LOCUS       sequence-435439-        9615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn7 transposase expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9615)
  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. 2005 Jun;2(6):443-8.
  PUBMED    15908923
REFERENCE   2  (bases 1 to 9615)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods."; date: "2005-06"; volume: "2(6)"; pages: "443-8"
COMMENT     Sequence Label: pTNS2
FEATURES             Location/Qualifiers
     source          1..9615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     29..47
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(85..107)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     207..226
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     420..442
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     434..451
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     435..451
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     mobile_element  6763..6987
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(7198..7214)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7198..7214)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7211..7233)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7222..7238
                     /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)."
     promoter        complement(join(7246..7252,7253..7270,7271..7276))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7291..7312
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7429..7446)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     oriT            7687..7795
                     /note="incP origin of transfer"
     rep_origin      7846..8202
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(join(8611..9402,9403..9471))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9234..9253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9472..9576)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
       61 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
      121 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      181 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
      241 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
      301 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      361 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      421 ccagtcacga cgttgtaaaa cgacggccag tgccaagctt gcatgcctgc aggtcgactc
      481 tagaggatcc cgattatttt ggctcgttga agatccgatg cttcaaactc aatcacacga
      541 tccgccttat ttatttgctt tgttgtactg ttaaccacct tagctcgact taataaaggc
      601 aaatgagtaa atcgagtcca tttccgttct tgcattggat taaaccaaac aaaaatacgc
      661 cactccttat gacccgagtt acggaataaa tgaccaatat gtacaacctg cacattgtca
      721 ttaaatgtag ctagcctaac cactcactct tccccataaa ccatttccaa gaatctttgt
      781 gcttcctcag ttatccgctc tttagataac gttgcacttc ttaataatcg ccagcgccta
      841 agctcaacat cttcctgttt aagcttaata aaagcttggc taatccgtct aatttgataa
      901 tcttccacac tctctgagta acgctttaag caaagcgcta ccaaaggcag tttctgtaga
      961 ttttttgcaa gagaggttcc gttaggagtt tgctttaaca gccagctcga tgtcgctctt
     1021 gggtgatcaa ggctactatc tagacgcttt atgattctta atagttgtcg tacagcaatt
     1081 cgatctcttt ggttccaatc aactctaggg gcgggtgcca gacgctcttg ttgatgctgt
     1141 tgattccagt gaactagcca atccctgtca tgtcggtaaa gccaagcgta tagcacccca
     1201 ccctctaaag actgtcttgc cgccttaatt ccttggtatt tatgcactag ttgctgccag
