LOCUS       sequence-230519-        7169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn7 tagging vector pTn7xTS with dTomato expression scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7169)
  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 7169)
  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-dTomato
FEATURES             Location/Qualifiers
     source          1..7169
                     /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)"
     primer_bind     complement(830..850)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             complement(935..1636)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(1018..1041)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1216..1235)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1485..1503
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     RBS             1644..1666
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(join(1690..1696,1697..1712,1713..1718))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    complement(1768..1815)
                     /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(1856..1861,1862..1878,1879..1884)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2073..2606
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(2736..2783)
                     /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(2844..2930)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3033..3127)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3155..3171)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3156..3172)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3586..3605)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3705..3727
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3765..3783)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3851..3955
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3956..4024,4025..4816)
                     /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(4174..4193)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(5475..6425)
                     /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      6473..6695
                     /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 ccctaggccg agaaaaaaaa gcccgctcat taggcgggct
      781 agctcattat ttgtagagct catccatgcc atgtgtaatc ccagcagcag ttacaaactc
      841 aagaaggacc atgtggtcac gcttttcgtt gggatctttc gaaagggcag attgtgtcga
      901 ccagctatcg atccatgggg cccgcggccg cttacttgta cagctcgtcc atgccgtaca
      961 ggaacaggtg gtggcggccc tcggagcgct cgtactgttc cacgatggtg tagtcctcgt
     1021 tgtgggaggt gatgtccagc ttggtgtcca cgtagtagta gccgggcagt tgcacgggct
     1081 tcttggccat gtagatggtc ttgaactcca ccaggtagtg gccgccgtcc ttcagcttca
     1141 gggcctggtg gatctcgccc ttcagcacgc cgtcgcgggg gtacaggcgc tcggtggagg
     1201 cctcccagcc catggtcttc ttctgcatta cggggccgtc gggggggaag ttggtgccgc
     1261 gcatcttcac cttgtagatc agcgtgccgt cctgcaggga ggagtcctgg gtcacggtca
     1321 ccagaccgcc gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca
     1381 gcttcttgta atcggggatg tcggcggggt gcttcacgta cgccttggag ccgtacatga
     1441 actgggggga caggatgtcc caggcgaagg gcagggggcc gcccttggtc accttcagct
     1501 tggcggtctg ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt
     1561 ggccgttcat ggagccctcc atgcgcacct tgaagcgcat gaactctttg atgacctcct
     1621 cgcccttgct caccatatgt atatctcctt cttaaagtta aacaaaatta tttctaggga
     1681 tcctccacac attatacgag ccgatgatta attgtcaaga attgggggat ccactagtga
     1741 gctccgaatt agcttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt
     1801 cggaatagga acttcaagat cccctgattc cctttgtcaa cagcaatgga tcgaattgac
     1861 ataagcctgt tcggttcgta aactgtaatg caagtagcgt atgcgctcac gcaactggtc
     1921 cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta
     1981 tgactgtttt tttgtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt
     2041 gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagcaac gatgttacgc
     2101 agcagggcag tcgccctaaa acaaagttag gtggctcaag tatgggcatc attcgcacat
     2161 gtaggctcgg ccctgaccaa gtcaaatcca tgcgggctgc tcttgatctt ttcggtcgtg
     2221 agttcggaga cgtagccacc tactcccaac atcagccgga ctccgattac ctcgggaact
     2281 tgctccgtag taagacattc atcgcgcttg ctgccttcga ccaagaagcg gttgttggcg
     2341 ctctcgcggc ttacgttctg cccaagtttg agcagccgcg tagtgagatc tatatctatg
     2401 atctcgcagt ctccggcgag caccggaggc agggcattgc caccgcgctc atcaatctcc
     2461 tcaagcatga ggccaacgcg cttggtgctt atgtgatcta cgtgcaagca gattacggtg
     2521 acgatcccgc agtggctctc tatacaaagt tgggcatacg ggaagaagtg atgcactttg
     2581 atatcgaccc aagtaccgcc acctaacaat tcgttcaagc cgagatcggc ttcccggccg
     2641 cggagttgtt cggtaaattg tcacaacgcc gcggccaatt cgatctagaa ttccattgag
     2701 taagttttta agcacatcag cttcaaaagc gctctgaagt tcctatactt tctagagaat
     2761 aggaacttcg gaataggtac ttcaagatcc ccaattcgat cgtcatgcat gatatcgtcc
     2821 gggccgcaag ctcctagcgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
