LOCUS       sequence-228633-        7415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn7 tagging vector pTn7xKS with sfGFP expression scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7415)
  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 7415)
  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-sfGFP
FEATURES             Location/Qualifiers
     source          1..7415
                     /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(join(922..1635,1636..1638))
                     /codon_start=1
                     /product="enhanced blue variant of GFP"
                     /label=EBFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIKV
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(967..988)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             1646..1668
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(join(1692..1698,1699..1714,1715..1720))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    complement(1768..1815)
                     /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(1856..1861,1862..1878,1879..1884)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2073..2606
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(2736..2783)
                     /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(2829..2915)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3018..3112)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3140..3156)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3141..3157)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    3546..3567
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(3580..4662)
                     /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     4624..4643
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4801..4806,4807..4822,4823..4829)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4837..4853
                     /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             4993..5001
                     /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(5424..5443)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5543..5565
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5603..5621)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5689..5793
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5794..5862,5863..6654)
                     /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(6012..6031)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6825..7413
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7314..7333
                     /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 ctcacttgta cagctcgtcc atgccgagag tgatcccggc
      961 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
     1021 gacggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     1081 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgacgt tgtggcggat
     1141 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
     1201 gttgaagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     1261 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     1321 gtaggtgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     1381 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc
     1441 gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     1501 gaacttcagg gtcagcttgc cgttggtggc atcgccctcg ccctcgccgc gcacgctgaa
     1561 cttgtggccg tttacgtcgc catccagctc gaccaggatg ggcaccaccc cggtgaacag
     1621 ctcctcgccc ttgctcatat gtatatctcc ttcttaaagt taaacaaaat tatttctagg
     1681 gatcctccac acattatacg agccgatgat taattgtcaa gaatttagtg agctcatgca
     1741 tgatcgaatt agcttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt
     1801 cggaatagga acttcaagat cccctgattc cctttgtcaa cagcaatgga tcgaattgac
     1861 ataagcctgt tcggttcgta aactgtaatg caagtagcgt atgcgctcac gcaactggtc
     1921 cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta
     1981 tgactgtttt tttgtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt
     2041 gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagcaac gatgttacgc
     2101 agcagggcag tcgccctaaa acaaagttag gtggctcaag tatgggcatc attcgcacat
     2161 gtaggctcgg ccctgaccaa gtcaaatcca tgcgggctgc tcttgatctt ttcggtcgtg
     2221 agttcggaga cgtagccacc tactcccaac atcagccgga ctccgattac ctcgggaact
     2281 tgctccgtag taagacattc atcgcgcttg ctgccttcga ccaagaagcg gttgttggcg
     2341 ctctcgcggc ttacgttctg cccaagtttg agcagccgcg tagtgagatc tatatctatg
     2401 atctcgcagt ctccggcgag caccggaggc agggcattgc caccgcgctc atcaatctcc
     2461 tcaagcatga ggccaacgcg cttggtgctt atgtgatcta cgtgcaagca gattacggtg
     2521 acgatcccgc agtggctctc tatacaaagt tgggcatacg ggaagaagtg atgcactttg
     2581 atatcgaccc aagtaccgcc acctaacaat tcgttcaagc cgagatcggc ttcccggccg
     2641 cggagttgtt cggtaaattg tcacaacgcc gcggccaatt cgatctagaa ttccattgag
     2701 taagttttta agcacatcag cttcaaaagc gctctgaagt tcctatactt tctagagaat
     2761 aggaacttcg gaataggtac ttcaagatcc ccaattcgat cgtccgggcc gcaagctcct
     2821 agcggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
     2881 gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tcaagctaga gagtcattac
     2941 cccaggcgtt taagggcacc aataactgcc ttaaaaaaat tacgccccgc cctgccactc
     3001 atcgcagtct agcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg
     3061 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagctca
     3121 gctaattaag ctagcttatc gataccgtcg acctcgaacc ccacgcccct ctttaatacg
     3181 acgggcaatt tgcacttcag aaaatgaaga gtttgcttta gccataacaa aagtccagta
     3241 tgctttttca cagcataact ggactgattt cagtttacaa ctattctgtc tagtttaaga
     3301 ctttattgtc atagtttaga tctattttgt tcagtttaag actttattgt ccgcccacac
     3361 ctgcttacgc agggcatcca tttattactc aaccgtaacc gattttgcca ggttacgcgg
     3421 ctggtcctct agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     3481 gcgcagcctg aatggcgaat ggcgatccga gccggaagca taaagtgtaa agcctggggt
     3541 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3601 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3661 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     3721 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     3781 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     3841 gtcgtatccc actatcgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
     3901 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     3961 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     4021 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     4081 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     4141 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     4201 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     4261 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     4321 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     4381 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     4441 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     4501 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttctcg
     4561 cgttttcgca gaaacgtggc tggcctggtt caccacgcga gaaacggtct gataagagac
     4621 accggcatac tctgcgacat cgtatagcgt tactggtttc acattcacca ccctgaattg
     4681 actctcttcc gggcgctatc atgccatacc acgaaaggtt ttgcaccatt cgatgcgcct
     4741 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat cgaggagctg
     4801 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
     4861 cggccggcca aggagaaagg ctatgaagca caaccctctg gtggtgtgtc tgctcattat
     4921 ctgcattacg attctgacat tcacactcct gacccgacaa acgctctacg aactgcggtt
     4981 ccgggacggt gataaggagg ttgctgcgct catggcctgc acgtccaggt aatcgcgagt
     5041 cgacaggaag gataagaaga acataaaaaa tggcactatt ctctaaaata ttaatttttt
     5101 atgtgattgg tgtgaacata tcctttgtca ttatctggtt tatctcacat gagaaaacac
     5161 atattcgttt acttagtgca ttcctggtcg gaataacctg gccaatgagt ctgcctgtgg
     5221 cattactttt ttctctcttt taggtcgacg gcgccaggag acgcgtatga acctggtgga
     5281 tatcgccatt cttatcctca aactcattgt tgcagcactg caactgcttg atgctgttct
     5341 gaaatacctg aagtaatact taattaaggg caagcgcggg gattttcccc gcgcatttcc
     5401 cgggtcagat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     5461 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5521 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcgtca
     5581 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5641 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5701 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5761 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5821 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5881 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5941 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     6001 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     6061 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6121 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6181 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6241 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6301 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     6361 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6421 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6481 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6541 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6601 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6661 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6721 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6781 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6841 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6901 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6961 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7021 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7081 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7141 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7201 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7261 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7321 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gactcgagcg tcgatttttg
     7381 tgatgctcgt caggggggcg gagcctatgg aaaaa
//
