LOCUS       sequence-257366-        5493 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 5493)
  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 5493)
  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..5493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        539..657
                     /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)"
     protein_bind    745..761
                     /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)."
     rep_origin      796..1184
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(1802..2173)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            2206..2315
                     /note="incP origin of transfer"
     primer_bind     complement(3154..3176)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3317..3336
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(3509..3745)
                     /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(4071..4076,4077..4093,4094..4099))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             complement(4128..4976)
                     /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(join(5050..5493,1..393))
                     /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"
ORIGIN
        1 catctgtgca accagctgtc gcacctgctg caatacgctg tggttaacgc gccagtgaga
       61 cggatactgc ccatccagct cttgtggcag caactgatcc agcccggcga gaaactgaaa
      121 tcgatccggc gagcgataca gcacattggt cagacacaga ttatcggtat gttcatacag
      181 atgccgatca tgatcgcgta cgaaacagac cgtgccaccg gtgatggtat agggctgccc
      241 attaaacaca tgaatacccg tgccatgttc gacaatcaca atttcatgaa aatcatgatg
      301 atgttcagga aaatccgcct gcgggagccg gggttctatc gccacggacg cgttaccaga
      361 cggaaaaaaa tccacactat gtaatacggt catactggcc tcctgatgtc gtcaacacgg
      421 cgaaatagta atcacgaggt caggttctta ccttaaattt tcgacggaaa accacgtaaa
      481 aaacgtcgat ttttcaagat acagcgtgaa ttttcaggaa atgcggtgag catcacatca
      541 ccacaattca gcaaattgtg aacatcatca cgttcatctt tccctggttg ccaatggccc
      601 attttcctgt cagtaacgag aaggtcgcga attcaggcgc tttttagact ggtcgtaatg
      661 aaattcagca ggatcacata tggatcccaa gcttcttcta gaggtaccgc atgcgatatc
      721 gagctctccc gggaattcca caaattgtta tccgctcaca attccacatg tggaattcca
      781 catgtggaat tcccatgtca gccgttaagt gttcctgtgt cactcaaaat tgctttgaga
      841 ggctctaagg gcttctcagt gtgttacatc cctggcttgt tgtccacaac cgttaaacct
      901 taaaagcttt aaaagcctta tatattcttt tttttcttat aaaacttaaa accttagagg
      961 ctatttaagt tgctgattta tattaatttt attgttcaaa catgagagct tagtacgtga
     1021 aacatgagag cttagtacgt tagccatgag agcttagtac gttagccatg agggtttagt
     1081 tcgttaaaca tgagagctta gtacgttaaa catgagagct tagtacgtga aacatgagag
     1141 cttagtacgt actatcaaca ggttgaactg ctgatcttca gatcctctac gccggacgca
     1201 tcgtggccgg atccagccga ccaggctttc cacgcccgcg tgccgctcca tgtcgttcgc
     1261 gcggttctcg gaaacgcgct gccgcgtttc gtgattgtca cgctcaagcc cgtagtcccg
     1321 ttcgagcgtc gcgcagaggt cagcgagggc gcggtaggcc cgatacggct catggatggt
     1381 gtttcgggtc gggtgaatct tgttgatggc gatatggatg tgcaggttgt cggtgtcgtg
     1441 atgcacggca ctgacgcgct gatgctcggc gaagccaagc ccagcgcaga tgcggtcctc
     1501 aatcgcgcgc aacgtctccg cgtcgggctt ctctcccgcg cggaagctaa ccagcaggtg
     1561 ataggtcttg tcggcctcgg aacgggtgtt gccgtgctgg gtcgccatca cctcggccat
     1621 gacagcgggc agggtgtttg cctcgcagtt cgtgacgcgc acgtgaccca ggcgctcggt
     1681 cttgccttgc tcgtcggtga tgtacttcac cagctccgcg aagtcgctct tcttgatgga
     1741 gcgcatgggg acgtgcttgg caatcacgcg caccccccgg ccgttttagc ggctaaaaaa
     1801 gtcatggctc tgccctcggg cggaccacgc ccatcatgac cttgccaagc tcgtcctgct
     1861 tctcttcgat cttcgccagc agggcgagga tcgtggcatc accgaaccgc gccgtgcgcg
     1921 ggtcgtcggt gagccagagt ttcagcaggc cgcccaggcg gcccaggtcg ccattgatgc
     1981 gggccagctc gcggacgtgc tcatagtcca cgacgcccgt gattttgtag ccctggccga
     2041 cggccagcag gtaggccgac aggctcatgc cggccgccgc cgccttttcc tcaatcgctc
     2101 ttcgttcgtc tggaaggcag tacaccttga taggtgggct gcccttcctg gttggcttgg
     2161 tttcatcagc catccgcttg ccctcatctg ttacgccggc ggtagccggc cagcctcgca
