LOCUS       Exported                9983 bp DNA     circular SYN 19-JUN-2024
DEFINITION  Exported.
ACCESSION   V002574
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9983)
  AUTHORS   Choi KH, Mima T, Casart Y, Rholl D, Kumar A, Beacham IR, Schweizer 
            HP.
  TITLE     Genetic tools for select-agent-compliant manipulation of 
            Burkholderia pseudomallei
  JOURNAL   Appl. Environ. Microbiol. 74 (4), 1064-1075 (2008)
  PUBMED    18156318
REFERENCE   2  (bases 1 to 9983)
  AUTHORS   Choi K-H., Rholl D, Schweizer HP.
  TITLE     A Tn7-based broad-host bacterial cloning and expression system
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 9983)
  AUTHORS   Choi K-H., Rholl D, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-SEP-2007) Microbiology, Immunology and Pathology, 
            Colorado State University, 1682 Campus Delivery, Ft. Collins, CO 
            80523-1682, USA
REFERENCE   4  (bases 1 to 9983)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 9983)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 9983)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2008"; volume: "74"; issue: "4"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (21-SEP-2007) Microbiology, Immunology and Pathology, Colorado State
            University, 1682 Campus Delivery, Ft. Collins, CO 80523-1682, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     SGRef: number: 5; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      complement(1470..1854)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            complement(1877..1985)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     protein_bind    2361..2382
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2397..2427
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2435..2451
                     /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)."
     primer_bind     2459..2475
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2512..2540
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     mobile_element  complement(3051..3275)
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             3993..6098
                     /gene="tnsB"
                     /label=Transposon Tn7 transposition protein TnsB
                     /note="Transposon Tn7 transposition protein TnsB from 
                     Escherichia coli. Accession#: P13989"
     CDS             6098..7762
                     /gene="tnsC"
                     /label=Transposon Tn7 transposition protein TnsC
                     /note="Transposon Tn7 transposition protein TnsC from 
                     Escherichia coli. Accession#: P05846"
     CDS             7768..9294
                     /codon_start=1
                     /gene="tnsD"
                     /product="TnsD"
                     /label=tnsD
                     /note="Tn7 transposase subunit"
                     /protein_id="ABW35346.1"
                     /translation="MRNFPVPYSNELIYSTIARAGVYQGIVSPKQLLDEVYGNRKVVAT
                     LGLPSHLGVIARHLHQTGRYAVQQLIYEHTLFPLYAPFVGKERRDEAIRLMEYQAQGAV
                     HLMLGVAASRVKSDNRFRYCPDCVALQLNRYGEAFWQRDWYLPALPYCPKHGALVFFDR
                     AVDDHRHQFWALGHTELLSDYPKDSLSQLTALAAYIAPLLDAPRAQELSPSLEQWTLFY
                     QRLAQDLGLTKSKHIRHDLVAERVRQTFSDEALEKLDLKLAENKDTCWLKSIFRKHRKA
                     FSYLQHSIVWQALLPKLTVIEALQQASALTEHSITTRPVSQSVQPNSEDLSVKHKDWQQ
                     LVHKYQGIKAARQSLEGGVLYAWLYRHDRDWLVHWNQQHQQERLAPAPRVDWNQRDRIA
                     VRQLLRIIKRLDSSLDHPRATSSWLLKQTPNGTSLAKNLQKLPLVALCLKRYSESVEDY
