LOCUS       sequence-221646-        7182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn7 tagging vector pTn7xTS with sfGFP expression scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7182)
  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 7182)
  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-sfGFP
FEATURES             Location/Qualifiers
     source          1..7182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            107..215
                     /note="incP origin of transfer"
     mobile_element  519..684
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     primer_bind     complement(831..851)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             complement(join(933..1646,1647..1649))
                     /codon_start=1
                     /product="enhanced blue variant of GFP"
                     /label=EBFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIKV
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(978..999)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             1657..1679
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(join(1703..1709,1710..1725,1726..1731))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    complement(1781..1828)
                     /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(1869..1874,1875..1891,1892..1897)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2086..2619
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(2749..2796)
                     /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(2857..2943)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3046..3140)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3168..3184)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3169..3185)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3599..3618)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3718..3740
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3778..3796)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3864..3968
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3969..4037,4038..4829)
                     /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(4187..4206)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(5488..6438)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      6486..6708
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tggtactatc
       61 aacaggttga actgcccatg ttctttcctg cgttatcaga gcttatcggc cagcctcgca
      121 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc
      181 cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag
      241 gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata
      301 ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaagt ataccttaag
      361 gaatcccctg ataccgctcg ccgcagccga acgaccgagc gcagcgggtc agtgagcgag
      421 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
      481 tgcagcgccc aaacataaca ggaagaaaaa tgccccgctg tgggcggaca aaatagttgg
      541 gaactgggag gggtggaaat ggagttttta aggattattt agggaagagt gacaaaatag
      601 atgggaactg ggtgtagcgt cgtaagctaa tacgaaaatt aaaaatgaca aaatagtttg
      661 gaactagatt tcacttatct ggttggcctg caaggccttc gcgaggtacc gggcccaagc
      721 ttctcgagga attcctgcag cccctaggcc gagaaaaaaa agcccgctca ttaggcgggc
      781 tagctcatta tttgtagagc tcatccatgc catgtgtaat cccagcagca gttacaaact
      841 caagaaggac catgtggtca cgcttttcgt tgggatcttt cgaaagggca gattgtgtcg
      901 accagctatc gatccatggg gcccgcggcc gctcacttgt acagctcgtc catgccgaga
      961 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
     1021 tctttgctca ggacggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg
     1081 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgacg
     1141 ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag
     1201 acgttgtggc tgttgaagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg
     1261 aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg
     1321 gcgcgggtct tgtaggtgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct
     1381 tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag
     1441 cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg
     1501 gtggtgcaga tgaacttcag ggtcagcttg ccgttggtgg catcgccctc gccctcgccg
     1561 cgcacgctga acttgtggcc gtttacgtcg ccatccagct cgaccaggat gggcaccacc
     1621 ccggtgaaca gctcctcgcc cttgctcata tgtatatctc cttcttaaag ttaaacaaaa
     1681 ttatttctag ggatcctcca cacattatac gagccgatga ttaattgtca agaattgggg
     1741 gatccactag tgagctccga attagcttca aaagcgctct gaagttccta tactttctag
     1801 agaataggaa cttcggaata ggaacttcaa gatcccctga ttccctttgt caacagcaat
     1861 ggatcgaatt gacataagcc tgttcggttc gtaaactgta atgcaagtag cgtatgcgct
     1921 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     1981 tcatggcttg ttatgactgt ttttttgtac agtctatgcc tcgggcatcc aagcagcaag
     2041 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagc
     2101 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taggtggctc aagtatgggc
     2161 atcattcgca catgtaggct cggccctgac caagtcaaat ccatgcgggc tgctcttgat
     2221 cttttcggtc gtgagttcgg agacgtagcc acctactccc aacatcagcc ggactccgat
     2281 tacctcggga acttgctccg tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa
     2341 gcggttgttg gcgctctcgc ggcttacgtt ctgcccaagt ttgagcagcc gcgtagtgag
     2401 atctatatct atgatctcgc agtctccggc gagcaccgga ggcagggcat tgccaccgcg
     2461 ctcatcaatc tcctcaagca tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa
