LOCUS       sequence-230523-        7338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn7 tagging vector pTn7xTS with mPlum expression scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7338)
  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 7338)
  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-mPlum
FEATURES             Location/Qualifiers
     source          1..7338
                     /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     793..812
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1023..1043)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             complement(1125..1805)
                     /codon_start=1
                     /product="far-red monomeric derivative of DsRed fluorescent
                     protein (Wang et al., 2004)"
                     /label=mPlum
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKEHMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     RLKVTKGGPLPFAWDILSPQIMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGEFIYKVKVRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEMK
                     MRLRLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRH
                     STGA"
     primer_bind     complement(1187..1210)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1385..1404)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     RBS             1813..1835
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(join(1859..1865,1866..1881,1882..1887))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    complement(1937..1984)
                     /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(2025..2030,2031..2047,2048..2053)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2242..2775
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(2905..2952)
                     /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(3013..3099)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3202..3296)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3324..3340)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3325..3341)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3755..3774)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3874..3896
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3934..3952)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4020..4124
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4125..4193,4194..4985)
                     /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(4343..4362)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(5644..6594)
                     /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      6642..6864
                     /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 ccccggtaaa ccgaaaggca ggaacaggag agcgcacggg
      781 ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg
      841 atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt
      901 tgccgcggcc ctctcacttc cctgttaagt atctcctagg ccgagaaaaa aaagcccgct
      961 cattaggcgg gctagctcat tatttgtaga gctcatccat gccatgtgta atcccagcag
     1021 cagttacaaa ctcaagaagg accatgtggt cacgcttttc gttgggatct ttcgaaaggg
     1081 cagattgtgt cgaccagcta tcgatccatg gggcccgcgg ccgcctaggc gccggtggag
     1141 tggcggccct cggcgcgctc gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg
     1201 atgtccagct tgatgtcggt cttgtaggcg ccgggcagct gcacgggctt cttggccatg
     1261 taggtggtct tgacctcggc gtcgtagtgg ccgccgtcct tcagcctcag cctcatcttc
     1321 atctcgccct tcagggcgcc gtcctcgggg tacatccgct cggaggaggc ctcccagccc
     1381 atggtcttct tctgcattac ggggccgtcg gaggggaagt tggtgccgcg caccttcacc
     1441 ttgtagatga actcgccgtc ctgcagggag gagtcctggg tcacggtcac cacgccgccg
     1501 tcctcgaagt tcatcacgcg ctcccacttg aagccctcgg ggaaggacag cttcaagtag
     1561 tcggggatgt cggcggggtg cttcacgtag gccttggagc cgtacatgat ctgaggggac
     1621 aggatgtccc aggcgaaggg cagggggcca cccttggtca ccttcagcct ggcggtctgg
     1681 gtgccctcgt aggggcggcc ctcgccctcg ccctcgatct cgaactcgtg gccgttcacg
     1741 gagccctcca tgtgctcctt gaagcgcatg aactccttga tgacctcctc gcccttgctc
     1801 accatatgta tatctccttc ttaaagttaa acaaaattat ttctagggat cctccacaca
     1861 ttatacgagc cgatgattaa ttgtcaagaa ttgggggatc cactagtgag ctccgaatta
     1921 gcttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggaa
     1981 cttcaagatc ccctgattcc ctttgtcaac agcaatggat cgaattgaca taagcctgtt
     2041 cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga
     2101 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt
     2161 ttgtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
     2221 tgatgttatg gagcagcaac gatgttacgc agcagcaacg atgttacgca gcagggcagt
     2281 cgccctaaaa caaagttagg tggctcaagt atgggcatca ttcgcacatg taggctcggc
     2341 cctgaccaag tcaaatccat gcgggctgct cttgatcttt tcggtcgtga gttcggagac
     2401 gtagccacct actcccaaca tcagccggac tccgattacc tcgggaactt gctccgtagt
     2461 aagacattca tcgcgcttgc tgccttcgac caagaagcgg ttgttggcgc tctcgcggct
     2521 tacgttctgc ccaagtttga gcagccgcgt agtgagatct atatctatga tctcgcagtc
     2581 tccggcgagc accggaggca gggcattgcc accgcgctca tcaatctcct caagcatgag
     2641 gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag attacggtga cgatcccgca
     2701 gtggctctct atacaaagtt gggcatacgg gaagaagtga tgcactttga tatcgaccca
