttcacga tggtaaatca
     3421 cctgaggata cgtggctaga atacgatgaa gagaggtcgt cttcccacta ccagaacaac
     3481 caattaacaa taagctttgt gccgtagatc gtgcctcctc aaagcgaaat gtctccaact
     3541 ctcccgtttg aacacgctca taaccatttt gtaaatgctt ttgtaaatct cctgttttag
     3601 gatttctgcc tacgtagcca cctcgaatca tgaccgaaat acgctcactt agtagcaaat
     3661 gcgtacctaa tggctgaaaa tagtcatctg gaatacgaca aatggtatga gctctgataa
     3721 cacgggactt ttgcaagtca gaggaagtaa gctgtaaaga ggatttcagt gatgcagcac
     3781 tattcactga ctcttgtaat ggtggtaagg cctcgataaa aggattatca cgataagcct
     3841 ctacccccgt atcacgataa actgcttgaa tccgggtagc actcatgact catccttttc
     3901 tggtggatcc tgaaataatt caggcacgta tgtgggtagg ctgtaatctt cttgatcatc
     3961 cgcttccact gcgttgaaag gaataacttt agcctcatca cctgaagagc ttggcttcaa
     4021 atgctccgca cgttttttac gctccgaggt cacggcttct tttttattag ttttaatctg
     4081 cttaatgcgt gttgatttgg gctcagtagt actgggcgtt aacttattcg ctttctgaat
     4141 ggtttgctga ataaacgcct caagctccct gcgtttagtt aactcatcct gcttcgcatt
     4201 ggctttattg tgtttttctt gtgcttgtat atcccaaacc tcccaaaatg agagaccttt
     4261 aaactgccga ctacgttccg tcagattaca gcgccaaaat acacggctgc caacttgcgg
     4321 aaacaaataa atcgtatcaa ccagcactgg gtcataagcc gcttctaaat gttgaggtct
     4381 agctatatca gtgctccgct gcaaccaacc ctcacgcaga atctctgacc ccgagtaata
     4441 caaaccccac aaattaacgc caaatgaaga aatagagacc tttcggcgag gcagtaacgc
     4501 tactcgcaac tgctcttgct ccacagccct taaactacct gtacgatgct gcatacccca
     4561 ttgccatagc tggacaggaa tagacggtaa atctgtagga aaatcagcat ctcgatcgta
     4621 tttatccatc accagatggt tatttctgaa taagatcgta cgcaaaataa tttgtgtgaa
     4681 ctcaaatacc gacagagatg catctaacct atagtctgtt tcaccatggc ttttgatccg
     4741 actgccctct acaatgccag gtgcaaagga cttaaactcg gcttgtagtg ttctaaaagt
     4801 gctttccact atgcctttag catcgccacg tctaggtgga gcactttcca ctcgcacatt
     4861 aaaactagaa actaaggctt cgacctgatg actcattaat tcgccacggt ccgctagcaa
     4921 cacatctggc aaacctacac acggccagtc tgagctacta atctcaatat catgctgggc
     4981 acaaatggcc gttttgtcag agcaagcatt tacaaaagcc tgcatcgcca ccacataaga
     5041 cggattttca aagccgatat aaaagcccgt gatcatccga ctaaacacat caatcacaat
     5101 gtaaagcgtt ggtcttccta tgattttttg gcgatcatga tgatccacta aataaatatc
     5161 agcaatcgtg gcatcaatct cataacgact cccagggcct aacgcctgag aagtggctgt
     5221 actacttaag ggtcgtacgt cttttttata tacccctgct ttaactctag actttaagcg
     5281 ctgagcttta gggtattctc gatcataaaa ataacgaaac tgacgtagtg ttgggtaatc
     5341 ctcttgggga atgcgaggaa aatactgagc aaacaagtcc acaaatcgtc tataggcaac
     5401 ggtggtcttt gtaccttttt gatttaacag gtgcttttct atggtcaacc taaaaaggcg
     5461 ttcaatctcg ggcgttacct tggttccttc acccttacca tattctctgg ctcggccaat
     5521 ctttgctgtt cctgtcgctg aacgtctttc ccctggtgca ccgctgtttt tgtagtcagg
     5581 aattaatgca ttaggcgttt gaccacgctg ccagtaacgg cgtaacaact tataaactgt
     5641 agccttagta accttatgtt cttggaccac atgctcaacg agttcgcttc tgactttagg
     5701 gtcgaaacga tccttactat taataatagg aagaattttt cgataatcct cctcgcgctt
     5761 ctggaagctg acagaatcta cagacggctc ttctaaatca agatgtacat aagggtcatc
     5821 agttctaact aagcggcctt catctaaata ctgcattagc aacaactcag ccctagcttg
     5881 tggaactcct ttatcagcgc ttatttgcat ccagacaact tggccatcct ctatagccaa
     5941 aatgcgatac ggatcattat caaatagcac aacctcatta atttgccaca tagcgcaact
     6001 cctccatatt cactacttgg ctaatacaga gatctgcaca cttatttgcc ctgaaagact
     6061 tataaatatt gaacttaata aaaccatttg cggttaaggc tcgtatctca ctcaatgttt
     6121 tgcctaactc caaatcataa gcaatatcaa cctgcttaca gacattgata atgttttcat
     6181 ctcctttttc ttgcaggata tgggccaatg gggatagttg tgctaaaagc tccgcagaaa
     6241 cttcttctgt tttcactgaa taaagccatt caatattttc ttttactacg ggatttattt
     6301 ctttatcagt aaaaatgaac caaggaattt gcttttgctg ccaatagcga cgctctagtt