     2521 gcagattacg gtgacgatcc cgcagtggct ctctatacaa agttgggcat acgggaagaa
     2581 gtgatgcact ttgatatcga cccaagtacc gccacctaac aattcgttca agccgagatc
     2641 ggcttcccgg ccgcggagtt gttcggtaaa ttgtcacaac gccgcggcca attcgatcta
     2701 gaattccatt gagtaagttt ttaagcacat cagcttcaaa agcgctctga agttcctata
     2761 ctttctagag aataggaact tcggaatagg tacttcaaga tccccaattc gatcgtcatg
     2821 catgatatcg tccgggccgc aagctcctag cggcggattt gtcctactca ggagagcgtt
     2881 caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat
     2941 ttgatgcctc aagctagaga gtcattaccc caggcgttta agggcaccaa taactgcctt
     3001 aaaaaaatta cgccccgccc tgccactcat cgcagtctag cttggattct caccaataaa
     3061 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     3121 tggatctatc aacaggagtc caagctcagc taattaagct agcttatcga taccgtcgac
     3181 ctcgaacccc acgcccctct ttaatacgac gggcaatttg cacttcagaa aatgaagagt
     3241 ttgctttagc cataacaaaa gtccagtatg ctttttcaca gcataactgg actgatttca
     3301 gtttacaact attctgtcta gtttaagact ttattgtcat agtttagatc tattttgttc
     3361 agtttaagac tttattgtcc gcccacaccc gcttacgcag ggcatccatt tattactcaa
     3421 ccgtaaccga ttttgccagg ttacgcggct ggtcctctag ctggcgtaat agcgaagagg
     3481 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     3541 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     3601 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     3661 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3721 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     3781 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     3841 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3901 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3961 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4021 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4081 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4141 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4201 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4261 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4321 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4381 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4441 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4501 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4561 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4621 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4681 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4741 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4801 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4861 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4921 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4981 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5041 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5101 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     5161 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5221 tcctgttacc agccggttgt cagccgttaa gtgttcctgt gtcactcaaa attgctttga
     5281 gaggctctaa gggcttctca gtgcgttaca tccctggctt gttgtccaca accgttaaac
     5341 cttaaaagct ttaaaagcct tatatattct tttttttctt acatgggtat ggacagtttt
     5401 ccctttgata tgtaacggtg aacagttgtt ctacttttgt ttgttagtct tgatgcttca
     5461 ctgatagata caagagccat aagaacctca gatccttccg tatttagcca gtatgttctc
     5521 tagtgtggtt cgttgttttt gcgtgagcca tgagaacgaa ccattgagat catacttact
     5581 ttgcatgtca ctcaaaaatt ttgcctcaaa actggtgagc tgaatttttg cagttaaagc
     5641 atcgtgtagt gtttttctta gtccgttacg taggtaggaa tctgatgtaa tggttgttgg
     5701 tattttgtca ccattcattt ttatctggtt gttctcaagt tcggttacga gatccatttg
     5761 tctatctagt tcaacttgga aaatcaacgt atcagtcggg cggcctcgct tatcaaccac
     5821 caatttcata ttgctgtaag tgtttaaatc tttacttatt ggtttcaaaa cccattggtt
     5881 aagcctttta aactcatggt agttattttc aagcattaac atgaacttaa attcatcaag
     5941 gctaatctct atatttgcct tgtgagtttt cttttgtgtt agttctttta ataaccactc
     6001 ataaatcctc atagagtatt tgttttcaaa agacttaaca tgttccagat tatattttat
     6061 gaattttttt aactggaaaa gataaggcaa tatctcttca ctaaaaacta attctaattt
     6121 ttcgcttgag aacttggcat agtttgtcca ctggaaaatc tcaaagcctt taaccaaagg
     6181 attcctgatt tccacagttc tcgtcatcag ctctctggtt gctttagcta atacaccata
     6241 agcattttcc ctactgatgt tcatcatctg agcgtattgg ttataagtga acgataccgt
     6301 ccgttctttc cttgtagggt tttcaatcgt ggggttgagt agtgccacac agcataaaat
     6361 tagcttggtt tcatgctccg ttaagtcata gcgactaatc gctagttcat ttgctttgaa
     6421 aacaactaat tcagacatac atctcaattg gtctaggtga ttttaatcac tataccaatt
     6481 gagatgggct agtcaatgat aattactagt ccttttcctt tgagttgtgg gtatctgtaa
     6541 attctgctag acctttgctg gaaaacttgt aaattctgct agaccctctg taaattccgc
     6601 tagacctttg tgtgtttttt ttgtttatat tcaagtggtt ataatttata gaataaagaa
     6661 agaataaaaa aagataaaaa gaatagatcc cagccctgtg tataactcac tactttagtc
     6721 agttccgcag tattacaaaa ggatgtcgca aacgctgttt gctcctctac aaaacagacc
     6781 ttaaaaccct aaaggcttaa gtagcaccct cgcaagctcg gttgcggccg caatcgggca
     6841 aatcgctgaa tattcctttt gtctccgacc atcaggcacc tgagtcgctg tctttttcgt
     6901 gacattcagt tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg gcactacagg
     6961 cgccttttat ggattcatgc aaggaaacta cccataatac aagaaaagcc cgtcacgggc
     7021 ttctcagggc gttttatggc gggtctgcta tgtggtgcta tctgactttt tgctgttcag
     7081 cagttcctgc cctctgattt tccagtctga ccacttcgga ttatcccgtg acaggtcatt
     7141 cagactggct aatgcaccca gtaaggcagc ggtatcatca ac
//
