LOCUS       sequence-325697-        5519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative plasmid to introduce modified rhamnose-inducible 
            promoter into bacterial genomes. Harbours enhanced rhaS binding site
            and encodes T7 gene10 stem loop in mRNA for enhanced expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5519)
  AUTHORS   Hogan AM, Jeffers KR, Palacios A, Cardona ST
  TITLE     Improved dynamic range of a rhamnose-inducible promoter for gene 
            expression in Burkholderia.
  JOURNAL   Appl Environ Microbiol. 2021 Jun 30:AEM0064721. doi: 
            10.1128/AEM.00647-21.
  PUBMED    34190606
REFERENCE   2  (bases 1 to 5519)
  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. 2021 Jun 30:AEM0064721. doi: 
            10.1128/AEM.00647-21."
COMMENT     Sequence Label: pSC201plus
FEATURES             Location/Qualifiers
     source          1..5519
                     /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"
     protein_bind    424..440
                     /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)."
     CDS             818..1654
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLDQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     CDS             1728..2576
                     /codon_start=1
                     /gene="E. coli rhaR"
                     /product="transcriptional activator of rhaR and rhaS"
                     /label=rhaR
                     /note="needed for rhamnose-dependent activation of the rhaB
                     promoter"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     promoter        join(2605..2610,2611..2627,2628..2633)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2959..3195
                     /codon_start=1
                     /gene="dhfrIIb from plasmid R388"
                     /product="E. coli plasmid-associated dihydrofolate 
                     reductase"
                     /label=TpR
                     /note="confers resistance to trimethoprim"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     primer_bind     complement(3368..3387)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3528..3550
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     oriT            4389..4498
                     /note="incP origin of transfer"
     CDS             4531..4902
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
ORIGIN
        1 gatctgaaga tcagcagttc aacctgttga tagtacgtac taagctctca tgtttcacgt
       61 actaagctct catgtttaac gtactaagct ctcatgttta acgaactaaa ccctcatggc
      121 taacgtacta agctctcatg gctaacgtac taagctctca tgtttcacgt actaagctct
      181 catgtttgaa caataaaatt aatataaatc agcaacttaa atagcctcta aggttttaag
      241 ttttataaga aaaaaaagaa tatataaggc ttttaaagct tttaaggttt aacggttgtg
      301 gacaacaagc cagggatgta acacactgag aagcccttag agcctctcaa agcaattttg
      361 agtgacacag gaacacttaa cggctgacat gggaattcca catgtggaat tccacatgtg
      421 gaattgtgag cggataacaa tttgtggaat tcccgggaga gctcgatatc gcatgcggta
      481 cctctagaag aagcttggga tccatatgtg atcctgctga atttcactag agggaaaccg
      541 ttgtggtctc ccttacgacc agtctaaaaa gcgcctgaat tcgcgacctt tccctggttg
      601 ccaggaaaat gggccattgg caaccaggga aagatgaacg tgatgatgtt cacaatttgc
      661 tgaattgtgg tgatgtgatg ctcaccgcat ttcctgaaaa ttcacgctgt atcttgaaaa
      721 atcgacgttt tttacgtggt tttccgtcga aaatttaagg taagaacctg acctcgtgat
      781 tactatttcg ccgtgttgac gacatcagga ggccagtatg accgtattac atagtgtgga
      841 tttttttccg tctggtaacg cgtccgtggc gatagaaccc cggctcccgc aggcggattt
      901 tcctgaacat catcatgatt ttcatgaaat tgtgattgtc gaacatggca cgggtattca
      961 tgtgtttaat gggcagccct ataccatcac cggtggcacg gtctgtttcg tacgcgatca
     1021 tgatcggcat ctgtatgaac ataccgataa tctgtgtctg accaatgtgc tgtatcgctc
     1081 gccggatcga tttcagtttc tcgccgggct ggatcagttg ctgccacaag agctggatgg
     1141 gcagtatccg tctcactggc gcgttaacca cagcgtattg cagcaggtgc gacagctggt
     1201 tgcacagatg gaacagcagg aaggggaaaa tgatttaccc tcgaccgcca gtcgcgagat
     1261 cttgtttatg caattactgc tcttgctgcg taaaagcagt ttgcaggaga acctggaaaa
     1321 cagcgcatca cgtctcaact tgcttctggc ctggctggag gaccattttg ccgatgaggt
     1381 gaattgggat gccgtggcgg atcaattttc tctttcactg cgtacgctac atcggcagct
     1441 taagcagcaa acgggactga cgcctcagcg atacctgaac cgcctgcgac tgatgaaagc
     1501 ccgacatctg ctacgccaca gcgaggccag cgttactgac atcgcctatc gctgtggatt
     1561 cagcgacagt aaccactttt cgacgctttt tcgccgagag tttaactggt caccgcgtga
     1621 tattcgccag ggacgggatg gctttctgca ataacgcgaa tcttctcaac gtatttgtac
     1681 gccatattgc gaataatcaa cttcgttctc tggccgaggt agccacggtg gcgcatcagt