                     QIRRISQAFIKLKQEDVELRRWRLLRSATLSKERITEEAQRFLEMVYGEE"
     gene            7768..9294
                     /gene="tnsD"
                     /label=tnsD
     primer_bind     complement(9589..9605)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagccggtt gtcagccgtt aagtgttcct gtgtcactca
     1501 aaattgcttt gagaggctct aagggcttct cagtgcgtta catccctggc ttgttgtcca
     1561 caaccgttaa accttaaaag ctttaaaagc cttatatatt cttttttttc ttataaaact
     1621 taaaacctta gaggctattt aagttgctga tttatattaa ttttattgtt caaacatgag
     1681 agcttagtac gtgaaacatg agagcttagt acgttagcca tgagagctta gtacgttagc
     1741 catgagggtt tagttcgtta aacatgagag cttagtacgt taaacttgag agcttagtac
     1801 gtgaaacatg agagcttagt acgtactatc aacaggttga actgcccatg ttctttcctg
     1861 cgttatcaga gcttatcggc cagcctcgca gagcaggatt cccgttgagc accgccaggt
     1921 gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc
     1981 tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc
     2041 ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc
     2101 agggttatgc agcggaaagt ataccttaac atgttctttc ctgcgttatc ccctgattct
     2161 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     2221 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     2281 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     2341 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     2401 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     2461 ggaaacagct atgaccatga ttacgaattc ggcttgaatt cacgaaccca gttgacataa
     2521 gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga
     2581 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
     2641 tgtttttttg tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
     2701 cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca
     2761 aagttaggca gccgttgtgc tggtgctttc tgatagttgt tgtggggtag gcagtcagag
     2821 ctcgatttgc ttgtcgccat aatagattca caagaaggat tcgacccggg gatcctctag
     2881 agtcgagatg ccgcatgtgg aagaggtgat tgcaccgatc ttctacaccg ttccgctgca
     2941 gctgctggct taccatgtcg cgctgatcaa aggcaccgac gttgaccagc cgcgtaacct
     3001 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     3061 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     3121 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     3181 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     3241 ggggcgtggc caagggcatg gtaaagacta tattccatgg ctaacagtac aagaagttcc
     3301 ttcttcaggt cgttcccacc gtatttattc tcataagacg ggacgagtcc atcatttgct
     3361 atctgactta gagcttgctg tttttctcag tcttgagtgg gagagcagcg tgctagatat
     3421 acgcgagcag ttccccttat tacctagtga taccaggcag attgcaatag atagtggtat
     3481 taagcatcct gttattcgtg gtgtagatca ggttatgtct actgattttt tagtggactg
     3541 caaagatggt ccttttgagc agtttgctat tcaagtcaaa cctgcagcag ccttacaaga
     3601 cgagcgtacc ttagaaaaac tagaactaga gcgtcgctat tggcagcaaa agcaaattcc
     3661 ttggttcatt tttactgata aagaaataaa tcccgtagta aaagaaaata ttgaatggct
     3721 ttattcagtg aaaacagaag aagtttctgc ggagctttta gcacaactat ccccattggc
     3781 ccatatcctg caagaaaaag gagatgaaaa cattatcaat gtctgtaagc aggttgatat
     3841 tgcttatgat ttggagttag gcaaaacatt gagtgagata cgagccttaa ccgcaaatgg
     3901 ttttattaag ttcaatattt ataagtcttt cagggcaaat aagtgtgcag atctctgtat
     3961 tagccaagta gtgaatatgg aggagttgcg ctatgtggca aattaatgag gttgtgctat
     4021 ttgataatga tccgtatcgc attttggcta tagaggatgg ccaagttgtc tggatgcaaa
     4081 taagcgctga taaaggagtt ccacaagcta gggctgagtt gttgctaatg cagtatttag
     4141 atgaaggccg cttagttaga actgatgacc cttatgtaca tcttgattta gaagagccgt
     4201 ctgtagattc tgtcagcttc cagaagcgcg aggaggatta tcgaaaaatt cttcctatta
