LOCUS       sequence-457262-        6620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mini-Tn7 pGP-Tn7-Cm vector with constitutive PrpsM 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 6620)
  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 6620)
  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-PrpsM
FEATURES             Location/Qualifiers
     source          1..6620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  642..807
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     promoter        1203..1307
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1308..1376,1377..2168)
                     /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(1526..1545)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(2225..2244)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2385..2407
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2546..2565
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     oriT            3246..3355
                     /note="incP origin of transfer"
     CDS             3388..3759
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      4377..4765
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     primer_bind     5160..5176
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5161..5177
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5205..5299
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      5402..5488
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    5534..5581
                     /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        5666..5768
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5769..6428
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5835..5854)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    6568..6615
                     /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)."
ORIGIN
        1 gcttttgaag ctaattcgat catgcatgag ctcgcttatc gttgttgtcg tgattatgga
       61 ctttatggct caagtgcaaa ctctgatgat gtccagtcag tatgagtctg cattgaagaa
      121 ggcgaacctg aaaggctacg gccgttaatt ggtcgcctga gaagttacgg agagtaaaaa
      181 tgaaagttcg tgcttccgtc aagaaattat gccgtaactg caaaatcgtt aagcgtgatg
      241 gtgtcatccg tgtgatttgc agtgccgagc cgaagcataa acagcgccaa ggctgatttt
      301 ttcgcatatt tttcttgcaa agttgggttg agctggctag attagccagc caatcttttg
      361 tatgtctgta cgtttccatt tgagtatcct gaaaacgggc ttttcagcat ggtacgtaca
      421 tattaaatag taggagtgca tagtggcccg tatagcaggc attaacattc ctgatcagaa
      481 acacgccgtg atcgcgttaa cttcgatcta cggtgtcggc aagacccgtt ctaaagccat
      541 cctggctgca gcgggtatcg ctgaaaatgt taagatcctc ggtacccggg cgatatctag
      601 agacgcgtac caggcatgca agcttggcgg ccgcgcaggc caaccagata agtgaaatct
      661 agttccaaac tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt
      721 cccatctatt ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc
      781 agttcccaac tattttgtcc gcccacagcg gggcattttt cttcctgtta tgtttgggcg
      841 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
      901 gcttcctcgc tcactgaccc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      961 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1021 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1081 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1141 aacccgacag gactataaac tcgacctgca gatctgcagg tggcactttt cggggaaatg
     1201 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1261 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1321 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1381 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1441 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1501 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     1561 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1621 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1681 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1741 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1801 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     1861 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1921 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1981 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2041 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     2101 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2161 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta tggtgcactc
     2221 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     2281 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     2341 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     2401 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag caaggagatg gcgcccaaca
     2461 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     2521 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     2581 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcctt tttgtccggt