     1261 tctttatgct taacagataa atcttcagag ttaggttgca cagactggct aacaggtctt
     1321 gtcgttatag agtgctcagt aagagcactt gcctgctgta gcgcttctat aaccgttagt
     1381 tttggcaata aggcttgcca cacaatacta tgctgtaaat aactaaaggc ttttctatgc
     1441 ttacggaata tacttttcag ccaacacgtg tccttgttct ctgccaactt taaatccagt
     1501 ttctctagtg cctcatcact aaaagtctgc ctcactctct ccgccaccaa gtcatgacga
     1561 atgtgcttgc ttttggttag ccctagatcc tgcgctaagc gctgataaaa cagcgtccac
     1621 tgctcaaggc ttggggaaag ctcttgcgct cgtggagcat ctaacagagg ggctatataa
     1681 gcagctagtg ctgttaattg agatagggag tctttggggt agtctgaaag cagctcagta
     1741 tgacccaaag cccaaaattg atgtcggtga tcatctacag ctctatcaaa gaagactaaa
     1801 gcaccgtgtt ttggacaata tggcaaagcg ggcaaatacc aatctcgttg ccaaaaggct
     1861 tccccatacc tatttagctg aagagcaacg caatcagggc agtatctaaa gcggttatcg
     1921 ctcttaactc tagaagcagc gactcctagc attaaatgca ccgcaccttg cgcttggtac
     1981 tccattaacc gaatagcttc gtctcggcgc tccttgccta caaacggagc atataaaggg
     2041 aataaggtat gctcataaat aagctgctga acagcgtaac gtcctgtttg atgtagatgt
     2101 cttgctatca cacctaaatg cgagggcaga cctaaggtag cgaccacctt gcggttgcca
     2161 tacacctcat ccaacagctg cttaggacta acaatccctt gataaacgcc tgcccgtgca
     2221 atagtgctat aaatcagctc attcgagtac ggaacaggaa agtttctcat ggctaacctg
     2281 cttgtttaaa taagctcgcc atatccacta ttaccccttt cccttttaac cgttcatgca
     2341 tggttttttc aggttggcgt tgtgaataga tataacgtaa atccgtatca ggcaagctat
     2401 cccagtcgct gggcttgact acctttatag ccgaaacctt tttactcttg gagggctttt
     2461 ctagttctga cactaccgtt tcgccttcca tcaaccactg caaaacaaga ggcagtaact
     2521 tttgtcttgt catcgttgga ttctggctaa acgctttttt aatagtggga attaacaggc
     2581 ttgaatcgta atcctctttc agcatcagat ataaatgacg ctgatcttcg gtatctaact
     2641 cttgaagggc tttttcttct ggtgtttgtt cttgtatcgc tgcgatatct agctgaagtt
     2701 ggattaaccg tttatcaatc tcgggaacga ctagatcaga ataacgagca atgcgttctg
     2761 ggatacccga gcgtaatgcc tctagcatgg ggtgcacagg ctttaactca tcttgataca
     2821 cttgccgcaa taaaccagcg gtaatacgct cattgcctaa agctagcgca cggagctgag
     2881 cgagtacaaa aagttttact acaatgtcca tcactccttg gcttagctca taccacacat
     2941 cacggacctc atccgataac agcgcatctt tgcgttgtaa aagctgtaat tgccaaagat
     3001 tatccgtaaa agcgatccac tcttgattgg gctttccacg ttgcgtttgt tgtataggat
     3061 cccagaatat agctccaaac cctgcccctc tacgtgcaga ccgcaaatca gcctcaaaaa
     3121 tctctcgtgc tttaggggta ccaatcaaca tcactggtac gccaataata ttcaccatcg
     3181 tcacaaaaaa gttcagcatc tcttgagatc cacccgaacg agagcggctt aaatgctgaa
     3241 tttcatcaat aaccaacaac cctaaagcat gtgcattggc tatttgcgac atcaaagcca
     3301 acatggtttc tataccatga cgttttaagc cataacgacg ctcatagttc gagcccaagg
     3361 ctcgatccaa cgctctgaaa aaattcaagc agatttcttt tagcgaacca ttatgcgagc
     3421 agtctatttt caaatacacc acctgctcta cattgagttc acgatggtaa atcacctgag
     3481 gatacgtggc tagaatacga tgaagagagg tcgtcttccc actaccagaa caaccaatta
     3541 acaataagct ttgtgccgta gatcgtgcct cctcaaagcg aaatgtctcc aactctcccg
     3601 tttgaacacg ctcataacca ttttgtaaat gcttttgtaa atctcctgtt ttaggatttc
     3661 tgcctacgta gccacctcga atcatgaccg aaatacgctc acttagtagc aaatgcgtac
     3721 ctaatggctg aaaatagtca tctggaatac gacaaatggt atgagctctg ataacacggg
     3781 acttttgcaa gtcagaggaa gtaagctgta aagaggattt cagtgatgca gcactattca
     3841 ctgactcttg taatggtggt aaggcctcga taaaaggatt atcacgataa gcctctaccc
     3901 ccgtatcacg ataaactgct tgaatccggg tagcactcat gactcatcct tttctggtgg