     1741 taaaacttct caaagatgat ttttttgcca gcgaccagca ggcagtcgct gtggctgacc
     1801 gttatccgca agatgtcttt gctgaacata cacatgattt ttgtgagctg gtgattgtct
     1861 ggcgcggtaa tggcctgcat gtactcaacg atcgccctta tcgcattacc cgtggcgatc
     1921 tcttttacat tcatgctgac gataaacact cctacgcttc cgttaacgat ctggttttgc
     1981 agaatattat ttattgcccg gagcgtctga agctgaatct tgactggcag ggggcgattc
     2041 cgggatttaa cgccagcgca gggcaaccac actggcgctt aggtagcatg gggatggcgc
     2101 aggcgcggca ggttatcggt cagcttgagc atgaaagtag tcagcatgtg ccgtttgcta
     2161 acgaaatggc tgagttgctg ttcgggcagt tggtgatgtt gctgaatcgc catcgttaca
     2221 ccagtgattc gttgccgcca acatccagcg aaacgttgct ggataagctg attacccggc
     2281 tggcggctag cctgaaaagt ccctttgcgc tggataaatt ttgtgatgag gcatcgtgca
     2341 gtgagcgcgt tttgcgtcag caatttcgcc agcagactgg aatgaccatc aatcaatatc
     2401 tgcgacaggt cagagtgtgt catgcgcaat atcttctcca gcatagccgc ctgttaatca
     2461 gtgatatttc gaccgaatgt ggctttgaag atagtaacta tttttcggtg gtgtttaccc
     2521 gggaaaccgg gatgacgccc agccagtggc gtcatctcaa ttcgcagaaa gattaaatgc
     2581 atcgatgagc tcgcacgaac ccagttgaca taagcctgtt cggttcgtaa actgtaatgc
     2641 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
     2701 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttgtacagtc tatgcctcgg
     2761 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
     2821 gatgttacgc agcagggcag tcgccctaaa acaaagttag gcagccgttg tgctggtgct
     2881 ttctgatagt tgttgtgggg taggcagtca gagctcgatt tgcttgtcgc cataatagat
     2941 tcacaagaag gattcgacat gggtcaaagt agcgatgaag ccaacgctcc cgttgcaggg
     3001 cagtttgcgc ttcccctgag tgccaccttt ggcttagggg atcgcgtacg caagaaatct
     3061 ggtgccgctt ggcagggtca agtcgtcggt tggtattgca caaaactcac tcctgaaggc
     3121 tatgcggtcg agtccgaatc ccacccaggc tcagtgcaaa tttatcctgt ggctgcactt
     3181 gaacgtgtgg cctaagcgat atcttaggat ctcccatcgg tgatgtcggc gatataggcg
     3241 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     3301 tgctcatgtt tgacagctta tcatcgattt cgttccactg agcgtcagac cccgtctcga
     3361 gtctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     3421 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     3481 ggcttgcctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3541 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcaaggag atggcgccca
     3601 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     3661 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     3721 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc ctttttgtcc
     3781 ggtgttgggt tgaaggtgaa gccggtcggg gccgcagcgg gggccggctt ttcagccttg
     3841 cccccctgct tcggccgccg tggctccggc gtcttgggtg ccggcgcggg ttccgcagcc
     3901 ttggcctgcg gtgcgggcac atcggcgggc ttggccttga tgtgccgcct ggcgtgcgag
     3961 cggaacgtct cgtaggagaa cttgaccttc cccgtttccc gcatgtgctc ccaaatggtg
     4021 acgagcgcat agccggacgc taacgccgcc tcgacatccg ccctcaccgc caggaacgca
     4081 accgcagcct catcacgccg gcgcttcttg gccgcgcggg attcaaccca ctcggccagc
     4141 tcgtcggtgt agctctttgg catcgtctct cgcctgtccc ctcagttcag taatttcctg
     4201 catttgcctg tttccagtcg gtagatattc cacaaaacag cagggaagca gcgcttttcc
     4261 gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc
     4321 gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg
     4381 cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact
     4441 gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccggct
     4501 accgccggcg taacagatga gggcaagcgg atggctgatg aaaccaagcc aaccaggaag
     4561 ggcagcccac ctatcaaggt gtactgcctt ccagacgaac gaagagcgat tgaggaaaag
     4621 gcggcggcgg ccggcatgag cctgtcggcc tacctgctgg ccgtcggcca gggctacaaa
     4681 atcacgggcg tcgtggacta tgagcacgtc cgcgagctgg cccgcatcaa tggcgacctg
     4741 ggccgcctgg gcggcctgct gaaactctgg ctcaccgacg acccgcgcac ggcgcggttc
     4801 ggtgatgcca cgatcctcgc cctgctggcg aagatcgaag agaagcagga cgagcttggc
     4861 aaggtcatga tgggcgtggt ccgcccgagg gcagagccat gactttttta gccgctaaaa
     4921 cggccggggg gtgcgcgtga ttgccaagca cgtccccatg cgctccatca agaagagcga
     4981 cttcgcggag ctggtgaagt acatcaccga cgagcaaggc aagaccgagc gcctgggtca
     5041 cgtgcgcgtc acgaactgcg aggcaaacac cctgcccgct gtcatggccg aggtgatggc
     5101 gacccagcac ggcaacaccc gttccgaggc cgacaagacc tatcacctgc tggttagctt
     5161 ccgcgcggga gagaagcccg acgcggagac gttgcgcgcg attgaggacc gcatctgcgc
     5221 tgggcttggc ttcgccgagc atcagcgcgt cagtgccgtg catcacgaca ccgacaacct
     5281 gcacatccat atcgccatca acaagattca cccgacccga aacaccatcc atgagccgta
     5341 tcgggcctac cgcgccctcg ctgacctctg cgcgacgctc gaacgggact acgggcttga
     5401 gcgtgacaat cacgaaacgc ggcagcgcgt ttccgagaac cgcgcgaacg acatggagcg
     5461 gcacgcgggc gtggaaagcc tggtcggctg gatccggcca cgatgcgtcc ggcgtagag
//
