LOCUS       sequence-221657-        7577 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 7577)
  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 7577)
  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..7577
                     /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(1016..1036)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             complement(1118..1798)
                     /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(1180..1203)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1378..1397)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     RBS             1806..1828
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(join(1852..1858,1859..1874,1875..1880))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    complement(1929..1976)
                     /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(2017..2022,2023..2039,2040..2045)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2234..2767
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(2897..2944)
                     /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(2990..3076)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3179..3273)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3301..3317)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3302..3318)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    3707..3728
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(3741..4823)
                     /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     4785..4804
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4962..4967,4968..4983,4984..4990)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4998..5014
                     /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             5154..5162
                     /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(5586..5605)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5705..5727
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5765..5783)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5851..5955
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5956..6024,6025..6816)
                     /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(6174..6193)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6987..7575
                     /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 actagccggt aaaccgaaag gcaggaacag gagagcgcac gagggagccg
      781 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacca ctgatttgag
      841 cgtcagattt cgtgatgctt gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg
      901 gccctctcac ttccctgtta agtatctcct aggccgagaa aaaaaagccc gctcattagg
      961 cgggctagct cattatttgt agagctcatc catgccatgt gtaatcccag cagcagttac
     1021 aaactcaaga aggaccatgt ggtcacgctt ttcgttggga tctttcgaaa gggcagattg
     1081 tgtcgaccag ctatcgatcc atggggcccg cggccgccta ggcgccggtg gagtggcggc
     1141 cctcggcgcg ctcgtactgt tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca
     1201 gcttgatgtc ggtcttgtag gcgccgggca gctgcacggg cttcttggcc atgtaggtgg
     1261 tcttgacctc ggcgtcgtag tggccgccgt ccttcagcct cagcctcatc ttcatctcgc
     1321 ccttcagggc gccgtcctcg gggtacatcc gctcggagga ggcctcccag cccatggtct
     1381 tcttctgcat tacggggccg tcggagggga agttggtgcc gcgcaccttc accttgtaga
     1441 tgaactcgcc gtcctgcagg gaggagtcct gggtcacggt caccacgccg ccgtcctcga
     1501 agttcatcac gcgctcccac ttgaagccct cggggaagga cagcttcaag tagtcgggga
     1561 tgtcggcggg gtgcttcacg taggccttgg agccgtacat gatctgaggg gacaggatgt
     1621 cccaggcgaa gggcaggggg ccacccttgg tcaccttcag cctggcggtc tgggtgccct
     1681 cgtaggggcg gccctcgccc tcgccctcga tctcgaactc gtggccgttc acggagccct
     1741 ccatgtgctc cttgaagcgc atgaactcct tgatgacctc ctcgcccttg ctcaccatat
     1801 gtatatctcc ttcttaaagt taaacaaaat tatttctagg gatcctccac acattatacg
     1861 agccgatgat taattgtcaa gaattctagt gagctcatgc atgatcgaat tagcttcaaa
     1921 agcgctctga agttcctata ctttctagag aataggaact tcggaatagg aacttcaaga
     1981 tcccctgatt ccctttgtca acagcaatgg atcgaattga cataagcctg ttcggttcgt
     2041 aaactgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc
     2101 agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttgtacag
     2161 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
     2221 tggagcagca acgatgttac gcagcagcaa cgatgttacg cagcagggca gtcgccctaa
     2281 aacaaagtta ggtggctcaa gtatgggcat cattcgcaca tgtaggctcg gccctgacca
     2341 agtcaaatcc atgcgggctg ctcttgatct tttcggtcgt gagttcggag acgtagccac
     2401 ctactcccaa catcagccgg actccgatta cctcgggaac ttgctccgta gtaagacatt
     2461 catcgcgctt gctgccttcg accaagaagc ggttgttggc gctctcgcgg cttacgttct
     2521 gcccaggttt gagcagccgc gtagtgagat ctatatctat gatctcgcag tctccggcga
     2581 gcaccggagg cagggcattg ccaccgcgct catcaatctc ctcaagcatg aggccaacgc
     2641 gcttggtgct tatgtgatct acgtgcaagc agattacggt gacgatcccg cagtggctct
     2701 ctatacaaag ttgggcatac gggaagaagt gatgcacttt gatatcgacc caagtaccgc
     2761 cacctaacaa ttcgttcaag ccgagatcgg cttcccggcc gcggagttgt tcggtaaatt
     2821 gtcacaacgc cgcggccaat tcgatctaga attccattga gtaagttttt aagcacatca
