LOCUS       sequence-248572-        5461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A variant of the pSCrhaB2 vector with the oriR6K and mob genes from 
            pGP-Omega-TP. Used for homologous recombination of the 
            rhamnose-inducible system into the chromosome of Burkholderia 
            cenocepacia K56-2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5461)
  AUTHORS   Ortega XP, Cardona ST, Brown AR, Loutet SA, Flannagan RS, Campopiano
            DJ, Govan JR, Valvano MA
  TITLE     A putative gene cluster for aminoarabinose biosynthesis is essential
            for Burkholderia cenocepacia viability.
  JOURNAL   J Bacteriol. 2007 May;189(9):3639-44. doi: 10.1128/JB.00153-07. Epub
            2007 Mar 2.
  PUBMED    17337576
REFERENCE   2  (bases 1 to 5461)
  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"; doi: 
            "10.1128/JB.00153-07"; journalName: "J Bacteriol"; date: "2007-05"; 
            volume: "189"; issue: "9"; pages: "3639-44"
COMMENT     Sequence Label: pSC201
FEATURES             Location/Qualifiers
     source          1..5461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        126..244
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     rep_origin      384..740
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            1384..1493
                     /note="incP origin of transfer"
     CDS             1526..1897
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     primer_bind     complement(2706..2728)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2869..2888
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(3061..3297)
                     /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"
     promoter        complement(join(3624..3629,3630..3646,3647..3652))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             complement(3683..4531)
                     /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"
     CDS             complement(4605..5441)
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
ORIGIN
        1 aacacggcga aatagtaatc acgaggtcag gttcttacct taaattttcg acggaaaacc
       61 acgtaaaaaa cgtcgatttt tcaagataca gcgtgaattt tcaggaaatg cggtgagcat
      121 cacatcacca caattcagca aattgtgaac atcatcacgt tcatctttcc ctggttgcca
      181 atggcccatt ttcctgtcag taacgagaag gtcgcgaatt caggcgcttt ttagactggt
      241 cgtaatgaaa ttcagcagga tcacatatgg atcccaagct tcttctagag gtaccgcatg
      301 cgatatcgag ctctcccggg aattcgatcg ctagtttgtt ttgactccat ccattagggc
      361 ttctaaaacg ccttctaagg ccatgtcagc cgttaagtgt tcctgtgtca ctgaaaattg
      421 ctttgagagg ctctaagggc ttctcagtgc gttacatccc tggcttgttg tccacaaccg
      481 ttaaacctta aaagctttaa aagccttata tattcttttt tttcttataa aacttaaaac
      541 cttagaggct atttaagttg ctgatttata ttaattttat tgttcaaaca tgagagctta
      601 gtacgtgaaa catgagagct tagtacgtta gccatgagag cttagtacgt tagccatgag
      661 ggtttagttc gttaaacatg agagcttagt acgttaaaca tgagagctta gtacgtgaaa
      721 catgagagct tagtacgtac tatcaacagg ttgaactgct ggatcctttt tgtccggtgt
      781 tgggttgaag gtgaagccgg tcggggccgc agcgggggcc ggcttttcag ccttgccccc
      841 ctgcttcggc cgccgtggct ccggcgtctt gggtgccggc gcgggttccg cagccttggc
      901 ctgcggtgcg ggcacatcgg cgggcttggc cttgatgtgc cgcctggcgt gcgagcggaa
      961 cgtctcgtag gagaacttga ccttccccgt ttcccgcatg tgctcccaaa tggtgacgag
     1021 cgcatagccg gacgctaacg ccgcctcgac atccgccctc accgccagga acgcaaccgc
     1081 agcctcatca cgccggcgct tcttggccgc gcgggattca acccactcgg ccagctcgtc
     1141 ggtgtagctc tttggcatcg tctctcgcct gtcccctcag ttcagtaatt tcctgcattt
     1201 gcctgtttcc agtcggtaga tattccacaa aacagcaggg aagcagcgct tttccgctgc
     1261 ataaccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg
     1321 atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca
     1381 gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc
     1441 ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cggctaccgc
     1501 cggcgtaaca gatgagggca agcggatggc tgatgaaacc aagccaacca ggaagggcag
     1561 cccacctatc aaggtgtact gccttccaga cgaacgaaga gcgattgagg aaaaggcggc
     1621 ggcggccggc atgagcctgt cggcctacct gctggccgtc ggccagggct acaaaatcac
     1681 gggcgtcgtg gactatgagc acgtccgcga gctggcccgc atcaatggcg acctgggccg
     1741 cctgggcggc ctgctgaaac tctggctcac cgacgacccg cgcacggcgc ggttcggtga
     1801 tgccacgatc ctcgccctgc tggcgaagat cgaagagaag caggacgagc ttggcaaggt
