LOCUS       sequence-221653-        7420 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 7420)
  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 7420)
  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..7420
                     /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(join(925..1638,1639..1641))
                     /codon_start=1
                     /product="enhanced blue variant of GFP"
                     /label=EBFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIKV
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(970..991)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             1649..1671
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(join(1695..1701,1702..1717,1718..1723))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    complement(1772..1819)
                     /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(1860..1865,1866..1882,1883..1888)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2077..2610
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(2740..2787)
                     /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(2833..2919)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3022..3116)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3144..3160)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3145..3161)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    3550..3571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(3584..4666)
                     /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     4628..4647
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4805..4810,4811..4826,4827..4833)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4841..4857
                     /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             4997..5005
                     /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(5429..5448)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5548..5570
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5608..5626)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5694..5798
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5799..5867,5868..6659)
                     /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(6017..6036)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6830..7418
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7319..7338
                     /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 ccgctcactt gtacagctcg tccatgccga gagtgatccc
      961 ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct
     1021 caggacggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat
     1081 gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga cgttgtggcg
     1141 gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg
     1201 gctgttgaag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
     1261 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
     1321 cttgtaggtg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat
     1381 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac
     1441 gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
     1501 gatgaacttc agggtcagct tgccgttggt ggcatcgccc tcgccctcgc cgcgcacgct
     1561 gaacttgtgg ccgtttacgt cgccatccag ctcgaccagg atgggcacca ccccggtgaa
     1621 cagctcctcg cccttgctca tatgtatatc tccttcttaa agttaaacaa aattatttct
     1681 agggatcctc cacacattat acgagccgat gattaattgt caagaattct agtgagctca
     1741 tgcatgatcg aattagcttc aaaagcgctc tgaagttcct atactttcta gagaatagga
     1801 acttcggaat aggaacttca agatcccctg attccctttg tcaacagcaa tggatcgaat
     1861 tgacataagc ctgttcggtt cgtaaactgt aatgcaagta gcgtatgcgc tcacgcaact
     1921 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
     1981 gttatgactg tttttttgta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg
     2041 ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag caacgatgtt
     2101 acgcagcagg gcagtcgccc taaaacaaag ttaggtggct caagtatggg catcattcgc
     2161 acatgtaggc tcggccctga ccaagtcaaa tccatgcggg ctgctcttga tcttttcggt
     2221 cgtgagttcg gagacgtagc cacctactcc caacatcagc cggactccga ttacctcggg
     2281 aacttgctcc gtagtaagac attcatcgcg cttgctgcct tcgaccaaga agcggttgtt
     2341 ggcgctctcg cggcttacgt tctgcccagg tttgagcagc cgcgtagtga gatctatatc
     2401 tatgatctcg cagtctccgg cgagcaccgg aggcagggca ttgccaccgc gctcatcaat
     2461 ctcctcaagc atgaggccaa cgcgcttggt gcttatgtga tctacgtgca agcagattac
     2521 ggtgacgatc ccgcagtggc tctctataca aagttgggca tacgggaaga agtgatgcac
     2581 tttgatatcg acccaagtac cgccacctaa caattcgttc aagccgagat cggcttcccg
     2641 gccgcggagt tgttcggtaa attgtcacaa cgccgcggcc aattcgatct agaattccat
     2701 tgagtaagtt tttaagcaca tcagcttcaa aagcgctctg aagttcctat actttctaga
     2761 gaataggaac ttcggaatag gaacttcaag atccccaatt cgatcgtccg ggccgcaagc
     2821 tcctagcggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     2881 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctcaagc tagagagtca
     2941 ttaccccagg cgtttaaggg caccaataac tgccttaaaa aaattacgcc ccgccctgcc
     3001 actcatcgca gtctagcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt
     3061 tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag
     3121 ctcagctaat taagctagct tatcgatacc gtcgacctcg aaccccacgc ccctctttaa
     3181 tacgacgggc aatttgcact tcagaaaatg aagagtttgc tttagccata acaaaagtcc
     3241 agtatgcttt ttcacagcat aactggactg atttcagttt acaactattc tgtctagttt
     3301 aagactttat tgtcatagtt tagatctatt ttgttcagtt taagacttta ttgtccgccc
     3361 acacccgctt acgcagggca tccatttatt actcaaccgt aaccgatttt gccaggttac
     3421 gcggctggtc ctctagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     3481 agttgcgcag cctgaatggc gaatggcgat ccgagccgga agcataaagt gtaaagcctg
     3541 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3601 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3661 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     3721 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     3781 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     3841 tatcgtcgta tcccactatc gagatatccg caccaacgcg cagcccggac tcggtaatgg
     3901 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     3961 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     4021 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     4081 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     4141 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     4201 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     4261 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     4321 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     4381 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     4441 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     4501 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     4561 ctcgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgagaaacg gtctgataag
     4621 agacaccggc atactctgcg acatcgtata gcgttactgg tttcacattc accaccctga
     4681 attgactctc ttccgggcgc tatcatgcca taccacgaaa ggttttgcac cattcgatgc
     4741 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatcgagga
     4801 gctgttgaca attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt
     4861 cacacggccg gccaaggaga aaggctatga agcacaaccc tctggtggtg tgtctgctca
     4921 ttatctgcat tacgattctg acattcacac tcctgacccg acaaacgctc tacgaactgc
     4981 ggttccggga cggtgataag gaggttgctg cgctcatggc ctgcacgtcc aggtaatcgc
     5041 gagtcgacag gaaggataag aagaacataa aaaatggcac tattctctaa aatattaatt
     5101 ttttatgtga ttggtgtgaa catatccttt gtcattatct ggtttatctc acatgagaaa
     5161 acacatattc gtttacttag tgcattcctg gtcggaataa cctggccaat gagtctgcct
     5221 gtggcattac ttttttctct cttttaggtc gacggcgcca ggagacgcgt atgaacctgg
     5281 tggatatcgc cattcttatc ctcaaactca ttgttgcagc actgcaactg cttgatgctg
     5341 ttctgaaata cctgaagtaa tacttaatta agggcaagcg cggggatttt ccccgcgcat
     5401 ttcccgggtc agtatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5461 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5521 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5581 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5641 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5701 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5761 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5821 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5881 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5941 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6001 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6061 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6121 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6181 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6241 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6301 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6361 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6421 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6481 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6541 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6601 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6661 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6721 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6781 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6841 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6901 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6961 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7021 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7081 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7141 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7201 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7261 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7321 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactc gagcgtcgat
     7381 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
//