     2881 gcttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggaa
     2941 cttcaagatc cccaattcga tcgtccgggc cgcaagctcc tagcggcgga tttgtcctac
     3001 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag
     3061 cctttcgttt tatttgatgc ctcaagctag agagtcatta ccccaggcgt ttaagggcac
     3121 caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagtc tagcttggat
     3181 tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
     3241 ctgaggtcat tactggatct atcaacagga gtccaagctc agctaattaa gctagcttat
     3301 cgataccgtc gacctcgaac cccacgcccc tctttaatac gacgggcaat ttgcacttca
     3361 gaaaatgaag agtttgcttt agccataaca aaagtccagt atgctttttc acagcataac
     3421 tggactgatt tcagtttaca actattctgt ctagtttaag actttattgt catagtttag
     3481 atctattttg ttcagtttaa gactttattg tccgcccaca cccgcttacg cagggcatcc
     3541 atttattact caaccgtaac cgattttgcc aggttacgcg gctggtcctc tagctggcgt
     3601 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     3661 tggcgatccg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3721 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3781 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     3841 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     3901 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     3961 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactatcgag
     4021 atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     4081 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     4141 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     4201 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     4261 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     4321 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     4381 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     4441 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     4501 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     4561 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     4621 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     4681 atgtaattca gctccgccat cgccgcttcc actttttctc gcgttttcgc agaaacgtgg
     4741 ctggcctggt tcaccacgcg agaaacggtc tgataagaga caccggcata ctctgcgaca
     4801 tcgtatagcg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     4861 catgccatac cacgaaaggt tttgcaccat tcgatgcgcc tgatgcggta ttttctcctt
     4921 acgcatctgt gcggtatttc acaccgcata tcgaggagct gttgacaatt aatcatcggc
     4981 tcgtataatg tgtggaattg tgagcggata acaatttcac acggccggcc aaggagaaag
     5041 gctatgaagc acaaccctct ggtggtgtgt ctgctcatta tctgcattac gattctgaca
     5101 ttcacactcc tgacccgaca aacgctctac gaactgcggt tccgggacgg tgataaggag
     5161 gttgctgcgc tcatggcctg cacgtccagg taatcgcgag tcgacaggaa ggataagaag
     5221 aacataaaaa atggcactat tctctaaaat attaattttt tatgtgattg gtgtgaacat
     5281 atcctttgtc attatctggt ttatctcaca tgagaaaaca catattcgtt tacttagtgc
     5341 attcctggtc ggaataacct ggccaatgag tctgcctgtg gcattacttt tttctctctt
     5401 ttaggtcgac ggcgccagga gacgcgtatg aacctggtgg atatcgccat tcttatcctc
     5461 aaactcattg ttgcagcact gcaactgctt gatgctgttc tgaaatacct gaagtaatac
     5521 ttaattaagg gcaagcgcgg ggattttccc cgcgcatttc ccgggtcagt atggtgcact
     5581 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     5641 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5701 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     5761 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5821 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5881 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5941 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     6001 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     6061 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6121 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     6181 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     6241 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6301 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6361 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6421 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6481 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6541 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6601 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6661 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6721 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6781 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6841 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6901 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6961 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7021 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7081 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     7141 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7201 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7261 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7321 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7381 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7441 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7501 cctgtcgggt ttcgccacct ctgactcgag cgtcgatttt tgtgatgctc gtcagggggg
     7561 cggagcctat ggaaaaa
//