     2881 cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctcaag
     2941 ctagagagtc attaccccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc
     3001 cccgccctgc cactcatcgc agtctagctt ggattctcac caataaaaaa cgcccggcgg
     3061 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac
     3121 aggagtccaa gctcagctaa ttaagctagc ttatcgatac cgtcgacctc gaaccccacg
     3181 cccctcttta atacgacggg caatttgcac ttcagaaaat gaagagtttg ctttagccat
     3241 aacaaaagtc cagtatgctt tttcacagca taactggact gatttcagtt tacaactatt
     3301 ctgtctagtt taagacttta ttgtcatagt ttagatctat tttgttcagt ttaagacttt
     3361 attgtccgcc tacacccgct tacgcagggc atccatttat tactcaaccg taaccgattt
     3421 tgccaggtta cgcggctggt cctctagctg gcgtaatagc gaagaggccc gcaccgatcg
     3481 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     3541 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     3601 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3661 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3721 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3781 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3841 gggaaatgta cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3901 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3961 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4021 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4081 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4141 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4201 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4261 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4321 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4381 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4441 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4501 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4561 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4621 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4681 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4741 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4801 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4861 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4921 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4981 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5041 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5101 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5161 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagc
     5221 cggttgtcag ccgttaagtg ttcctgtgtc actcaaaatt gctttgagag gctctaaggg
     5281 cttctcagtg cgttacatcc ctggcttgtt gtccacaacc gttaaacctt aaaagcttta
     5341 aaagccttat atattctttt ttttcttaca tgggtatgga cagttttccc tttgatatgt
     5401 aacggtgaac agttgttcta cttttgtttg ttagtcttga tgcttcactg atagatacaa
     5461 gagccataag aacctcagat ccttccgtat ttagccagta tgttctctag tgtggttcgt
     5521 tgtttttgcg tgagccatga gaacgaacca ttgagatcat acttactttg catgtcactc
     5581 aaaaattttg cctcaaaact ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt
     5641 tttcttagtc cgttatgtag gtaggaatct gatgtaatgg ttgttggtat tttgtcacca
     5701 ttcattttta tctggttgtt ctcaagttcg gttacgagat ccatttgtct atctagttca
     5761 acttggaaaa tcaacgtatc agtcgggcgg cctcgcttat caaccaccaa tttcatattg
     5821 ctgtaagtgt ttaaatcttt acttattggt ttcaaaaccc attggttaag ccttttaaac
     5881 tcatggtagt tattttcaag cattaacatg aacttaaatt catcaaggct aatctctata
     5941 tttgccttgt gagttttctt ttgtgttagt tcttttaata accactcata aatcctcata
     6001 gagtatttgt tttcaaaaga cttaacatgt tccagattat attttatgaa tttttttaac
     6061 tggaaaagat aaggcaatat ctcttcacta aaaactaatt ctaatttttc gcttgagaac
     6121 ttggcatagt ttgtccactg gaaaatctca aagcctttaa ccaaaggatt cctgatttcc
     6181 acagttctcg tcatcagctc tctggttgct ttagctaata caccataagc attttcccta
     6241 ctgatgttca tcatctgaac gtattggtta taagtgaacg ataccgtccg ttctttcctt
     6301 gtagggtttt caatcgtggg gttgagtagt gccacacagc ataaaattag cttggtttca
     6361 tgctccgtta agtcatagcg actaatcgct agttcatttg ctttgaaaac aactaattca
     6421 gacatacatc tcaattggtc taggtgattt taatcactat accaattgag atgggctagt
     6481 caatgataat tactagtcct tttcctttga gttgtgggta tctgtaaatt ctgctagacc
     6541 tttgctggaa aacttgtaaa ttctgctaga ccctctgtaa attccgctag acctttgtgt
     6601 gttttttttg tttatattca agtggttata atttatagaa taaagaaaga ataaaaaaag
     6661 ataaaaagaa tagatcccag ccctgtgtat aactcactac tttagtcagt tccgcagtat
     6721 tacaaaagga tgtcgcaaac gctgtttgct cctctacaaa acagacctta aaaccctaaa
     6781 ggcttaagta gcaccctcgc aagctcggtt gcggccgcaa tcgggcaaat cgctgaatat
     6841 tccttttgtc tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg
     6901 ctgcgctcac ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga
     6961 ttcatgcaag gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt
     7021 ttatggcggg tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct
     7081 ctgattttcc agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat
     7141 gcacccagta aggcagcggt atcatcaac
//
