LOCUS       sequence-228632-        7403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn7 tagging vector pTn7xKS with dTomato expression scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7403)
  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 7403)
  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: pTn7xKS-dTomato
FEATURES             Location/Qualifiers
     source          1..7403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            230..338
                     /note="incP origin of transfer"
     mobile_element  497..662
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     primer_bind     complement(820..840)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             complement(925..1626)
                     /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(1008..1031)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1206..1225)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1475..1493
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     RBS             1634..1656
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(join(1680..1686,1687..1702,1703..1708))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    complement(1756..1803)
                     /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(1844..1849,1850..1866,1867..1872)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2061..2594
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(2724..2771)
                     /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(2817..2903)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3006..3100)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3128..3144)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3129..3145)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    3534..3555
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(3568..4650)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISIVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4612..4631
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4789..4794,4795..4810,4811..4817)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4825..4841
                     /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)."
     RBS             4981..4989
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(5412..5431)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5531..5553
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5591..5609)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5677..5781
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5782..5850,5851..6642)
                     /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(6000..6019)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6813..7401
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7302..7321
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
       61 ctttcctgcg ttatcagggg attccttaag gtatactttc cgctgcataa ccctgcttcg
      121 gggtcattat agcgattttt tcggtatatc catccttttt cgcacgatat acaggatttt
      181 gccaaagggt tcgtgtagac tttccttggt gtatccaacg gcgtcagccg ggcaggatag
      241 gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg
      301 cggtgctcaa cgggaatcct gctctgcgag gctggccgat aagctctgat accgctcgcc
      361 gcagccgaac gaccgagcgc agcgggtcag tgagcgagga agcggaagag cgcccaatac
      421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcgcccaa acataacagg
      481 aagaaaaatg ccccgctgtg ggcggacaaa atagttggga actgggaggg gtggaaatgg
      541 agtttttaag gattatttag ggaagagtga caaaatagat gggaactggg tgtagcgtcg
      601 taagctaata cgaaaattaa aaatgacaaa atagtttgga actagatttc acttatctgg
      661 ttggcctgca aggccttcgc gaggtaccgg gcccaagctt ctcgaggaat tcctgcagcc
      721 cgggggatcc acctaggccg agaaaaaaaa gcccgctcat taggcgggct agctcattat
      781 ttgtagagct catccatgcc atgtgtaatc ccagcagcag ttacaaactc aagaaggacc
      841 atgtggtcac gcttttcgtt gggatctttc gaaagggcag attgtgtcga ccagctatcg
      901 atccatgggg cccgcggccg cttacttgta cagctcgtcc atgccgtaca ggaacaggtg
      961 gtggcggccc tcggagcgct cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt
     1021 gatgtccagc ttggtgtcca cgtagtagta gccgggcagt tgcacgggct tcttggccat
     1081 gtagatggtc ttgaactcca ccaggtagtg gccgccgtcc ttcagcttca gggcctggtg
     1141 gatctcgccc ttcagcacgc cgtcgcgggg gtacaggcgc tcggtggagg cctcccagcc
     1201 catggtcttc ttctgcatta cggggccgtc gggggggaag ttggtgccgc gcatcttcac
     1261 cttgtagatc agcgtgccgt cctgcaggga ggagtcctgg gtcacggtca ccagaccgcc
     1321 gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcttgta
     1381 atcggggatg tcggcggggt gcttcacgta cgccttggag ccgtacatga actgggggga
     1441 caggatgtcc caggcgaagg gcagggggcc gcccttggtc accttcagct tggcggtctg
     1501 ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcat
     1561 ggagccctcc atgcgcacct tgaagcgcat gaactctttg atgacctcct cgcccttgct
     1621 caccatatgt atatctcctt cttaaagtta aacaaaatta tttctaggga tcctccacac
     1681 attatacgag ccgatgatta attgtcaaga atttagtgag ctcatgcatg atcgaattag
     1741 cttcaaaagc gctctgaagt tcctatactt tctagagaat aggaacttcg gaataggaac
     1801 ttcaagatcc cctgattccc tttgtcaaca gcaatggatc gaattgacat aagcctgttc
     1861 ggttcgtaaa ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac
     1921 cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt
     1981 tgtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
     2041 gatgttatgg agcagcaacg atgttacgca gcagcaacga tgttacgcag cagggcagtc
     2101 gccctaaaac aaagttaggt ggctcaagta tgggcatcat tcgcacatgt aggctcggcc
     2161 ctgaccaagt caaatccatg cgggctgctc ttgatctttt cggtcgtgag ttcggagacg
     2221 tagccaccta ctcccaacat cagccggact ccgattacct cgggaacttg ctccgtagta
     2281 agacattcat cgcgcttgct gccttcgacc aagaagcggt tgttggcgct ctcgcggctt
