LOCUS       sequence-457263-        6201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mini-Tn7 pGP-Tn7-Cm vector with constitutive PcL promoter for the 
            constitutive expression of target genes inserted into the attTn7 
            chromosomal site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6201)
  AUTHORS   Chang C, Phan M-D, Schembri MA
  TITLE     Modified Tn7 transposon vectors for controlled chromosomal gene 
            expression.
  JOURNAL   Appl Environ Microbiol. 2024 Oct 23;90(10):e0155624. doi: 
            10.1128/aem.01556-24. Epub 2024 Sep 18.
  PUBMED    39291982
REFERENCE   2  (bases 1 to 6201)
  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: "Appl 
            Environ Microbiol."; date: "2024-10-23"; volume: "90(10):e0155624. 
            doi"; pages: " 10.1128/aem.01556-24. Epub 2024 Sep 18"
COMMENT     Sequence Label: pGP-Tn7-PcL
FEATURES             Location/Qualifiers
     source          1..6201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    158..205
                     /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)."
     primer_bind     259..277
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     mobile_element  433..598
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     promoter        994..1098
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1099..1167,1168..1959)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1317..1336)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(2016..2035)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2176..2198
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2337..2356
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     oriT            3037..3146
                     /note="incP origin of transfer"
     CDS             3179..3550
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      4168..4556
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     primer_bind     4951..4967
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4952..4968
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4996..5090
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      5193..5279
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    5325..5372
                     /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        5457..5559
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(5560..6201,1..18)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5626..5645)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggttgc
       61 tacgcctgaa taagtgataa taagcggatg aatggcagaa attcgaaagc aaattcgacc
      121 catccattgc tgttgacaaa gggaatcagg ggatcttgaa gttcctattc cgaagttcct
      181 attctctaga aagtatagga acttcagagc gcttttgaag ctaattcgat catgcatgag
      241 ctcactagtg gatcccccaa taggcgtatc acgaggccct ttcgtcttca cctcgaggat
      301 aaatatctaa caccgtgcgt gttgactatt ttacctctgg cggtgataat ggttgcatgt
      361 actagaattc cggtacccgg gcgatatcta gagacgcgta ccaggcatgc aagcttggcg
      421 gccgcgcagg ccaaccagat aagtgaaatc tagttccaaa ctattttgtc atttttaatt
      481 ttcgtattag cttacgacgc tacacccagt tcccatctat tttgtcactc ttccctaaat
      541 aatccttaaa aactccattt ccacccctcc cagttcccaa ctattttgtc cgcccacagc
      601 ggggcatttt tcttcctgtt atgtttgggc gctgcattaa tgaatcggcc aacgcgcggg
      661 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgacc cgctgcgctc
      721 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
      781 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
      841 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      901 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa ctcgacctgc
      961 agatctgcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1021 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1081 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1141 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1201 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1261 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1321 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     1381 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1441 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1501 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1561 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     1621 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     1681 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     1741 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     1801 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     1861 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     1921 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     1981 tatatacttt agattgattt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     2041 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     2101 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2161 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     2221 gcgcgaggca gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     2281 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     2341 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     2401 cgtccggcgt agaggatcct ttttgtccgg tgttgggttg aaggtgaagc cggtcggggc