     4261 ttaatagtaa ggatcgtttc gaccctaaag tcagaagcga actcgttgag catgtggtcc
     4321 aagaacataa ggttactaag gctacagttt ataagttgtt acgccgttac tggcagcgtg
     4381 gtcaaacgcc taatgcatta attcctgact acaaaaacag cggtgcacca ggggaaagac
     4441 gttcagcgac aggaacagca aagattggcc gagccagaga atatggtaag ggtgaaggaa
     4501 ccaaggtaac gcccgagatt gaacgccttt ttaggttgac catagaaaag cacctgttaa
     4561 atcaaaaagg tacaaagacc accgttgcct atagacgatt tgtggacttg tttgctcagt
     4621 attttcctcg cattccccaa gaggattacc caacactacg tcagtttcgt tatttttatg
     4681 atcgagaata ccctaaagct cagcgcttaa agtctagagt taaagcaggg gtatataaaa
     4741 aagacgtacg acccttaagt agtacagcca cttctcaggc gttaggccct gggagtcgtt
     4801 atgagattga tgccacgatt gctgatattt atttagtgga tcatcatgat cgccaaaaaa
     4861 tcataggaag accaacgctt tacattgtga ttgatgtgtt tagtcggatg atcacgggct
     4921 tttatatcgg ctttgaaaat ccgtcttatg tggtggcgat gcaggctttt gtaaatgctt
     4981 gctctgacaa aacggccatt tgtgcccagc atgatattga gattagtagc tcagactggc
     5041 cgtgtgtagg tttgccagat gtgttgctag cggaccgtgg cgaattaatg agtcatcagg
     5101 tcgaagcctt agtttctagt tttaatgtgc gagtggaaag tgctccacct agacgtggcg
     5161 atgctaaagg catagtggaa agcactttta gaacactaca agccgagttt aagtcctttg
     5221 cacctggcat tgtagagggc agtcggatca aaagccatgg tgaaacagac tataggttag
     5281 atgcatctct gtcggtattt gagttcacac aaattatttt gcgtacgatc ttattcagaa
     5341 ataaccatct ggtgatggat aaatacgatc gagatgctga ttttcctaca gatttaccgt
     5401 ctattcctgt ccagctatgg caatggggta tgcagcatcg tacaggtagt ttaagggctg
     5461 tggagcaaga gcagttgcga gtagcgttac tgcctcgccg aaaggtctct atttcttcat
     5521 ttggcgttaa tttgtggggt ttgtattact cggggtcaga gattctgcgt gagggttggt
     5581 tgcagcggag cactgatata gctagacctc aacatttaga agcggcttat gacccagtgc
     5641 tggttgatac gatttatttg tttccgcaag ttggcagccg tgtattttgg cgctgtaatc
     5701 tgacggaacg tagtcggcag tttaaaggtc tctcattttg ggaggtttgg gatatacaag
     5761 cacaagaaaa acacaataaa gccaatgcga agcaggatga gttaactaaa cgcagggagc
     5821 ttgaggcgtt tattcagcaa accattcaga aagcgaataa gttaacgccc agtactactg
     5881 agcccaaatc aacacgcatt aagcagatta aaactaataa aaaagaagcc gtgacctcgg
     5941 agcgtaaaaa acgtgcggag catttgaagc caagctcttc aggtgatgag gctaaagtta
     6001 ttcctttcaa cgcagtggaa gcggatgatc aagaagatta cagcctaccc acatacgtgc
     6061 ctgaattatt tcaggatcca ccagaaaagg atgagtcatg agtgctaccc ggattcaagc
     6121 agtttatcgt gatacggggg tagaggctta tcgtgataat ccttttatcg aggccttacc
     6181 accattacaa gagtcagtga atagtgctgc atcactgaaa tcctctttac agcttacttc
     6241 ctctgacttg caaaagtccc gtgttatcag agctcatacc atttgtcgta ttccagatga
     6301 ctattttcag ccattaggta cgcatttgct actaagtgag cgtatttcgg tcatgattcg
     6361 aggtggctac gtaggcagaa atcctaaaac aggagattta caaaagcatt tacaaaatgg
     6421 ttatgagcgt gttcaaacgg gagagttgga gacatttcgc tttgaggagg cacgatctac
     6481 ggcacaaagc ttattgttaa ttggttgttc tggtagtggg aagacgacct ctcttcatcg
     6541 tattctagcc acgtatcctc aggtgattta ccatcgtgaa ctcaatgtag agcaggtggt
     6601 gtatttgaaa atagactgct cgcataatgg ttcgctaaaa gaaatctgct tgaatttttt
     6661 cagagcgttg gatcgagcct tgggctcgaa ctatgagcgt cgttatggct taaaacgtca
     6721 tggtatagaa accatgttgg ctttgatgtc gcaaatagcc aatgcacatg ctttagggtt
     6781 gttggttatt gatgaaattc agcatttaag ccgctctcgt tcgggtggat ctcaagagat
     6841 gctgaacttt tttgtgacga tggtgaatat tattggcgta ccagtgatgt tgattggtac
     6901 ccctaaagca cgagagattt ttgaggctga tttgcggtct gcacgtagag gggcagggtt
     6961 tggagctata ttctgggatc ctatacaaca aacgcaacgt ggaaagccca atcaagagtg
     7021 gatcgctttt acggataatc tttggcaatt acagctttta caacgcaaag atgcgctgtt
