LOCUS       sequence-252654-        7575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-host-range GenR plasmid expressing PapRecT and EcMutL_E32K 
            with Toluic acid induction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7575)
  TITLE     Broad-host-range plasmids for improved recombineering in 
            Gammaproteobacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7575)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pORTMAGE502B
FEATURES             Location/Qualifiers
     source          1..7575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..81
                     /label=Pm promoter
                     /note="The bacterial Pm promoter is activated by XylS in 
                     the presence of benzoate or m-toluate (Marques et al., 
                     1999)."
     terminator      2906..2940
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     promoter        join(2981..2986,2987..3003,3004..3009)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3198..3731
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     oriT            3942..4050
                     /note="incP origin of transfer"
     CDS             complement(4061..4723)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGETLEPPTRVTLPRR"
     rep_origin      4724..5495
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             complement(5711..6676)
                     /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"
ORIGIN
        1 agtccagcct tgcaagaagc ggatacagga gtgcaaaaaa tggctatctc tagtaaggcc
       61 taccccttag gctttatgca acagaaacaa taataatgga gtcatgacca tgcctaggcc
      121 gcggccgcgc gaattcagaa ggagaatata ccatgggtac tgctctaacg ccgttattaa
      181 ccaagtttgc cacccgctat gagatgggaa ctacccccga agaggtcgct aatacgctga
      241 aacagacttg tttcaagggc caagtgaacg atagtcagat ggtagccctt ttgatcgttg
      301 cggatcaata taagctcaat ccattcacaa aagagctcta cgcgttccct gacaaaaata
      361 atggtattgt tccagttgtg ggagtcgatg gttgggctag aattattaac gagaatcccc
      421 agtttgatgg gatggaattc agtatggatc aacagggaac tgaatgcact tgtaaaattt
      481 accgcaaaga ccgctcgcac gccatcagcg ccaccgagta catggctgag tgcaaaagga
      541 acactcaacc ttggcagtct cacccgcgac gtatgctgcg tcataaggct atgattcaat
      601 gcgccagact agcctttggt ttcgcgggga tctacgatca ggatgaggcc gaacgcattg
      661 ttgaacgaga tgtaactccc gccgagcaat acgaggatgt atccgaagcg atttgtctga
      721 tcaaagactc accaactatg gaggacttgc aggccgcgtt ctcaaacgcg tggaaagctt
      781 acaagactaa aggtgcccgt gatcaactga ctgctgctaa agaccagaga aaaaaggagc
      841 tgttggatgc gcccattgat gtcgaattcg aagaaactgg agatgatcgt gctgcgtaag
      901 gatccaggag aatacacatg ccaattcagg tcttaccgcc acaactggcg aaccagattg
      961 ccgcaggtga ggtggtcgag cgacctgcgt cggtagtcaa agaactagtg aaaaacagcc
     1021 tcgatgcagg tgcgacgcgt atcgatattg atatcgaacg cggtggggcg aaacttatcc
     1081 gcattcgtga taacggctgc ggtatcaaaa aagatgagct ggcgctggcg ctggctcgtc
     1141 atgccaccag taaaatcgcc tctctggacg atctcgaagc cattatcagc ctgggctttc
     1201 gcggtgaggc gctggcgagt atcagttcgg tttcccgcct gacgctcact tcacgcaccg
     1261 cagaacagca ggaagcctgg caggcctatg ccgaagggcg cgatatgaac gtgacggtaa
     1321 aaccggcggc gcatcctgtg gggacgacgc tggaggtgct ggatctgttc tacaacaccc
     1381 cggcgcggcg caaattcctg cgcaccgaga aaaccgaatt taaccacatt gatgagatca
     1441 tccgccgcat tgcgctggcg cgtttcgacg tcacgatcaa cctgtcgcat aacggtaaaa
     1501 ttgtgcgtca gtaccgcgca gtgccggaag gcgggcaaaa agaacggcgc ttaggcgcga
     1561 tttgcggcac cgcttttctt gaacaagcgc tggcgattga atggcaacac ggcgatctca
     1621 cgctacgcgg ctgggtggcc gatccaaatc acaccacgcc cgcactggca gaaattcagt
     1681 attgctacgt gaacggtcgc atgatgcgcg atcgcctgat caatcacgcg atccgccagg