     2221 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc
     2281 cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag
     2341 gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata
     2401 ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaag cgctgcttcc
     2461 ctgctgtttt gtggaatatc taccgactgg aaacaggcaa atgcaggaaa ttactgaact
     2521 gaggggacag gcgagagacg atgccaaaga gctacaccga cgagctggcc gagtgggttg
     2581 aatcccgcgc ggccaagaag cgccggcgtg atgaggctgc ggttgcgttc ctggcggtga
     2641 gggcggatgt cgaggcggcg ttagcgtccg gctatgcgct cgtcaccatt tgggagcaca
     2701 tgcgggaaac ggggaaggtc aagttctcct acgagacgtt ccgctcgcac gccaggcggc
     2761 acatcaaggc caagcccgcc gatgtgcccg caccgcaggc caaggctgcg gaacccgcgc
     2821 cggcacccaa gacgccggag ccacggcggc cgaagcaggg gggcaaggct gaaaagccgg
     2881 cccccgctgc ggccccgacc ggcttcacct tcaacccaac accggacaaa aaggatcctc
     2941 tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat
     3001 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt
     3061 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg
     3121 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     3181 ggtcacagct tgtctgtaag cggatgccgg gagcaggcaa gcccgtcagg gcgcgtcagc
     3241 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     3301 tactggctta actatgcggc atcagagcag attgtactga gactcgagac ggggtctgac
     3361 gctcagtgga acgaaatcga tgataagctg tcaaacatga gcagatcctc tacgccggac
     3421 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
     3481 tcaccgatgg gagatcctaa gatatcgctt aggccacacg ttcaagtgca gccacaggat
     3541 aaatttgcac tgagcctggg tgggattcgg actcgaccgc atagccttca ggagtgagtt
     3601 ttgtgcaata ccaaccgacg acttgaccct gccaagcggc accagatttc ttgcgtacgc
     3661 gatcccctaa gccaaaggtg gcactcaggg gaagcgcaaa ctgccctgca acgggagcgt
     3721 tggcttcatc gctactttga cccatgtcga atccttcttg tgaatctatt atggcgacaa
     3781 gcaaatcgag ctctgactgc ctaccccaca acaactatca gaaagcacca gcacaacggc
     3841 tgcctaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctccataaca
     3901 tcaaacatcg acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta
     3961 caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt
     4021 cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta cttgcattac agtttacgaa
     4081 ccgaacaggc ttatgtcaac tgggttcgtg cgagctcatc gatgcattta atctttctgc
     4141 gaattgagat gacgccactg gctgggcgtc atcccggttt cccgggtaaa caccaccgaa
     4201 aaatagttac tatcttcaaa gccacattcg gtcgaaatat cactgattaa caggcggcta
     4261 tgctggagaa gatattgcgc atgacacact ctgacctgtc gcagatattg attgatggtc
     4321 attccagtct gctggcgaaa ttgctgacgc aaaacgcgct cactgcacga tgcctcatca
     4381 caaaatttat ccagcgcaaa gggacttttc aggctagccg ccagccgggt aatcagctta
     4441 tccagcaacg tttcgctgga tgttggcggc aacgaatcac tggtgtaacg atggcgattc
     4501 agcaacatca ccaactgccc gaacagcaac tcagccattt cgttagcaaa cggcacatgc
     4561 tgactacttt catgctcaag ctgaccgata acctgccgcg cctgcgccat ccccatgcta
     4621 cctaagcgcc agtgtggttg ccctgcgctg gcgttaaatc ccggaatcgc cccctgccag
     4681 tcaagattca gcttcagacg ctccgggcaa taaataatat tctgcaaaac cagatcgtta
     4741 acggaagcgt aggagtgttt atcgtcagca tgaatgtaaa agagatcgcc acgggtaatg
     4801 cgataagggc gatcgttgag tacatgcagg ccattaccgc gccagacaat caccagctca
     4861 caaaaatcat gtgtatgttc agcaaagaca tcttgcggat aacggtcagc cacagcgact
     4921 gcctgctggt cgctggcaaa aaaatcatct ttgagaagtt ttaactgatg cgccaccgtg
     4981 gctacctcgg ccagagaacg aagttgatta ttcgcaatat ggcgtacaaa tacgttgaga
     5041 agattcgcgt tattgcagaa agccatcccg tccctggcga atatcacgcg gtgaccagtt
     5101 aaactctcgg cgaaaaagcg tcgaaaagtg gttactgtcg ctgaatccac agcgataggc
     5161 gatgtcagta acgctggcct cgctgtggcg tagcagatgt cgggctttca tcagtcgcag
     5221 gcggttcagg tatcgctgag gcgtcagtcc cgtttgctgc ttaagctgcc gatgtagcgt
     5281 acgcagtgaa agagaaaatt gatccgccac ggcatcccaa ttcacctcat cggcaaaatg
     5341 gtcctccagc caggccagaa gcaagttgag acgtgatgcg ctgttttcca ggttctcctg
     5401 caaactgctt ttacgcagca agagcagtaa ttgcataaac aagatctcgc gactggcggt
     5461 cgagggtaaa tcattttccc cttcctgctg ttc
//