     2641 gttgggttga aggtgaagcc ggtcggggcc gcagcggggg ccggcttttc agccttgccc
     2701 ccctgcttcg gccgccgtgg ctccggcgtc ttgggtgccg gcgcgggttc cgcagccttg
     2761 gcctgcggtg cgggcacatc ggcgggcttg gccttgatgt gccgcctggc gtgcgagcgg
     2821 aacgtctcgt aggagaactt gaccttcccc gtttcccgca tgtgctccca aatggtgacg
     2881 agcgcatagc cggacgctaa cgccgcctcg acatccgccc tcaccgccag gaacgcaacc
     2941 gcagcctcat cacgccggcg cttcttggcc gcgcgggatt caacccactc ggccagctcg
     3001 tcggtgtagc tctttggcat cgtctctcgc ctgtcccctc agttcagtaa tttcctgcat
     3061 ttgcctgttt ccagtcggta gatattccac aaaacagcag ggaagcagcg cttttccgct
     3121 gcataaccct gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca
     3181 cgatatacag gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt
     3241 cagccgggca ggataggtga agtaggccca cccgcgagcg ggtgttcctt cttcactgtc
     3301 ccttattcgc acctggcggt gctcaacggg aatcctgctc tgcgaggctg gccggctacc
     3361 gccggcgtaa cagatgaggg caagcggatg gctgatgaaa ccaagccaac caggaagggc
     3421 agcccaccta tcaaggtgta ctgccttcca gacgaacgaa gagcgattga ggaaaaggcg
     3481 gcggcggccg gcatgagcct gtcggcctac ctgctggccg tcggccaggg ctacaaaatc
     3541 acgggcgtcg tggactatga gcacgtccgc gagctggccc gcatcaatgg cgacctgggc
     3601 cgcctgggcg gcctgctgaa actctggctc accgacgacc cgcgcacggc gcggttcggt
     3661 gatgccacga tcctcgccct gctggcgaag atcgaagaga agcaggacga gcttggcaag
     3721 gtcatgatgg gcgtggtccg cccgagggca gagccatgac ttttttagcc gctaaaacgg
     3781 ccggggggtg cgcgtgattg ccaagcacgt ccccatgcgc tccatcaaga agagcgactt
     3841 cgcggagctg gtgaagtaca tcaccgacga gcaaggcaag accgagcgcc tgggtcacgt
     3901 gcgcgtcacg aactgcgagg caaacaccct gcccgctgtc atggccgagg tgatggcgac
     3961 ccagcacggc aacacccgtt ccgaggccga caagacctat cacctgctgg ttagcttccg
     4021 cgcgggagag aagcccgacg cggagacgtt gcgcgcgatt gaggaccgca tctgcgctgg
     4081 gcttggcttc gccgagcatc agcgcgtcag tgccgtgcat cacgacaccg acaacctgca
     4141 catccatatc gccatcaaca agattcaccc gacccgaaac accatccatg agccgtatcg
     4201 ggcctaccgc gccctcgctg acctctgcgc gacgctcgaa cgggactacg ggcttgagcg
     4261 tgacaatcac gaaacgcggc agcgcgtttc cgagaaccgc gcgaacgaca tggagcggca
     4321 cgcgggcgtg gaaagcctgg tcggctggat ccggccacga tgcgtccggc gtagaggatc
     4381 tgaagatcag cagttcaacc tgttgatagt acgtactaag ctctcatgtt tcacgtacta
     4441 agctctcatg tttaacgtac taagctctca tgtttaacga actaaaccct catggctaac
     4501 gtactaagct ctcatggcta acgtactaag ctctcatgtt tcacgtacta agctctcatg
     4561 tttgaacaat aaaattaata taaatcagca acttaaatag cctctaaggt tttaagtttt
     4621 ataagaaaaa aaagaatata taaggctttt aaagctttta aggtttaacg gttgtggaca
     4681 acaagccagg gatgtaacgc actgagaagc ccttagagcc tctcaaagca attttgagtg
     4741 acacaggaac acttaacggc tgacatggga attgtaccgc acagatgcgt aaggagaaaa
     4801 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     4861 cgggcctctt cgctattacg ccagctagag gaccagccgc gtaacctggc aaaatcggtt
     4921 acggttgagt aataaatgga tgccctgcgt aagcgggtgt gggcggacaa taaagtctta
     4981 aactgaacaa aatagatcta aactatgaca ataaagtctt aaactagaca gaatagttgt
     5041 aaactgaaat cagtccagtt atgctgtgaa aaagcatact ggacttttgt tatggctaaa
     5101 gcaaactctt cattttctga agtgcaaatt gcccgtcgta ttaaagaggg gcgtggggtt
     5161 cgaggtcgac ggtatcgata agctagctta attagctgag cttggactcc tgttgataga
     5221 tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc
     5281 gttttttatt ggtgagaatc caagctagac tgcgatgagt ggcagggcgg ggcgtaattt
     5341 ttttaaggca gttattggtg cccttaaacg cctggggtaa tgactctcta gcttgaggca
     5401 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     5461 ggtgaacgct ctcctgagta ggacaaatcc gccgctagga gcttgcggcc cggacgatcg
     5521 aattggggat cttgaagtac ctattccgaa gttcctattc tctagaaagt ataggaactt
     5581 cagagcgctt ttgaagctga tgtgcttaaa aacttactca atggaataat tctagatggg
     5641 ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac tttcaccata
     5701 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
     5761 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     5821 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     5881 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     5941 cctttattca cattcttgcc cgcctgatga atgctcatcc ggagttccgt atggcaatga
     6001 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     6061 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     6121 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     6181 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
     6241 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     6301 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     6361 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     6421 gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggttgc tacgcctgaa
     6481 taagtgataa taagcggatg aatggcagaa attcgaaagc aaattcgacc catccattgc
     6541 tgttgacaaa gggaatcagg ggatcttgaa gttcctattc cgaagttcct attctctaga
     6601 aagtatagga acttcagagc
//
