LOCUS       sequence-238455-        5867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genomic integraction of xylS-Pm->nia into the Tn7 
            insertion site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5867)
  AUTHORS   Volke DC, Turlin J, Mol V, Nikel PI
  TITLE     Physical decoupling of XylS/Pm regulatory elements and conditional 
            proteolysis enable precise control of gene expression in Pseudomonas
            putida.
  JOURNAL   Microb Biotechnol. 2019 Mar 12. doi: 10.1111/1751-7915.13383.
  PUBMED    30864281
REFERENCE   2  (bases 1 to 5867)
  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: "Microb 
            Biotechnol. 2019 Mar 12. doi: 10.1111/1751-7915.13383."
COMMENT     Sequence Label: pTn7·X-nia
FEATURES             Location/Qualifiers
     source          1..5867
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..389
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     mobile_element  398..563
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     terminator      complement(577..663)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(729..1262)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1451..1456,1457..1473,1474..1479))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             complement(1505..2470)
                     /codon_start=1
                     /gene="xylS"
                     /product="XylS regulator encoded by the Pseudomonas putida 
                     TOL plasmid pWWO"
                     /label=XylS
                     /note="stimulates transcription from the Pm promoter in the
                     presence of aromatic compounds such as m-toluate or 
                     benzoate"
                     /translation="MDFCLLNEKSQIFVHAEPYAVSDYVNQYVGTHSIRLPKGGRPAGR
                     LHHRIFGCLDLCRISYGGSVRVISPGLETCYHLQIILKGHCLWRGHGQEHYFAPGELLL
                     LNPDDQADLTYSEDCEKFIVKLPSVVLDRACSDNNWHKPREGIRFAARHNLQQLDGFIN
                     LLGLVCDEAEHTKSMPRVQEHYAGIIASKLLEMLGSNVSREIFSKGNPSFERVVQFIEE
                     NLKRNISLERLAELAMMSPRSLYNLFEKHAGTTPKNYIRNRKLESIRACLNDPSANVRS
                     ITEIALDYGFLHLGRFAENYRSAFGELPSDTLRQCKKEVA"
     promoter        3370..3450
                     /label=Pm promoter
                     /note="The bacterial Pm promoter is activated by XylS in 
                     the presence of benzoate or m-toluate (Marques et al., 
                     1999)."
     RBS             3512..3523
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      4349..4443
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4745..4764)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4756..5571
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            5738..5846
                     /note="incP origin of transfer"
ORIGIN
        1 tgtcagccgt taagtgttcc tgtgtcactc aaaattgctt tgagaggctc taagggcttc
       61 tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta aaccttaaag gctttaaaag
      121 ccttatatat tctttttttt cttataaaac ttaaaacctt agaggctatt taagttgctg
      181 atttatatta attttattgt tcaaacatga gagcttagta cgtgaaacat gagagcttag
      241 tacgttagcc atgagagctt agtacgttag ccatgagggt ttagttcgtt aaacatgaga
      301 gcttagtacg ttaaacatga gagcttagta cgtgaaacat gagagcttag tacgtactat
      361 caacaggttg aactgctgat cttcagatcg gcgcgcctgt gggcggacaa aatagttggg
      421 aactgggagg ggtggaaatg gagtttttaa ggattattta gggaagagtg acaaaataga