     6361 ctagtttttc taaggtacgc tcgtcttgta aggctgctgc aggtttgact tgaatagcaa
     6421 actgctcaaa aggaccatct ttgcagtcca ctaaaaaatc agtagacata acctgatcta
     6481 caccacgaat aacaggatgc ttaataccac tatctattgc aatctgcctg gtatcactag
     6541 gtaataaggg gaactgctcg cgtatatcta gcacgctgct ctcccactca agactgagaa
     6601 aaacagcaag ctctaagtca gatagcaaat gatggactcg tcccgtctta tgagaataaa
     6661 tacggtggga acgacctgaa gaaggaactt cttgtactgt tagccatgga atatagtctt
     6721 taccatgccc ttggccacgc ccctctttaa tacgacgggc aatttgcact tcagaaaatg
     6781 aagagtttgc tttagccata acaaaagtcc agtatgcttt ttcacagcat aactggactg
     6841 atttcagttt acaactattc tgtctagttt aagactttat tgtcatagtt tagatctatt
     6901 ttgttccctt taagacttta ttgtccgccc acacccgctt acgcagggca tccatttatt
     6961 actcaaccgt aaccgatttt gccaggttac gcggctggtc aacgtcggtg cctttgatca
     7021 gcgcgacatg gtaagccagc agctgcagcg gaacggtgta gaagatcggt gcaatcacct
     7081 cttccacatg cggcatctcg actctagagg atccccgggt accgagctcg aattcgtaat
     7141 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     7201 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     7261 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     7321 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     7381 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     7441 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg ttaaggtata
     7501 ctttccgctg cataaccctg cttcggggtc attatagcga ttttttcggt atatccatcc
     7561 tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc ttggtgtatc
     7621 caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc
     7681 ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct gcgaggctgg
     7741 ccgataagct ctgataacgc aggaaagaac atgggcagtt caacctgttg atagtacgta
     7801 ctaagctctc atgtttcacg tactaagctc tcaagtttaa cgtactaagc tctcatgttt
     7861 aacgaactaa accctcatgg ctaacgtact aagctctcat ggctaacgta ctaagctctc
     7921 atgtttcacg tactaagctc tcatgtttga acaataaaat taatataaat cagcaactta
     7981 aatagcctct aaggttttaa gttttataag aaaaaaaaga atatataagg cttttaaagc
     8041 ttttaaggtt taacggttgt ggacaacaag ccagggatgt aacgcactga gaagccctta
     8101 gagcctctca aagcaatttt gagtgacaca ggaacactta acggctgaca accggctggt
     8161 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     8221 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     8281 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     8341 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     8401 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     8461 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     8521 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     8581 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8641 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     8701 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8761 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8821 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8881 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8941 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     9001 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     9061 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     9121 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     9181 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     9241 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     9301 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     9361 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     9421 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     9481 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     9541 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     9601 atcacgaggc cctttcgtc
//