     1861 catgatgggc gtggtccgcc cgagggcaga gccatgactt ttttagccgc taaaacggcc
     1921 ggggggtgcg cgtgattgcc aagcacgtcc ccatgcgctc catcaagaag agcgacttcg
     1981 cggagctggt gaagtacatc accgacgagc aaggcaagac cgagcgcctg ggtcacgtgc
     2041 gcgtcacgaa ctgcgaggca aacaccctgc ccgctgtcat ggccgaggtg atggcgaccc
     2101 agcacggcaa cacccgttcc gaggccgaca agacctatca cctgctggtt agcttccgcg
     2161 cgggagagaa gcccgacgcg gagacgttgc gcgcgattga ggaccgcatc tgcgctgggc
     2221 ttggcttcgc cgagcatcag cgcgtcagtg ccgtgcatca cgacaccgac aacctgcaca
     2281 tccatatcgc catcaacaag attcacccga cccgaaacac catccatgag ccgtatcggg
     2341 cctaccgcgc cctcgctgac ctctgcgcga cgctcgaacg ggactacggg cttgagcgtg
     2401 acaatcacga aacgcggcag cgcgtttccg agaaccgcgc gaacgacatg gagcggcacg
     2461 cgggcgtgga aagcctggtc ggctggatcc tctacgccgg acgcatcgtg gccggcatca
     2521 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
     2581 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg
     2641 tggccggggg actgttgggc gccatctcct tgctgcctcg cgcgtttcgg tgatgacggt
     2701 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     2761 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
     2821 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
     2881 agattgtact gagactcgag acggggtctg acgctcagtg gaacgaaatc gatgataagc
     2941 tgtcaaacat gagcagatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
     3001 aggtgcggtt gctggcgcct atatcgccga catcaccgat gggagatcct aagatatcgc
     3061 ttaggccaca cgttcaagtg cagccacagg ataaatttgc actgagcctg ggtgggattc
     3121 ggactcgacc gcatagcctt caggagtgag ttttgtgcaa taccaaccga cgacttgacc
     3181 ctgccaagcg gcaccagatt tcttgcgtac gcgatcccct aagccaaagg tggcactcag
     3241 gggaagcgca aactgccctg caacgggagc gttggcttca tcgctacttt gacccatgtc
     3301 gaatccttct tgtgaatcta ttatggcgac aagcaaatcg agctctgact gcctacccca
     3361 caacaactac tagaaagcac cagcacaacg gctgcctaac tttgttttag ggcgactacc
     3421 ctgctgcgta acatcgttgc tgttccataa catcaaacat cgacccacgg cgtaacgcgc
     3481 ttagctagct tggatgcccg aggctagact gtacaaaaaa acagtcataa caagccatga
     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
     3661 gtgcgagctc atcgatatgc atttaatctt tctgcgaatt gagatgacgc cactggctgg
     3721 gcgtcatccc ggtttcccgg gtaaacacca ccgaaaaata gttactatct tcaaagccac
     3781 attcggtcga aatatcactg attaacaggc ggctatgctg gagaagatat tgcgcatgac
     3841 acactctgac ctgtcgcaga tattgattga tggtcattcc agtctgctgg cgaaattgct
     3901 gacgcaaaac gcgctcactg cacgatgcct catcacaaaa tttatccagc gcaaagggac
     3961 ttttcaggct agccgccagc cgggtaatca gcttatccag caacgtttcg ctggatgttg
     4021 gcggcaacga atcactggtg taacgatggc gattcagcaa catcaccaac tgcccgaaca
     4081 gcaactcagc catttcgtta gcaaacggca catgctgact actttcatgc tcaagctgac
     4141 cgataacctg ccgcgcctgc gccatcccca tgctacctaa gcgccagtgt ggttgccctg
     4201 cgctggcgtt aaatcccgga atcgccccct gccagtcaag attcagcttc agacgctccg
     4261 ggcaataaat aatattctgc aaaaccagat cgttaacgga agcgtaggag tgtttatcgt
     4321 cagcatgaat gtaaaagaga tcgccacggg taatgcgata agggcgatcg ttgagtacat
     4381 gcaggccatt accgcgccag acaatcacca gctcacaaaa atcatgtgta tgttcagcaa
     4441 agacatcttg cggataacgg tcagccacag cgactgcctg ctggtcgctg gcaaaaaaat
     4501 catctttgag aagttttaac tgatgcgcca ccgtggctac ctcggccaga gaacgaagtt
     4561 gattattcgc aatatggcgt acaaatacgt tgagaagatt cgcgttattg cagaaagcca
     4621 tcccgtccct ggcgaatatc acgcggtgac cagttaaact ctcggcgaaa aagcgtcgaa
     4681 aagtggttac tgtcgctgaa tccacagcga taggcgatgt cagtaacgct ggcctcgctg
     4741 tggcgtagca gatgtcgggc tttcatcagt cgcaggcggt tcaggtatcg ctgaggcgtc
     4801 agtcccgttt gctgcttaag ctgccgatgt agcgtacgca gtgaaagaga aaattgatcc
     4861 gccacggcat cccaattcac ctcatcggca aaatggtcct ccagccaggc cagaagcaag
     4921 ttgagacgtg atgcgctgtt ttccaggttc tcctgcaaac tgcttttacg cagcaagagc
     4981 agtaattgca taaacaagat ctcgcgactg gcggtcgagg gtaaatcatt ttccccttcc
     5041 tgctgttcca tctgtgcaac cagctgtcgc acctgctgca atacgctgtg gttaacgcgc
     5101 cagtgagacg gatactgccc atccagctct tgtggcagca actgattcag cccggcgaga
     5161 aactgaaatc gatccggcga gcgatacagc acattggtca gacacagatt atcggtatgt
     5221 tcatacagat gccgatcatg atcgcgtacg aaacagaccg tgccaccggt gatggtatag
     5281 ggctgcccat taaacacatg aatacccgtg ccatgttcga caatcacaat ttcatgaaaa
     5341 tcatgatgat gttcaggaaa atccgcctgc gggagccggg gttctatcgc cacggacgcg
     5401 ttaccagacg gaaaaaaatc cacactatgt aatacggtca tactggcctc ctgatgtcgt
     5461 c
//