     2761 agtaccgcca cctaacaatt cgttcaagcc gagatcggct tcccggccgc ggagttgttc
     2821 ggtaaattgt cacaacgccg cggccaattc gatctagaat tccattgagt aagtttttaa
     2881 gcacatcagc ttcaaaagcg ctctgaagtt cctatacttt ctagagaata ggaacttcgg
     2941 aataggtact tcaagatccc caattcgatc gtcatgcatg atatcgtccg ggccgcaagc
     3001 tcctagcggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     3061 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctcaagc tagagagtca
     3121 ttaccccagg cgtttaaggg caccaataac tgccttaaaa aaattacgcc ccgccctgcc
     3181 actcatcgca gtctagcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt
     3241 tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag
     3301 ctcagctaat taagctagct tatcgatacc gtcgacctcg aaccccacgc ccctctttaa
     3361 tacgacgggc aatttgcact tcagaaaatg aagagtttgc tttagccata acaaaagtcc
     3421 agtatgcttt ttcacagcat aactggactg atttcagttt acaactattc tgtctagttt
     3481 aagactttat tgtcatagtt tagatctatt ttgttcagtt taagacttta ttgtccgcct
     3541 acacccgctt acgcagggca tccatttatt actcaaccgt aaccgatttt gccaggttac
     3601 gcggctggtc ctctagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     3661 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     3721 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3781 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3841 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3901 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     3961 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtac
     4021 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4081 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4141 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4201 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4261 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4321 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4381 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4441 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4501 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4561 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4621 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     4681 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4741 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4801 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4861 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4921 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4981 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5041 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5101 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5161 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5221 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5281 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5341 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagcc ggttgtcagc
     5401 cgttaagtgt tcctgtgtca ctcaaaattg ctttgagagg ctctaagggc ttctcagtgc
     5461 gttacatccc tggcttgttg tccacaaccg ttaaacctta aaagctttaa aagccttata
     5521 tattcttttt tttcttacat gggtatggac agttttccct ttgatatgta acggtgaaca
     5581 gttgttctac ttttgtttgt tagtcttgat gcttcactga tagatacaag agccataaga
     5641 acctcagatc cttccgtatt tagccagtat gttctctagt gtggttcgtt gtttttgcgt
     5701 gagccatgag aacgaaccat tgagatcata cttactttgc atgtcactca aaaattttgc
     5761 ctcaaaactg gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc
     5821 gttatgtagg taggaatctg atgtaatggt tgttggtatt ttgtcaccat tcatttttat
     5881 ctggttgttc tcaagttcgg ttacgagatc catttgtcta tctagttcaa cttggaaaat
     5941 caacgtatca gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt
     6001 taaatcttta cttattggtt tcaaaaccca ttggttaagc cttttaaact catggtagtt
     6061 attttcaagc attaacatga acttaaattc atcaaggcta atctctatat ttgccttgtg
     6121 agttttcttt tgtgttagtt cttttaataa ccactcataa atcctcatag agtatttgtt
     6181 ttcaaaagac ttaacatgtt ccagattata ttttatgaat ttttttaact ggaaaagata
     6241 aggcaatatc tcttcactaa aaactaattc taatttttcg cttgagaact tggcatagtt
     6301 tgtccactgg aaaatctcaa agcctttaac caaaggattc ctgatttcca cagttctcgt
     6361 catcagctct ctggttgctt tagctaatac accataagca ttttccctac tgatgttcat
     6421 catctgaacg tattggttat aagtgaacga taccgtccgt tctttccttg tagggttttc
     6481 aatcgtgggg ttgagtagtg ccacacagca taaaattagc ttggtttcat gctccgttaa
     6541 gtcatagcga ctaatcgcta gttcatttgc tttgaaaaca actaattcag acatacatct
     6601 caattggtct aggtgatttt aatcactata ccaattgaga tgggctagtc aatgataatt
     6661 actagtcctt ttcctttgag ttgtgggtat ctgtaaattc tgctagacct ttgctggaaa
     6721 acttgtaaat tctgctagac cctctgtaaa ttccgctaga cctttgtgtg ttttttttgt
     6781 ttatattcaa gtggttataa tttatagaat aaagaaagaa taaaaaaaga taaaaagaat
     6841 agatcccagc cctgtgtata actcactact ttagtcagtt ccgcagtatt acaaaaggat
     6901 gtcgcaaacg ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag gcttaagtag
     6961 caccctcgca agctcggttg cggccgcaat cgggcaaatc gctgaatatt ccttttgtct
     7021 ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg
     7081 gctctggcag tgaatggggg taaatggcac tacaggcgcc ttttatggat tcatgcaagg
     7141 aaactaccca taatacaaga aaagcccgtc acgggcttct cagggcgttt tatggcgggt
     7201 ctgctatgtg gtgctatctg actttttgct gttcagcagt tcctgccctc tgattttcca
     7261 gtctgaccac ttcggattat cccgtgacag gtcattcaga ctggctaatg cacccagtaa
     7321 ggcagcggta tcatcaac
//