     7081 atcggatgag gtccgtgatg tgtggtatga gctaagccaa ggagtgatgg acattgtagt
     7141 aaaacttttt gtactcgctc agctccgtgc gctagcttta ggcaatgagc gtattaccgc
     7201 tggtttattg cggcaagtgt atcaagatga gttaaagcct gtgcacccca tgctagaggc
     7261 attacgctcg ggtatcccag aacgcattgc tcgttattct gatctagtcg ttcccgagat
     7321 tgataaacgg ttaatccaac ttcagctaga tatcgcagcg atacaagaac aaacaccaga
     7381 agaaaaagcc cttcaagagt tagataccga agatcagcgt catttatatc tgatgctgaa
     7441 agaggattac gattcaagcc tgttaattcc cactattaaa aaagcgttta gccagaatcc
     7501 aacgatgaca agacaaaagt tactgcctct tgttttgcag tggttgatgg aaggcgaaac
     7561 ggtagtgtca gaactagaaa agccctccaa gagtaaaaag gtttcggcta taaaggtagt
     7621 caagcccagc gactgggata gcttgcctga tacggattta cgttatatct attcacaacg
     7681 ccaacctgaa aaaaccatgc atgaacggtt aaaagggaaa ggggtaatag tggatatggc
     7741 gagcttattt aaacaagcag gttagccatg agaaactttc ctgttccgta ctcgaatgag
     7801 ctgatttata gcactattgc acgggcaggc gtttatcaag ggattgttag tcctaagcag
     7861 ctgttggatg aggtgtatgg caaccgcaag gtggtcgcta ccttaggtct gccctcgcat
     7921 ttaggtgtga tagcaagaca tctacatcaa acaggacgtt acgctgttca gcagcttatt
     7981 tatgagcata ccttattccc tttatatgct ccgtttgtag gcaaggagcg ccgagacgaa
     8041 gctattcggt taatggagta ccaagcgcaa ggtgcggtgc atttaatgct aggagtcgct
     8101 gcttctagag ttaagagcga taaccgcttt agatactgcc ctgattgcgt tgctcttcag
     8161 ctaaataggt atggggaagc cttttggcaa cgagattggt atttgcccgc tttgccatat
     8221 tgtccaaaac acggtgcttt agtcttcttt gatagagctg tagatgatca ccgacatcaa
     8281 ttttgggctt tgggtcatac tgagctgctt tcagactacc ccaaagactc cctatctcaa
     8341 ttaacagcac tagctgctta tatagcccct ctgttagatg ctccacgagc gcaagagctt
     8401 tccccaagcc ttgagcagtg gacgctgttt tatcagcgct tagcgcagga tctagggcta
     8461 accaaaagca agcacattcg tcatgacttg gtggcggaga gagtgaggca gacttttagt
     8521 gatgaggcac tagagaaact ggatttaaag ttggcagaga acaaggacac gtgttggctg
     8581 aaaagtatat tccgtaagca tagaaaagcc tttagttatt tacagcatag tattgtgtgg
     8641 caagccttat tgccaaaact aacggttata gaagcgctac agcaggcaag tgctcttact
     8701 gagcactcta taacgacaag acctgttagc cagtctgtgc aacctaactc tgaagattta
     8761 tctgttaagc ataaagactg gcagcaacta gtgcataaat accaaggaat taaggcggca
     8821 agacagtctt tagagggtgg ggtgctatac gcttggcttt accgacatga cagggattgg
     8881 ctagttcact ggaatcaaca gcatcaacaa gagcgtctgg cacccgcccc tagagttgat
     8941 tggaaccaaa gagatcgaat tgctgtacga caactattaa gaatcataaa gcgtctagat
     9001 agtagccttg atcacccaag agcgacatcg agctggctgt taaagcaaac tcctaacgga
     9061 acctctcttg caaaaaatct acagaaactg cctttggtag cgctttgctt aaagcgttac
     9121 tcagagagtg tggaagatta tcaaattaga cggattagcc aagcttttat taagcttaaa
     9181 caggaagatg ttgagcttag gcgctggcga ttattaagaa gtgcaacgtt atctaaagag
     9241 cggataactg aggaagcaca aagattcttg gaaatggttt atggggaaga gtgagtggtt
     9301 aggctagcta catttaatga caatgtgcag gttgtacata ttggtcattt attccgtaac
     9361 tcgggtcata aggagtggcg tatttttgtt tggtttaatc caatgcaaga acggaaatgg
     9421 actcgattta ctcatttgcc tttattaagt cgagctaagg tggttaacag tacaacaaag
     9481 caaataaata aggcggatcg tgtgattgag tttgaagcat cggatcttca acgagccaaa
     9541 ataatcgggg gatcctctag agtcgacctg caggcatgca agcttggcac tggccgtcgt
     9601 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     9661 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     9721 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     9781 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     9841 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     9901 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     9961 accgtcatca ccgaaacgcg cga
//