     2341 acgttctgcc caagtttgag cagccgcgta gtgagatcta tatctatgat ctcgcagtct
     2401 ccggcgagca ccggaggcag ggcattgcca ccgcgctcat caatctcctc aagcatgagg
     2461 ccaacgcgct tggtgcttat gtgatctacg tgcaagcaga ttacggtgac gatcccgcag
     2521 tggctctcta tacaaagttg ggcatacggg aagaagtgat gcactttgat atcgacccaa
     2581 gtaccgccac ctaacaattc gttcaagccg agatcggctt cccggccgcg gagttgttcg
     2641 gtaaattgtc acaacgccgc ggccaattcg atctagaatt ccattgagta agtttttaag
     2701 cacatcagct tcaaaagcgc tctgaagttc ctatactttc tagagaatag gaacttcgga
     2761 ataggtactt caagatcccc aattcgatcg tccgggccgc aagctcctag cggcggattt
     2821 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc
     2881 gactgagcct ttcgttttat ttgatgcctc aagctagaga gtcattaccc caggcgttta
     2941 agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat cgcagtctag
     3001 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     3061 tggagttctg aggtcattac tggatctatc aacaggagtc caagctcagc taattaagct
     3121 agcttatcga taccgtcgac ctcgaacccc acgcccctct ttaatacgac gggcaatttg
     3181 cacttcagaa aatgaagagt ttgctttagc cataacaaaa gtccagtatg ctttttcaca
     3241 gcataactgg actgatttca gtttacaact attctgtcta gtttaagact ttattgtcat
     3301 agtttagatc tattttgttc agtttaagac tttattgtcc gcccacacct gcttacgcag
     3361 ggcatccatt tattactcaa ccgtaaccga ttttgccagg ttacgcggct ggtcctctag
     3421 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     3481 tggcgaatgg cgatccgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3541 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3601 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3661 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     3721 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     3781 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     3841 tatcgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     3901 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     3961 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     4021 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     4081 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     4141 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     4201 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     4261 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     4321 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     4381 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     4441 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     4501 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttctcgcg ttttcgcaga
     4561 aacgtggctg gcctggttca ccacgcgaga aacggtctga taagagacac cggcatactc
     4621 tgcgacatcg tatagcgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     4681 gcgctatcat gccataccac gaaaggtttt gcaccattcg atgcgcctga tgcggtattt
     4741 tctccttacg catctgtgcg gtatttcaca ccgcatatcg aggagctgtt gacaattaat
     4801 catcggctcg tataatgtgt ggaattgtga gcggataaca atttcacacg gccggccaag
     4861 gagaaaggct atgaagcaca accctctggt ggtgtgtctg ctcattatct gcattacgat
     4921 tctgacattc acactcctga cccgacaaac gctctacgaa ctgcggttcc gggacggtga
     4981 taaggaggtt gctgcgctca tggcctgcac gtccaggtaa tcgcgagtcg acaggaagga
     5041 taagaagaac ataaaaaatg gcactattct ctaaaatatt aattttttat gtgattggtg
     5101 tgaacatatc ctttgtcatt atctggttta tctcacatga gaaaacacat attcgtttac
     5161 ttagtgcatt cctggtcgga ataacctggc caatgagtct gcctgtggca ttactttttt
     5221 ctctctttta ggtcgacggc gccaggagac gcgtatgaac ctggtggata tcgccattct
     5281 tatcctcaaa ctcattgttg cagcactgca actgcttgat gctgttctga aatacctgaa
     5341 gtaatactta attaagggca agcgcgggga ttttccccgc gcatttcccg ggtcagatgg
     5401 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     5461 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     5521 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcgtcacc gaaacgcgcg
     5581 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5641 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5701 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5761 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5821 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5881 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5941 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6001 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6061 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6121 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6181 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6241 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6301 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6361 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6421 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6481 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6541 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6601 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6661 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6721 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6781 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6841 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6901 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6961 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7021 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7081 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7141 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7201 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7261 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7321 tatagtcctg tcgggtttcg ccacctctga ctcgagcgtc gatttttgtg atgctcgtca
     7381 ggggggcgga gcctatggaa aaa
//