      481 tgggaactgg gtgtagcgtc gtaagctaat acgaaaatta aaaatgacaa aatagtttgg
      541 aactagattt cacttatctg gtttctaggg cggcggattt gtcctactca ggagagcgtt
      601 caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat
      661 ttgatgccga cgtccaattt accgaacaac tccgcggccg ggaagccgat ctcggcttga
      721 acgaattgtt aggtggcggt acttgggtcg atatcaaagt gcatcacttc ttcccgtatg
      781 cccaactttg tatagagagc cactgcggga tcgtcaccgt aatctgcttg cacgtagatc
      841 acataagcac caagcgcgtt ggcctcatgc ttgaggagat tgatgagcgc ggtggcaatg
      901 ccctgcctcc ggtgctctcc ggagactgcg agatcataga tatagatctc actacgcggc
      961 tgctcaaact tgggcagaac gtaagccgcg agagcgccaa caaccgcttc ttggtcgaag
     1021 gcagcaagcg cgatgaatgt cttactacgg agcaagttcc cgaggtaatc ggagtccggc
     1081 tgatgttggg agtaggtggc tacgtctccg aactcacgac cgaaaagatc aagagcagcc
     1141 cgcatggatt tgacttggtc agggccgagc ctacatgtgc gaatgatgcc catacttgag
     1201 ccacctaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct gctgcgtaac
     1261 atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt gctgcttgga
     1321 tgcccgaggc ataggctgta caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc
     1381 gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta
     1441 cttgcattac agtttacgaa ccgaacaggc ttatgtccaa gacgtcttaa ttaaacgttc
     1501 gtaatcaagc cacttccttt ttgcattgac gcagggtgtc ggaaggcaac tcgccgaacg
     1561 cgctcctata gttttcagcg aagcgtccca aatgtaagaa gccgtagtct agggctatct
     1621 cagttatact acgcacattg gcactgggat cgttcaagca ggcgcggatg ctttcgagct
     1681 tgcggttgcg gatgtagttc ttcggcgtgg tgccggcgtg cttctcgaac aaattgtaga
     1741 gcgagcgtgg actcatcatc gccagctccg ctaaccgctc aaggctgata ttccgtttga
     1801 gattctcctc aatgaattga acgactcgct cgaaagacgg gttacctttg ctgaaaattt
     1861 cacggctgac attgctgccc agcatttcga gcagcttgga agcgatgatc cccgcatagt
     1921 gctcttggac ccgaggcatc gactttgtat gttccgcttc gtcacaaact aacccgagta
     1981 gattgataaa gccatcgagt tgctggagat tgtgtcgcgc ggcgaaacgg ataccctccc
     2041 tcggcttgtg ccaattgttg tcactgcacg cccgatcaag gaccactgag ggcaatttaa
     2101 cgataaattt ctcgcaatct tctgaatagg tcaggtcggc ttggtcatcc ggattgagca
     2161 gcaatagttc gcccggcgca aaatagtgct cctggccatg gccacgccac aggcaatggc
     2221 ctttgagtat tatttgcaga tgataacagg tctctaatcc aggcgagatt accctcacgc
     2281 taccgccgta gctgattcga cacagatcga ggcatccgaa gattctgtgg tgcagcctgc
     2341 ctgccgggcg cccgcccttg ggcaggcgaa tagagtgcgt accgacatac tggttaacat
     2401 aatcggagac tgcatagggc tcggcgtgga cgaagatctg acttttctcg ttcaataagc
     2461 aaaaatccat agttcacggt tctcttattt taatgtgggc tgcttggtgt gatgtagaaa
     2521 ggcgccaagt cgatgaaaat gcatctcgac gtgatgcgta tacgggttac ccccattgcc
     2581 acgttgcgcc atcctttttg caatcagtga ccacttttcc aagcaaaaat aacgccaagc
     2641 agaacgaaga cgttcttttt aagaagcgag aacaccagaa gttcgtgctg tcggggcatg
     2701 gggcgacgaa ttggcggata aaggggatct gctggatatt acggcctttt taaagaccgt
     2761 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     2821 tgctcatccg taattacgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     2881 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     2941 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     3001 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     3061 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     3121 ccccgttttc accatgcatg ggaattagct tgatctgacc aacgaccggt agcggagcta
