LOCUS       sequence-230512-        6188 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn7 tagging vector with temperature-sensitive origin of replication.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6188)
  AUTHORS   Wiles TJ, Wall ES, Schlomann BH, Hay EA, Parthasarathy R, Guillemin 
            K
  TITLE     Modernized Tools for Streamlined Genetic Manipulation and 
            Comparative Study of Wild and Diverse Proteobacterial Lineages.
  JOURNAL   MBio. 2018 Oct 9;9(5). pii: mBio.01877-18. doi: 
            10.1128/mBio.01877-18.
  PUBMED    30301859
REFERENCE   2  (bases 1 to 6188)
  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: "MBio."; 
            date: "2018-10-9"; volume: "9(5). pii: mBio.01877-18. doi"; pages: "
            10.1128/mBio.01877-18"
COMMENT     Sequence Label: pTn7xTS
FEATURES             Location/Qualifiers
     source          1..6188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            106..214
                     /note="incP origin of transfer"
     mobile_element  518..683
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     protein_bind    complement(787..834)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        join(875..880,881..897,898..903)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1092..1625
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(1755..1802)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     terminator      complement(1863..1949)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2052..2146)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2174..2190)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2175..2191)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2605..2624)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2724..2746
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2784..2802)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2870..2974
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2975..3043,3044..3835)
                     /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     complement(3193..3212)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(4494..5444)
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      5492..5714
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgtactatca
       61 acaggttgaa ctgcccatgt tctttcctgc gttatcagag cttatcggcc agcctcgcag
      121 agcaggattc ccgttgagca ccgccaggtg cgaataaggg acagtgaaga aggaacaccc
      181 gctcgcgggt gggcctactt cacctatcct gcccggctga cgccgttgga tacaccaagg
      241 aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac
      301 cgaaaaaatc gctataatga ccccgaagca gggttatgca gcggaaagta taccttaagg
      361 aatcccctga taccgctcgc cgcagccgaa cgaccgagcg cagcgggtca gtgagcgagg
      421 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      481 gcagcgccca aacataacag gaagaaaaat gccccgctgt gggcggacaa aatagttggg
      541 aactgggagg ggtggaaatg gagtttttaa ggattattta gggaagagtg acaaaataga
      601 tgggaactgg gtgtagcgtc gtaagctaat acgaaaatta aaaatgacaa aatagtttgg
      661 aactagattt cacttatctg gttggcctgc aaggccttcg cgaggtaccg ggcccaagct
      721 tctcgaggaa ttcctgcagc ccgggggatc cactagtgag ctccgaatta gcttcaaaag
      781 cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggaa cttcaagatc
      841 ccctgattcc ctttgtcaac agcaatggat cgaattgaca taagcctgtt cggttcgtaa
      901 actgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag
      961 cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttgtacagtc
     1021 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     1081 gagcagcaac gatgttacgc agcagcaacg atgttacgca gcagggcagt cgccctaaaa
     1141 caaagttagg tggctcaagt atgggcatca ttcgcacatg taggctcggc cctgaccaag
     1201 tcaaatccat gcgggctgct cttgatcttt tcggtcgtga gttcggagac gtagccacct
     1261 actcccaaca tcagccggac tccgattacc tcgggaactt gctccgtagt aagacattca
     1321 tcgcgcttgc tgccttcgac caagaagcgg ttgttggcgc tctcgcggct tacgttctgc
     1381 ccaagtttga gcagccgcgt agtgagatct atatctatga tctcgcagtc tccggcgagc
     1441 accggaggca gggcattgcc accgcgctca tcaatctcct caagcatgag gccaacgcgc
     1501 ttggtgctta tgtgatctac gtgcaagcag attacggtga cgatcccgca gtggctctct
     1561 atacaaagtt gggcatacgg gaagaagtga tgcactttga tatcgaccca agtaccgcca
     1621 cctaacaatt cgttcaagcc gagatcggct tcccggccgc ggagttgttc ggtaaattgt
     1681 cacaacgccg cggccaattc gatctagaat tccattgagt aagtttttaa gcacatcagc
     1741 ttcaaaagcg ctctgaagtt cctatacttt ctagagaata ggaacttcgg aataggtact
     1801 tcaagatccc caattcgatc gtcatgcatg atatcgtccg ggccgcaagc tcctagcggc
     1861 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     1921 tctttcgact gagcctttcg ttttatttga tgcctcaagc tagagagtca ttaccccagg
     1981 cgtttaaggg caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca
     2041 gtctagcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa
     2101 tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag ctcagctaat
     2161 taagctagct tatcgatacc gtcgacctcg aaccccacgc ccctctttaa tacgacgggc
     2221 aatttgcact tcagaaaatg aagagtttgc tttagccata acaaaagtcc agtatgcttt
     2281 ttcacagcat aactggactg atttcagttt acaactattc tgtctagttt aagactttat
     2341 tgtcatagtt tagatctatt ttgttcagtt taagacttta ttgtccgcct acacccgctt
     2401 acgcagggca tccatttatt actcaaccgt aaccgatttt gccaggttac gcggctggtc
     2461 ctctagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     2521 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2581 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     2641 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     2701 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     2761 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     2821 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtac gcggaacccc
     2881 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2941 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3001 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3061 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3121 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3181 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3241 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3301 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3361 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3421 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3481 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3541 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3601 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3661 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3721 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3781 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3841 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3901 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3961 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4021 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4081 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4141 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4201 accgcctaca tacctcgctc tgctaatcct gttaccagcc ggttgtcagc cgttaagtgt
     4261 tcctgtgtca ctcaaaattg ctttgagagg ctctaagggc ttctcagtgc gttacatccc
     4321 tggcttgttg tccacaaccg ttaaacctta aaagctttaa aagccttata tattcttttt
     4381 tttcttacat gggtatggac agttttccct ttgatatgta acggtgaaca gttgttctac
     4441 ttttgtttgt tagtcttgat gcttcactga tagatacaag agccataaga acctcagatc
     4501 cttccgtatt tagccagtat gttctctagt gtggttcgtt gtttttgcgt gagccatgag
     4561 aacgaaccat tgagatcata cttactttgc atgtcactca aaaattttgc ctcaaaactg
     4621 gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc gttatgtagg
     4681 taggaatctg atgtaatggt tgttggtatt ttgtcaccat tcatttttat ctggttgttc
     4741 tcaagttcgg ttacgagatc catttgtcta tctagttcaa cttggaaaat caacgtatca
     4801 gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt taaatcttta
     4861 cttattggtt tcaaaaccca ttggttaagc cttttaaact catggtagtt attttcaagc
     4921 attaacatga acttaaattc atcaaggcta atctctatat ttgccttgtg agttttcttt
     4981 tgtgttagtt cttttaataa ccactcataa atcctcatag agtatttgtt ttcaaaagac
     5041 ttaacatgtt ccagattata ttttatgaat ttttttaact ggaaaagata aggcaatatc
     5101 tcttcactaa aaactaattc taatttttcg cttgagaact tggcatagtt tgtccactgg
     5161 aaaatctcaa agcctttaac caaaggattc ctgatttcca cagttctcgt catcagctct
     5221 ctggttgctt tagctaatac accataagca ttttccctac tgatgttcat catctgaacg
     5281 tattggttat aagtgaacga taccgtccgt tctttccttg tagggttttc aatcgtgggg
     5341 ttgagtagtg ccacacagca taaaattagc ttggtttcat gctccgttaa gtcatagcga
     5401 ctaatcgcta gttcatttgc tttgaaaaca actaattcag acatacatct caattggtct
     5461 aggtgatttt aatcactata ccaattgaga tgggctagtc aatgataatt actagtcctt
     5521 ttcctttgag ttgtgggtat ctgtaaattc tgctagacct ttgctggaaa acttgtaaat
     5581 tctgctagac cctctgtaaa ttccgctaga cctttgtgtg ttttttttgt ttatattcaa
     5641 gtggttataa tttatagaat aaagaaagaa taaaaaaaga taaaaagaat agatcccagc
     5701 cctgtgtata actcactact ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg
     5761 ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag gcttaagtag caccctcgca
     5821 agctcggttg cggccgcaat cgggcaaatc gctgaatatt ccttttgtct ccgaccatca
     5881 ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag
     5941 tgaatggggg taaatggcac tacaggcgcc ttttatggat tcatgcaagg aaactaccca
     6001 taatacaaga aaagcccgtc acgggcttct cagggcgttt tatggcgggt ctgctatgtg
     6061 gtgctatctg actttttgct gttcagcagt tcctgccctc tgattttcca gtctgaccac
     6121 ttcggattat cccgtgacag gtcattcaga ctggctaatg cacccagtaa ggcagcggta
     6181 tcatcaac
//
