LOCUS       sequence-221656-        7408 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 7408)
  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 7408)
  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..7408
                     /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(823..843)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             complement(928..1629)
                     /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(1011..1034)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1209..1228)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1478..1496
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     RBS             1637..1659
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(join(1683..1689,1690..1705,1706..1711))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    complement(1760..1807)
                     /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(1848..1853,1854..1870,1871..1876)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2065..2598
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(2728..2775)
                     /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(2821..2907)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3010..3104)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3132..3148)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3133..3149)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    3538..3559
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(3572..4654)
                     /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     4616..4635
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4793..4798,4799..4814,4815..4821)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4829..4845
                     /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             4985..4993
                     /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(5417..5436)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5536..5558
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5596..5614)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5682..5786
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5787..5855,5856..6647)
                     /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(6005..6024)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6818..7406
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7307..7326
                     /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 actagctagg ccgagaaaaa aaagcccgct cattaggcgg gctagctcat
      781 tatttgtaga gctcatccat gccatgtgta atcccagcag cagttacaaa ctcaagaagg
      841 accatgtggt cacgcttttc gttgggatct ttcgaaaggg cagattgtgt cgaccagcta
      901 tcgatccatg gggcccgcgg ccgcttactt gtacagctcg tccatgccgt acaggaacag
      961 gtggtggcgg ccctcggagc gctcgtactg ttccacgatg gtgtagtcct cgttgtggga
     1021 ggtgatgtcc agcttggtgt ccacgtagta gtagccgggc agttgcacgg gcttcttggc
     1081 catgtagatg gtcttgaact ccaccaggta gtggccgccg tccttcagct tcagggcctg
     1141 gtggatctcg cccttcagca cgccgtcgcg ggggtacagg cgctcggtgg aggcctccca
     1201 gcccatggtc ttcttctgca ttacggggcc gtcggggggg aagttggtgc cgcgcatctt
     1261 caccttgtag atcagcgtgc cgtcctgcag ggaggagtcc tgggtcacgg tcaccagacc
     1321 gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg acagcttctt
     1381 gtaatcgggg atgtcggcgg ggtgcttcac gtacgccttg gagccgtaca tgaactgggg
     1441 ggacaggatg tcccaggcga agggcagggg gccgcccttg gtcaccttca gcttggcggt
     1501 ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact cgtggccgtt
     1561 catggagccc tccatgcgca ccttgaagcg catgaactct ttgatgacct cctcgccctt
     1621 gctcaccata tgtatatctc cttcttaaag ttaaacaaaa ttatttctag ggatcctcca
     1681 cacattatac gagccgatga ttaattgtca agaattctag tgagctcatg catgatcgaa
     1741 ttagcttcaa aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag
     1801 gaacttcaag atcccctgat tccctttgtc aacagcaatg gatcgaattg acataagcct
     1861 gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct
     1921 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt
     1981 tttttgtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat
     2041 gtttgatgtt atggagcagc aacgatgtta cgcagcagca acgatgttac gcagcagggc
     2101 agtcgcccta aaacaaagtt aggtggctca agtatgggca tcattcgcac atgtaggctc
     2161 ggccctgacc aagtcaaatc catgcgggct gctcttgatc ttttcggtcg tgagttcgga
     2221 gacgtagcca cctactccca acatcagccg gactccgatt acctcgggaa cttgctccgt
     2281 agtaagacat tcatcgcgct tgctgccttc gaccaagaag cggttgttgg cgctctcgcg
     2341 gcttacgttc tgcccaggtt tgagcagccg cgtagtgaga tctatatcta tgatctcgca
     2401 gtctccggcg agcaccggag gcagggcatt gccaccgcgc tcatcaatct cctcaagcat
     2461 gaggccaacg cgcttggtgc ttatgtgatc tacgtgcaag cagattacgg tgacgatccc
     2521 gcagtggctc tctatacaaa gttgggcata cgggaagaag tgatgcactt tgatatcgac
     2581 ccaagtaccg ccacctaaca attcgttcaa gccgagatcg gcttcccggc cgcggagttg
     2641 ttcggtaaat tgtcacaacg ccgcggccaa ttcgatctag aattccattg agtaagtttt
     2701 taagcacatc agcttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt
     2761 cggaatagga acttcaagat ccccaattcg atcgtccggg ccgcaagctc ctagcggcgg
     2821 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     2881 tttcgactga gcctttcgtt ttatttgatg cctcaagcta gagagtcatt accccaggcg
     2941 tttaagggca ccaataactg ccttaaaaaa attacgcccc gccctgccac tcatcgcagt
     3001 ctagcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc
     3061 cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagct cagctaatta
     3121 agctagctta tcgataccgt cgacctcgaa ccccacgccc ctctttaata cgacgggcaa
     3181 tttgcacttc agaaaatgaa gagtttgctt tagccataac aaaagtccag tatgcttttt
     3241 cacagcataa ctggactgat ttcagtttac aactattctg tctagtttaa gactttattg
     3301 tcatagttta gatctatttt gttcagttta agactttatt gtccgcccac acccgcttac
     3361 gcagggcatc catttattac tcaaccgtaa ccgattttgc caggttacgc ggctggtcct
     3421 ctagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     3481 tgaatggcga atggcgatcc gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3541 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3601 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3661 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     3721 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     3781 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     3841 ccactatcga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     3901 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     3961 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     4021 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     4081 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     4141 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     4201 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     4261 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     4321 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     4381 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     4441 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     4501 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttct cgcgttttcg
     4561 cagaaacgtg gctggcctgg ttcaccacgc gagaaacggt ctgataagag acaccggcat
     4621 actctgcgac atcgtatagc gttactggtt tcacattcac caccctgaat tgactctctt
     4681 ccgggcgcta tcatgccata ccacgaaagg ttttgcacca ttcgatgcgc ctgatgcggt
     4741 attttctcct tacgcatctg tgcggtattt cacaccgcat atcgaggagc tgttgacaat
     4801 taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacggccggc
     4861 caaggagaaa ggctatgaag cacaaccctc tggtggtgtg tctgctcatt atctgcatta
     4921 cgattctgac attcacactc ctgacccgac aaacgctcta cgaactgcgg ttccgggacg
     4981 gtgataagga ggttgctgcg ctcatggcct gcacgtccag gtaatcgcga gtcgacagga
     5041 aggataagaa gaacataaaa aatggcacta ttctctaaaa tattaatttt ttatgtgatt
     5101 ggtgtgaaca tatcctttgt cattatctgg tttatctcac atgagaaaac acatattcgt
     5161 ttacttagtg cattcctggt cggaataacc tggccaatga gtctgcctgt ggcattactt
     5221 ttttctctct tttaggtcga cggcgccagg agacgcgtat gaacctggtg gatatcgcca
     5281 ttcttatcct caaactcatt gttgcagcac tgcaactgct tgatgctgtt ctgaaatacc
     5341 tgaagtaata cttaattaag ggcaagcgcg gggattttcc ccgcgcattt cccgggtcag
     5401 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     5461 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     5521 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     5581 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5641 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5701 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5761 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5821 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5881 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5941 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6001 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6061 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6121 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6181 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6241 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6301 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6361 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6421 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6481 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6541 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6601 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6661 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6721 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6781 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6841 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6901 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6961 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7021 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7081 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7141 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7201 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7261 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7321 atctttatag tcctgtcggg tttcgccacc tctgactcga gcgtcgattt ttgtgatgct
     7381 cgtcaggggg gcggagccta tggaaaaa
//