     3181 tccaacggcg gtataccagg aaaacacaca gcaggtacat cagaacagta ccatgactga
     3241 agaacaaata gttttttcct gatccataaa gcagaacggc ctgctccatg acaaatctgg
     3301 ctccccaact aatgccccat gcagccagca taaccagcat aaagtgcagt gtccggtttg
     3361 atagggataa gtccagcctt gcaagaagcg gatacaggag tgcaaaaaat ggctatctct
     3421 agtaaggcct accccttagg ctttatgcaa cagaaacaat aataatggag tcatgaccat
     3481 gcctaggccg cggccgcgcg aattcgagct caaagaggag aaattaagca tgagtaaatc
     3541 actgtttaga ggcctgagag actataatcc aatcgcaagc tcaatatgcc aattgaataa
     3601 ctcatctggt gctagacaaa gtgtaatgtt tggacttggc tttgggggtt taattgtcac
     3661 gaatcagcat ttgttcaaaa ggaatgacgg agagctaaca atccgatcgc atcatgggga
     3721 attcgtagtg aaggacacaa aaactctcaa actgcttcct tgcaaaggtc gagacatagt
     3781 gatcatcaga ttaccaaagg atttccctcc ttttccgaag aggttgcagt tccgcacccc
     3841 gacgactgag gacagagttt gtttaattgg ttcaaatttc caaacgaaga gcatttcaag
     3901 caccatgtcg gaaacaagcg caacatatcc agttgataac agtcatttct ggaaacactg
     3961 gattagcacg aaggatggtc attgcggatt acccatcgtg agcactcgag atggcagtat
     4021 tcttgggcta cacagtcttg caaattcaac gaacacccag aatttctatg cagctttccc
     4081 tgacaacttc gagaccacat acttgtcaaa tcaagacaat gataactgga taaagcagtg
     4141 gcgatacaac ccggatgaag tttgctgggg atccctacaa ctcaagaggg acattccaca
     4201 gagtccgttt acaatttgta aactgctaac ggatcttgat ggggaatttg tttacactca
     4261 gggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagcttgcgg ccgcgtcgtg
     4321 actgggaaaa ccctggcgac tagtcttgga ctcctgttga tagatccagt aatgacctca
     4381 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
     4441 aatccagcca cgcccctctt taatacgacg ggcaatttgc acttcagaaa atgaagagtt
     4501 tgctttagcc ataacaaaag tccagtatgc tttttcacag cataactgga ctgatttcag
     4561 tttacaacta ttctgtctag tttaagactt tattgtcata gtttagatct attttgttca
     4621 gtttaagact ttattgtccg cccacaattt aaatttgtgt ctcaaaatct ctgatgttac
     4681 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     4741 aatacaaggg gtgttatgag ccatattcag cgtgaaacga gctgtagccg tccgcgtctg
     4801 aacagcaaca tggatgcgga tctgtatggc tataaatggg cgcgtgataa cgtgggtcag
     4861 agcggcgcga ccatttatcg tctgtatggc aaaccggatg cgccggaact gtttctgaaa
     4921 catggcaaag gcagcgtggc gaacgatgtg accgatgaaa tggtgcgtct gaactggctg
     4981 accgaattta tgccgctgcc gaccattaaa cattttattc gcaccccgga tgatgcgtgg
     5041 ctgctgacca ccgcgattcc gggcaaaacc gcgtttcagg tgctggaaga atatccggat
     5101 agcggcgaaa acattgtgga tgcgctggcc gtgtttctgc gtcgtctgca tagcattccg
     5161 gtgtgcaact gcccgtttaa cagcgatcgt gtgtttcgtc tggcccaggc gcagagccgt
     5221 atgaacaacg gcctggtgga tgcgagcgat tttgatgatg aacgtaacgg ctggccggtg
     5281 gaacaggtgt ggaaagaaat gcataaactg ctgccgttta gcccggatag cgtggtgacc
     5341 cacggcgatt ttagcctgga taacctgatt ttcgatgaag gcaaactgat tggctgcatt
     5401 gatgtgggcc gtgtgggcat tgcggatcgt tatcaggatc tggccattct gtggaactgc
     5461 ctgggcgaat ttagcccgag cctgcaaaaa cgtctgtttc agaaatatgg cattgataat
     5521 ccggatatga acaaactgca atttcatctg atgctggatg aatttttcta ataattaatt
     5581 ggaccgcggt ccgcgcgttg tccttttccg ctgcataacc ctgcttcggg gtcattatag
     5641 cgattttttc ggtatatcca tcctttttcg cacgatatac aggattttgc caaagggttc
     5701 gtgtagactt tccttggtgt atccaacggc gtcagccggg caggataggt gaagtaggcc
     5761 cacccgcgag cgggtgttcc ttcttcactg tcccttattc gcacctggcg gtgctcaacg
     5821 ggaatcctgc tctgcgaggc tggccgtagg ccggccggcg cgcccca
//