     2461 cgcagcgggg gccggctttt cagccttgcc cccctgcttc ggccgccgtg gctccggcgt
     2521 cttgggtgcc ggcgcgggtt ccgcagcctt ggcctgcggt gcgggcacat cggcgggctt
     2581 ggccttgatg tgccgcctgg cgtgcgagcg gaacgtctcg taggagaact tgaccttccc
     2641 cgtttcccgc atgtgctccc aaatggtgac gagcgcatag ccggacgcta acgccgcctc
     2701 gacatccgcc ctcaccgcca ggaacgcaac cgcagcctca tcacgccggc gcttcttggc
     2761 cgcgcgggat tcaacccact cggccagctc gtcggtgtag ctctttggca tcgtctctcg
     2821 cctgtcccct cagttcagta atttcctgca tttgcctgtt tccagtcggt agatattcca
     2881 caaaacagca gggaagcagc gcttttccgc tgcataaccc tgcttcgggg tcattatagc
     2941 gattttttcg gtatatccat cctttttcgc acgatataca ggattttgcc aaagggttcg
     3001 tgtagacttt ccttggtgta tccaacggcg tcagccgggc aggataggtg aagtaggccc
     3061 acccgcgagc gggtgttcct tcttcactgt cccttattcg cacctggcgg tgctcaacgg
     3121 gaatcctgct ctgcgaggct ggccggctac cgccggcgta acagatgagg gcaagcggat
     3181 ggctgatgaa accaagccaa ccaggaaggg cagcccacct atcaaggtgt actgccttcc
     3241 agacgaacga agagcgattg aggaaaaggc ggcggcggcc ggcatgagcc tgtcggccta
     3301 cctgctggcc gtcggccagg gctacaaaat cacgggcgtc gtggactatg agcacgtccg
     3361 cgagctggcc cgcatcaatg gcgacctggg ccgcctgggc ggcctgctga aactctggct
     3421 caccgacgac ccgcgcacgg cgcggttcgg tgatgccacg atcctcgccc tgctggcgaa
     3481 gatcgaagag aagcaggacg agcttggcaa ggtcatgatg ggcgtggtcc gcccgagggc
     3541 agagccatga cttttttagc cgctaaaacg gccggggggt gcgcgtgatt gccaagcacg
     3601 tccccatgcg ctccatcaag aagagcgact tcgcggagct ggtgaagtac atcaccgacg
     3661 agcaaggcaa gaccgagcgc ctgggtcacg tgcgcgtcac gaactgcgag gcaaacaccc
     3721 tgcccgctgt catggccgag gtgatggcga cccagcacgg caacacccgt tccgaggccg
     3781 acaagaccta tcacctgctg gttagcttcc gcgcgggaga gaagcccgac gcggagacgt
     3841 tgcgcgcgat tgaggaccgc atctgcgctg ggcttggctt cgccgagcat cagcgcgtca
     3901 gtgccgtgca tcacgacacc gacaacctgc acatccatat cgccatcaac aagattcacc
     3961 cgacccgaaa caccatccat gagccgtatc gggcctaccg cgccctcgct gacctctgcg
     4021 cgacgctcga acgggactac gggcttgagc gtgacaatca cgaaacgcgg cagcgcgttt
     4081 ccgagaaccg cgcgaacgac atggagcggc acgcgggcgt ggaaagcctg gtcggctgga
     4141 tccggccacg atgcgtccgg cgtagaggat ctgaagatca gcagttcaac ctgttgatag
     4201 tacgtactaa gctctcatgt ttcacgtact aagctctcat gtttaacgta ctaagctctc
     4261 atgtttaacg aactaaaccc tcatggctaa cgtactaagc tctcatggct aacgtactaa
     4321 gctctcatgt ttcacgtact aagctctcat gtttgaacaa taaaattaat ataaatcagc
     4381 aacttaaata gcctctaagg ttttaagttt tataagaaaa aaaagaatat ataaggcttt
     4441 taaagctttt aaggtttaac ggttgtggac aacaagccag ggatgtaacg cactgagaag
     4501 cccttagagc ctctcaaagc aattttgagt gacacaggaa cacttaacgg ctgacatggg
     4561 aattgtaccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     4621 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctaga
     4681 ggaccagccg cgtaacctgg caaaatcggt tacggttgag taataaatgg atgccctgcg
     4741 taagcgggtg tgggcggaca ataaagtctt aaactgaaca aaatagatct aaactatgac
     4801 aataaagtct taaactagac agaatagttg taaactgaaa tcagtccagt tatgctgtga
     4861 aaaagcatac tggacttttg ttatggctaa agcaaactct tcattttctg aagtgcaaat
     4921 tgcccgtcgt attaaagagg ggcgtggggt tcgaggtcga cggtatcgat aagctagctt
     4981 aattagctga gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg
     5041 atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagctaga
     5101 ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac
     5161 gcctggggta atgactctct agcttgaggc atcaaataaa acgaaaggct cagtcgaaag
     5221 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     5281 cgccgctagg agcttgcggc ccggacgatc gaattgggga tcttgaagta cctattccga
     5341 agttcctatt ctctagaaag tataggaact tcagagcgct tttgaagctg atgtgcttaa
     5401 aaacttactc aatggaataa ttctagatgg gccaactttt ggcgaaaatg agacgttgat
     5461 cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt
     5521 tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     5581 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     5641 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     5701 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     5761 aatgctcatc cggagttccg tatggcaatg aaagacggtg agctggtgat atgggatagt
     5821 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     5881 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     5941 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     6001 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     6061 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     6121 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     6181 ttacaacagt actgcgatga g
//