     3961 atcctgaaat aattcaggca cgtatgtggg taggctgtaa tcttcttgat catccgcttc
     4021 cactgcgttg aaaggaataa ctttagcctc atcacctgaa gagcttggct tcaaatgctc
     4081 cgcacgtttt ttacgctccg aggtcacggc ttctttttta ttagttttaa tctgcttaat
     4141 gcgtgttgat ttgggctcag tagtactggg cgttaactta ttcgctttct gaatggtttg
     4201 ctgaataaac gcctcaagct ccctgcgttt agttaactca tcctgcttcg cattggcttt
     4261 attgtgtttt tcttgtgctt gtatatccca aacctcccaa aatgagagac ctttaaactg
     4321 ccgactacgt tccgtcagat tacagcgcca aaatacacgg ctgccaactt gcggaaacaa
     4381 ataaatcgta tcaaccagca ctgggtcata agccgcttct aaatgttgag gtctagctat
     4441 atcagtgctc cgctgcaacc aaccctcacg cagaatctct gaccccgagt aatacaaacc
     4501 ccacaaatta acgccaaatg aagaaataga gacctttcgg cgaggcagta acgctactcg
     4561 caactgctct tgctccacag cccttaaact acctgtacga tgctgcatac cccattgcca
     4621 tagctggaca ggaatagacg gtaaatctgt aggaaaatca gcatctcgat cgtatttatc
     4681 catcaccaga tggttatttc tgaataagat cgtacgcaaa ataatttgtg tgaactcaaa
     4741 taccgacaga gatgcatcta acctatagtc tgtttcacca tggcttttga tccgactgcc
     4801 ctctacaatg ccaggtgcaa aggacttaaa ctcggcttgt agtgttctaa aagtgctttc
     4861 cactatgcct ttagcatcgc cacgtctagg tggagcactt tccactcgca cattaaaact
     4921 agaaactaag gcttcgacct gatgactcat taattcgcca cggtccgcta gcaacacatc
     4981 tggcaaacct acacacggcc agtctgagct actaatctca atatcatgct gggcacaaat
     5041 ggccgttttg tcagagcaag catttacaaa agcctgcatc gccaccacat aagacggatt
     5101 ttcaaagccg atataaaagc ccgtgatcat ccgactaaac acatcaatca caatgtaaag
     5161 cgttggtctt cctatgattt tttggcgatc atgatgatcc actaaataaa tatcagcaat
     5221 cgtggcatca atctcataac gactcccagg gcctaacgcc tgagaagtgg ctgtactact
     5281 taagggtcgt acgtcttttt tatatacccc tgctttaact ctagacttta agcgctgagc
     5341 tttagggtat tctcgatcat aaaaataacg aaactgacgt agtgttgggt aatcctcttg
     5401 gggaatgcga ggaaaatact gagcaaacaa gtccacaaat cgtctatagg caacggtggt
     5461 ctttgtacct ttttgattta acaggtgctt ttctatggtc aacctaaaaa ggcgttcaat
     5521 ctcgggcgtt accttggttc cttcaccctt accatattct ctggctcggc caatctttgc
     5581 tgttcctgtc gctgaacgtc tttcccctgg tgcaccgctg tttttgtagt caggaattaa
     5641 tgcattaggc gtttgaccac gctgccagta acggcgtaac aacttataaa ctgtagcctt
     5701 agtaacctta tgttcttgga ccacatgctc aacgagttcg cttctgactt tagggtcgaa
     5761 acgatcctta ctattaataa taggaagaat ttttcgataa tcctcctcgc gcttctggaa
     5821 gctgacagaa tctacagacg gctcttctaa atcaagatgt acataagggt catcagttct
     5881 aactaagcgg ccttcatcta aatactgcat tagcaacaac tcagccctag cttgtggaac
     5941 tcctttatca gcgcttattt gcatccagac aacttggcca tcctctatag ccaaaatgcg
     6001 atacggatca ttatcaaata gcacaacctc attaatttgc cacatagcgc aactcctcca
     6061 tattcactac ttggctaata cagagatctg cacacttatt tgccctgaaa gacttataaa
     6121 tattgaactt aataaaacca tttgcggtta aggctcgtat ctcactcaat gttttgccta
     6181 actccaaatc ataagcaata tcaacctgct tacagacatt gataatgttt tcatctcctt
     6241 tttcttgcag gatatgggcc aatggggata gttgtgctaa aagctccgca gaaacttctt
     6301 ctgttttcac tgaataaagc cattcaatat tttcttttac tacgggattt atttctttat
     6361 cagtaaaaat gaaccaagga atttgctttt gctgccaata gcgacgctct agttctagtt
     6421 tttctaaggt acgctcgtct tgtaaggctg ctgcaggttt gacttgaata gcaaactgct
     6481 caaaaggacc atctttgcag tccactaaaa aatcagtaga cataacctga tctacaccac
     6541 gaataacagg atgcttaata ccactatcta ttgcaatctg cctggtatca ctaggtaata
