LOCUS       sequence-228631-        7574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn7 tagging vector pTn7xKS with mPlum expression scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7574)
  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 7574)
  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-mPlum
FEATURES             Location/Qualifiers
     source          1..7574
                     /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(1015..1035)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             complement(1117..1797)
                     /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(1179..1202)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1377..1396)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     RBS             1805..1827
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(join(1851..1857,1858..1873,1874..1879))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    complement(1927..1974)
                     /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(2015..2020,2021..2037,2038..2043)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2232..2765
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(2895..2942)
                     /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(2988..3074)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3177..3271)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3299..3315)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3300..3316)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    3705..3726
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(3739..4821)
                     /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     4783..4802
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4960..4965,4966..4981,4982..4988)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4996..5012
                     /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             5152..5160
                     /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(5583..5602)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5702..5724
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5762..5780)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5848..5952
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5953..6021,6022..6813)
                     /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(6171..6190)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6984..7572
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 actaccggta aaccgaaagg caggaacagg agagcgcacg ggggagccgc
      781 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc
      841 gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg
      901 ccctctcact tccctgttaa gtatctccta ggccgagaaa aaaaagcccg ctcattaggc
      961 gggctagctc attatttgta gagctcatcc atgccatgtg taatcccagc agcagttaca
     1021 aactcaagaa ggaccatgtg gtcacgcttt tcgttgggat ctttcgaaag ggcagattgt
     1081 gtcgaccagc tatcgatcca tggggcccgc ggccgcctag gcgccggtgg agtggcggcc
     1141 ctcggcgcgc tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccag
     1201 cttgatgtcg gtcttgtagg cgccgggcag ctgcacgggc ttcttggcca tgtaggtggt
     1261 cttgacctcg gcgtcgtagt ggccgccgtc cttcagcctc agcctcatct tcatctcgcc
     1321 cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt
     1381 cttctgcatt acggggccgt cggaggggaa gttggtgccg cgcaccttca ccttgtagat
     1441 gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa
     1501 gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat
     1561 gtcggcgggg tgcttcacgt aggccttgga gccgtacatg atctgagggg acaggatgtc
     1621 ccaggcgaag ggcagggggc cacccttggt caccttcagc ctggcggtct gggtgccctc
     1681 gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc
     1741 catgtgctcc ttgaagcgca tgaactcctt gatgacctcc tcgcccttgc tcaccatatg
     1801 tatatctcct tcttaaagtt aaacaaaatt atttctaggg atcctccaca cattatacga
     1861 gccgatgatt aattgtcaag aatttagtga gctcatgcat gatcgaatta gcttcaaaag
     1921 cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggaa cttcaagatc
     1981 ccctgattcc ctttgtcaac agcaatggat cgaattgaca taagcctgtt cggttcgtaa
     2041 actgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag
     2101 cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttgtacagtc
     2161 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     2221 gagcagcaac gatgttacgc agcagcaacg atgttacgca gcagggcagt cgccctaaaa
     2281 caaagttagg tggctcaagt atgggcatca ttcgcacatg taggctcggc cctgaccaag
     2341 tcaaatccat gcgggctgct cttgatcttt tcggtcgtga gttcggagac gtagccacct
     2401 actcccaaca tcagccggac tccgattacc tcgggaactt gctccgtagt aagacattca
     2461 tcgcgcttgc tgccttcgac caagaagcgg ttgttggcgc tctcgcggct tacgttctgc
     2521 ccaagtttga gcagccgcgt agtgagatct atatctatga tctcgcagtc tccggcgagc
     2581 accggaggca gggcattgcc accgcgctca tcaatctcct caagcatgag gccaacgcgc
     2641 ttggtgctta tgtgatctac gtgcaagcag attacggtga cgatcccgca gtggctctct
     2701 atacaaagtt gggcatacgg gaagaagtga tgcactttga tatcgaccca agtaccgcca
     2761 cctaacaatt cgttcaagcc gagatcggct tcccggccgc ggagttgttc ggtaaattgt
     2821 cacaacgccg cggccaattc gatctagaat tccattgagt aagtttttaa gcacatcagc
     2881 ttcaaaagcg ctctgaagtt cctatacttt ctagagaata ggaacttcgg aataggtact
     2941 tcaagatccc caattcgatc gtccgggccg caagctccta gcggcggatt tgtcctactc
     3001 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc
     3061 tttcgtttta tttgatgcct caagctagag agtcattacc ccaggcgttt aagggcacca
     3121 ataactgcct taaaaaaatt acgccccgcc ctgccactca tcgcagtcta gcttggattc
     3181 tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct
     3241 gaggtcatta ctggatctat caacaggagt ccaagctcag ctaattaagc tagcttatcg
     3301 ataccgtcga cctcgaaccc cacgcccctc tttaatacga cgggcaattt gcacttcaga
     3361 aaatgaagag tttgctttag ccataacaaa agtccagtat gctttttcac agcataactg
     3421 gactgatttc agtttacaac tattctgtct agtttaagac tttattgtca tagtttagat
     3481 ctattttgtt cagtttaaga ctttattgtc cgcccacacc tgcttacgca gggcatccat
     3541 ttattactca accgtaaccg attttgccag gttacgcggc tggtcctcta gctggcgtaa
     3601 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     3661 gcgatccgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3721 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3781 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     3841 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     3901 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     3961 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctatcgagat
     4021 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     4081 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     4141 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     4201 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     4261 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     4321 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     4381 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     4441 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     4501 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     4561 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     4621 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     4681 gtaattcagc tccgccatcg ccgcttccac tttttctcgc gttttcgcag aaacgtggct
     4741 ggcctggttc accacgcgag aaacggtctg ataagagaca ccggcatact ctgcgacatc
     4801 gtatagcgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     4861 tgccatacca cgaaaggttt tgcaccattc gatgcgcctg atgcggtatt ttctccttac
     4921 gcatctgtgc ggtatttcac accgcatatc gaggagctgt tgacaattaa tcatcggctc
     4981 gtataatgtg tggaattgtg agcggataac aatttcacac ggccggccaa ggagaaaggc
     5041 tatgaagcac aaccctctgg tggtgtgtct gctcattatc tgcattacga ttctgacatt
     5101 cacactcctg acccgacaaa cgctctacga actgcggttc cgggacggtg ataaggaggt
     5161 tgctgcgctc atggcctgca cgtccaggta atcgcgagtc gacaggaagg ataagaagaa
     5221 cataaaaaat ggcactattc tctaaaatat taatttttta tgtgattggt gtgaacatat
     5281 cctttgtcat tatctggttt atctcacatg agaaaacaca tattcgttta cttagtgcat
     5341 tcctggtcgg aataacctgg ccaatgagtc tgcctgtggc attacttttt tctctctttt
     5401 aggtcgacgg cgccaggaga cgcgtatgaa cctggtggat atcgccattc ttatcctcaa
     5461 actcattgtt gcagcactgc aactgcttga tgctgttctg aaatacctga agtaatactt
     5521 aattaagggc aagcgcgggg attttccccg cgcatttccc gggtcagatg gtgcactctc
     5581 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5641 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5701 tccgggagct gcatgtgtca gaggttttca ccgtcgtcac cgaaacgcgc gagacgaaag
     5761 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5821 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5881 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5941 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6001 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6061 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6121 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6181 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6241 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6301 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6361 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6421 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6481 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6541 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6601 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6661 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6721 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6781 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6841 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6901 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6961 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7021 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7081 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     7141 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7201 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7261 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7321 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7381 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7441 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7501 gtcgggtttc gccacctctg actcgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7561 agcctatgga aaaa
//