     1741 cctgcgaaga caaactgggg gccgatcagc aaccggcatt tgtgttgtat ctggagatcg
     1801 acccacatca ggtggacgtc aacgtgcacc ccgccaaaca cgaagtgcgt ttccatcagt
     1861 cgcgtctggt gcatgatttt atctatcagg gcgtgctgag cgtgctacaa cagcaactgg
     1921 aaacgccgct accgctggac gatgaacccc aacctgcacc gcgttccatt ccggaaaacc
     1981 gcgtggcggc ggggcgcaat cactttgcag aaccggcagc tcgtgagccg gtagctccgc
     2041 gctacactcc tgcgccagca tcaggcagtc gtccggctgc cccctggccg aatgcgcagc
     2101 caggctacca gaaacagcaa ggtgaagtgt atcgccagct tttgcaaacg cccgcgccga
     2161 tgcaaaaatt aaaagcgccg gaaccgcagg aacctgcact tgcggcgaac agtcagagtt
     2221 ttggtcgggt actgactatc gtccattccg actgtgcgtt gctggagcgc gacggcaaca
     2281 tttcactttt atccttgcca gtggcagaac gttggctgcg tcaggcacaa ttgacgccgg
     2341 gtgaagcgcc cgtttgcgcc cagccgctgc tgattccgtt gcggctaaaa gtttctgccg
     2401 aagaaaaatc ggcattagaa aaagcgcagt ctgccctggc ggaattgggt attgatttcc
     2461 agtcagatgc acagcatgtg accatcaggg cagtgccttt acccttacgc caacaaaatt
     2521 tacaaatctt gattcctgaa ctgataggct acctggcgaa gcagtccgta ttcgaacctg
     2581 gcaatattgc gcagtggatt gcacgaaatc tgatgagcga acatgcgcag tggtcaatgg
     2641 cacaggccat aaccctgctg gcggacgtgg aacggttatg tccgcaactt gtgaaaacgc
     2701 cgccgggtgg tctgttacaa tctgttgatt tacatccggc gataaaagcc ctgaaagatg
     2761 agtgatctag agtcgacctg caggcatgca agcttgcggc cgcgtcgtga ctgggaaaac
     2821 cctggcgact agtcttggac tcctgttgat agatccagta atgacctcag aactccatct
     2881 ggatttgttc agacatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat
     2941 tggtgagaat ccaggggtcc ccaataatta cgatttaaat ttgacataag cctgttcggt
     3001 tcgtaaactg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
     3061 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgt
     3121 acagcctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
     3181 gttatggagc agcaacgatg ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc
     3241 ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg
     3301 accaagtcaa atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggagacgtag
     3361 ccacctactc ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga
     3421 cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg
     3481 ttctgcccaa gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg
     3541 gagagcaccg gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca
     3601 acgcgcttgg tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg
     3661 ctctctatac aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta
     3721 ccgccaccta acaattcgtt caagccgaga tcggcttccc ggccgcggag ttgttcggta
     3781 aattggacaa cggtccgcgc gttgtccttt tccgctgcat aaccctgctt cggggtcatt
     3841 atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt ttgccaaagg
     3901 gttcgtgtag actttccttg gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta
     3961 ggcccacccg cgagcgggtg ttccttcttc actgtccctt attcgcacct ggcggtgctc
     4021 aacgggaatc ctgctctgcg aggctggccg taggccggcc ctaccggcgc ggcagcgtta
     4081 cccgtgtcgg cggctccaac gtctcgccat cgtccagaaa acacggctca tcgggcatcg
     4141 gcaggcgctg ctgcccgcgc cgttcccatt cctccgtttc ggtcaaggct ggcaggtctg
     4201 gttccatgcc cggaatgccg ggctggctgg gcggctcctc gccggggccg gtcggtagtt
     4261 gctgctcgcc cggatacagg gtcgggatgc ggcgcaggtc gccatgcccc aacagcgatt
     4321 cgtcctggtc gtcgtgatca accaccacgg cggcactgaa caccgacagg cgcaactggt
     4381 cgcggggctg gccccacgcc acgcggtcat tgaccacgta ggccgacacg gtgccggggc
     4441 cgttgagctt cacgacggag atccagcgct cggccaccaa gtccttgact gcgtattgga
     4501 ccgtccgcaa agaacgtccg atgagcttgg aaagtgtctt ctggctgacc accacggcgt
     4561 tctggtggcc catctgcgcc acgaggtgat gcagcagcat tgccgccgtg ggtttcctcg