     6601 aggggaactg ctcgcgtata tctagcacgc tgctctccca ctcaagactg agaaaaacag
     6661 caagctctaa gtcagatagc aaatgatgga ctcgtcccgt cttatgagaa taaatacggt
     6721 gggaacgacc tgaagaagga acttcttgta ctgttagcca tggaatatag tctttaccat
     6781 gcccttggcc acgcccctct ttaatacgac gggcaatttg cacttcagaa aatgaagagt
     6841 ttgctttagc cataacaaaa gtccagtatg ctttttcaca gcataactgg actgatttca
     6901 gtttacaact attctgtcta gtttaagact ttattgtcat agtttagatc tattttgttc
     6961 agtttaagac tttattgtcc gcccacaccc gcttacgcag ggcatccatt tattactcaa
     7021 ccgtaaccga ttttgccagg ttacgcggct ggtcaacgtc ggtgcctttg atcagcgcga
     7081 catggtaagc cagcagctgc agcggaacgg tgtagaagat cggtgcaatc acctcttcca
     7141 catgcggcat ctcgactcta gaggatcccc gggtaccgag ctcgaattcg taatcatgtc
     7201 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     7261 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     7321 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     7381 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     7441 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     7501 acggttatcc acagaatcag gggataacgc aggaaagaac atttaaggta tactttccgc
     7561 tgcataaccc tgcttcgggg tcattatagc gattttttcg gtatatccat cctttttcgc
     7621 acgatataca ggattttgcc aaagggttcg tgtagacttt ccttggtgta tccaacggcg
     7681 tcagccgggc aggataggtg aagtaggccc acccgcgagc gggtgttcct tcttcactgt
     7741 cccttattcg cacctggcgg tgctcaacgg gaatcctgct ctgcgaggct ggccgataag
     7801 ctctgataac gcaggaaaga acatgggcag ttcaacctgt tgatagtacg tactaagctc
     7861 tcatgtttca cgtactaagc tctcaagttt aacgtactaa gctctcatgt ttaacgaact
     7921 aaaccctcat ggctaacgta ctaagctctc atggctaacg tactaagctc tcatgtttca
     7981 cgtactaagc tctcatgttt gaacaataaa attaatataa atcagcaact taaatagcct
     8041 ctaaggtttt aagttttata agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg
     8101 tttaacggtt gtggacaaca agccagggat gtaacgcact gagaagccct tagagcctct
     8161 caaagcaatt ttgagtgaca caggaacact taacggctga caaccggctg gtaacaggat
     8221 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     8281 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     8341 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     8401 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     8461 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8521 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8581 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8641 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8701 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8761 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8821 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8881 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8941 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     9001 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     9061 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     9121 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     9181 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     9241 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     9301 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     9361 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     9421 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     9481 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9541 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     9601 tgacgtctaa gaaac
//