     4621 caataagccc ggcccacgcc tcatgcgctt tgcgttccgt ttgcacccag tgaccgggct
     4681 tgttcttggc ttgaatgccg atttctctgg actgcgtggc catgcttatc tccatgcggt
     4741 aggggtgccg cacggttgcg gcaccatgcg caatcagctg caacttttcg gcagcgcgac
     4801 aacaattatg cgttgcgtaa aagtggcagt caattacaga ttttctttaa cctacgcaat
     4861 gagctattgc ggggggtgcc gcaatgagct gttgcgtacc cccctttttt aagttgttga
     4921 tttttaagtc tttcgcattt cgccctatat ctagttcttt ggtgcccaaa gaagggcacc
     4981 cctgcggggt tcccccacgc cttcggcgcg gctccccctc cggcaaaaag tggcccctcc
     5041 ggggcttgtt gatcgactgc gcggccttcg gccttgccca aggtggcgct gcccccttgg
     5101 aacccccgca ctcgccgccg tgaggctcgg ggggcaggcg ggcgggcttc gcccttcgac
     5161 tgcccccact cgcataggct tgggtcgttc caggcgcgtc aaggccaagc cgctgcgcgg
     5221 tcgctgcgcg agccttgacc cgccttccac ttggtgtcca accggcaagc gaagcgcgca
     5281 ggccgcaggc cggaggcttt tccccagaga aaattaaaaa aattgatggg gcaaggccgc
     5341 aggccgcgca gttggagccg gtgggtatgt ggtcgaaggc tgggtagccg gtgggcaatc
     5401 cctgtggtca agctcgtggg caggcgcagc ctgtccatca gcttgtccag cagggttgtc
     5461 cacgggccga gcgaagcgag ccagccggtg gccgctcgcg gccatcgtcc acatatccac
     5521 gggctggcaa gggagcgcag cgaccgcgca gggcgaagcc cggagagcaa gcccgtaggg
     5581 ggggcgcgcc cagctgtcta gggcggcgga tttgtcctac tcaggagagc gttcaccgac
     5641 aaacacgaaa ggcccagtct ttcgactgag catttcgttt tatttgatgc ctttaattaa
     5701 acgttcgtaa tcaagccact tcctttttgc attgacgcag ggtgtcggaa ggcaactcgc
     5761 cgaacgcgct cctatagttt tcagcgaagc gtcccaaatg taagaagccg tagtctaggg
     5821 ctatctcagt tatactacgc acattggcac tgggatcgtt caagcaggcg cggatgcttt
     5881 cgagcttgcg gttgcggatg tagttcttcg gcgtggtgcc ggcgtgcttc tcgaacaaat
     5941 tgtagagcga gcgtggactc atcatcgcca gctccgctaa ccgctcaagg ctgatattcc
     6001 gtttgagatt ctcctcaatg aattgaacga ctcgctcgaa agacgggtta cctttgctga
     6061 aaatttcacg gctgacattg ctgcccagca tttcgagcag cttggaagcg atgatccccg
     6121 catagtgctc ttggacccga ggcatcgact ttgtatgttc cgcttcgtca caaactaacc
     6181 cgagtagatt gataaagcca tcgagttgct ggagattgtg tcgcgcggcg aaacggatac
     6241 cctccctcgg cttgtgccaa ttgttgtcac tgcacgcccg atcaaggacc actgagggca
     6301 atttaacgat aaatttctcg caatcttctg aataggtcag gtcggcttgg tcatccggat
     6361 tgagcagcaa tagttcgccc ggcgcaaaat agtgctcctg gccatggcca cgccacaggc
     6421 aatggccttt gagtattatt tgcagatgat aacaggtctc taatccaggc gagattaccc
     6481 tcacgctacc gccgtagctg attcgacaca gatcgaggca tccgaagatt ctgtggtgca
     6541 gcctgcctgc cgggcgcccg cccttgggca ggcgaataga gtgcgtaccg acatactggt
     6601 taacataatc ggagactgca tagggctcgg cgtggacgaa gatctgactt ttctcgttca
     6661 ataagcaaaa atccatagtt cacggttctc ttattttaat gtgggctgct tggtgtgatg
     6721 tagaaaggcg ccaagtcgat gaaaatgcat ctcgacgtga tgcgtatacg ggttaccccc
     6781 attgccacgt tgcgccatcc tttttgcaat cagtgaccac ttttccaagc aaaaataacg
     6841 ccaagcagaa cgaagacgtt ctttttaaga agcgagaaca ccagaagttc gtgctgtcgg
     6901 ggcatggggc gacgaattgg cggataaagg ggatctgctg gatattacgg cctttttaaa
     6961 gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct
     7021 gatgaatgct catccgtaat tacgtatggc aatgaaagac ggtgagctgg tgatatggga
     7081 tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg
     7141 gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg
     7201 ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc
     7261 agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt
     7321 cttcgccccc gttttcacca tgcatgggaa ttagcttgat ctgaccaacg accggtagcg
     7381 gagctatcca acggcggtat accaggaaaa cacacagcag gtacatcaga acagtaccat
     7441 gactgaagaa caaatagttt tttcctgatc cataaagcag aacggcctgc tccatgacaa
     7501 atctggctcc ccaactaatg ccccatgcag ccagcataac cagcataaag tgcagtgtcc
     7561 ggtttgatag ggata
//
