LOCUS       sequence-325696-        7543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-range bacterial expression vector with modified 
            rhamnose-inducible promoter. 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 7543)
  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 7543)
  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: pSCrhaB2plus
FEATURES             Location/Qualifiers
     source          1..7543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..772
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             773..1435
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     terminator      complement(1901..1928)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2020..2106)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2259..2276
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             2673..3509
                     /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"
     CDS             3583..4431
                     /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(4460..4465,4466..4482,4483..4488)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4814..5050
                     /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     5074..5093
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5963..5982
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6049..6151)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6268..6286)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcggccaccg gctggctcgc ttcgctcggc ccgtggacaa ccctgctgga caagctgatg
       61 gacaggctgc gcctgcccac gagcttgacc acagggattg cccaccggct acccagcctt
      121 cgaccacata cccaccggct ccaactgcgc ggcctgcggc cttgccccat caattttttt
      181 aattttctct ggggaaaagc ctccggcctg cggcctgcgc gcttcgcttg ccggttggac
      241 accaagtgga aggcgggtca aggctcgcgc agcgaccgcg cagcggcttg gccttgacgc
      301 gcctggaacg acccaagcct atgcgagtgg gggcagtcga agggcgaagc ccgcccgcct
      361 gccccccgag cctcacggcg gcgagtgcgg gggttccaag ggggcagcgc caccttgggc
      421 aaggccgaag gccgcgcagt cgatcaacaa gccccggagg ggccactttt tgccggaggg
      481 ggagccgcgc cgaaggcgtg ggggaacccc gcaggggtgc ccttctttgg gcaccaaaga
      541 actagatata gggcgaaatg cgaaagactt aaaaatcaac aacttaaaaa aggggggtac
      601 gcaacagctc attgcggcac cccccgcaat agctcattgc gtaggttaaa gaaaatctgt
      661 aattgactgc cacttttacg caacgcataa ttgttgtcgc gctgccgaaa agttgcagct
      721 gattgcgcat ggtgccgcaa ccgtgcggca cccctaccgc atggagataa gcatggccac
      781 gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga
      841 acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct
      901 gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac
      961 actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt
     1021 ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt
     1081 ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag
     1141 tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct
     1201 gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga
     1261 gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac
     1321 ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct
     1381 ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt
     1441 gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg
     1501 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga
     1561 cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga
     1621 ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg
     1681 gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
     1741 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa
     1801 tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag
     1861 gcgtttaagt cgaccaataa caagagtttg tagaaacgca aaaaggccat ccgtcaggat
     1921 ggccttctgc ttaatttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     1981 gggccgttgc ttcgcaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
     2041 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag cctttcgttt
     2101 tatttgatgc ctggcagttc cctactctcg catggggaga ccccacacta ccatcggcgc
     2161 tacggcgttt cacttctgag ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc
     2221 caggcaaatt ctgttttatc agaccgcttc tgcgttctga tttaatctgt atcaggctga
     2281 aaatcttctc tcatccgcca aaacagccaa gcttgcatgc ctgcaggtcg actctagagg
     2341 atccccgggt accatggcat atgtgatcct gctgaatttc actagaggga aaccgttgtg
     2401 gtctccctta cgaccagtct aaaaagcgcc tgaattcgcg acctttccct ggttgccagg
     2461 aaaatgggcc attggcaacc agggaaagat gaacgtgatg atgttcacaa tttgctgaat
     2521 tgtggtgatg tgatgctcac cgcatttcct gaaaattcac gctgtatctt gaaaaatcga
     2581 cgttttttac gtggttttcc gtcgaaaatt taaggtaaga acctgacctc gtgattacta
     2641 tttcgccgtg ttgacgacat caggaggcca gtatgaccgt attacatagt gtggattttt
     2701 ttccgtctgg taacgcgtcc gtggcgatag aaccccggct cccgcaggcg gattttcctg
     2761 aacatcatca tgattttcat gaaattgtga ttgtcgaaca tggcacgggt attcatgtgt
     2821 ttaatgggca gccctatacc atcaccggtg gcacggtctg tttcgtacgc gatcatgatc
     2881 ggcatctgta tgaacatacc gataatctgt gtctgaccaa tgtgctgtat cgctcgccgg
     2941 atcgatttca gtttctcgcc gggctgaatc agttgctgcc acaagagctg gatgggcagt
     3001 atccgtctca ctggcgcgtt aaccacagcg tattgcagca ggtgcgacag ctggttgcac
     3061 agatggaaca gcaggaaggg gaaaatgatt taccctcgac cgccagtcgc gagatcttgt
     3121 ttatgcaatt actgctcttg ctgcgtaaaa gcagtttgca ggagaacctg gaaaacagcg
     3181 catcacgtct caacttgctt ctggcctggc tggaggacca ttttgccgat gaggtgaatt
     3241 gggatgccgt ggcggatcaa ttttctcttt cactgcgtac gctacatcgg cagcttaagc
     3301 agcaaacggg actgacgcct cagcgatacc tgaaccgcct gcgactgatg aaagcccgac
     3361 atctgctacg ccacagcgag gccagcgtta ctgacatcgc ctatcgctgt ggattcagcg
     3421 acagtaacca cttttcgacg ctttttcgcc gagagtttaa ctggtcaccg cgtgatattc
     3481 gccagggacg ggatggcttt ctgcaataac gcgaatcttc tcaacgtatt tgtacgccat
     3541 attgcgaata atcaacttcg ttctctggcc gaggtagcca cggtggcgca tcagttaaaa
     3601 cttctcaaag atgatttttt tgccagcgac cagcaggcag tcgctgtggc tgaccgttat
     3661 ccgcaagatg tctttgctga acatacacat gatttttgtg agctggtgat tgtctggcgc
     3721 ggtaatggcc tgcatgtact caacgatcgc ccttatcgca ttacccgtgg cgatctcttt
     3781 tacattcatg ctgacgataa acactcctac gcttccgtta acgatctggt tttgcagaat
     3841 attatttatt gcccggagcg tctgaagctg aatcttgact ggcagggggc gattccggga
     3901 tttaacgcca gcgcagggca accacactgg cgcttaggta gcatggggat ggcgcaggcg
     3961 cggcaggtta tcggtcagct tgagcatgaa agtagtcagc atgtgccgtt tgctaacgaa
     4021 atggctgagt tgctgttcgg gcagttggtg atgttgctga atcgccatcg ttacaccagt
     4081 gattcgttgc cgccaacatc cagcgaaacg ttgctggata agctgattac ccggctggcg
     4141 gctagcctga aaagtccctt tgcgctggat aaattttgtg atgaggcatc gtgcagtgag
     4201 cgcgttttgc gtcagcaatt tcgccagcag actggaatga ccatcaatca atatctgcga
     4261 caggtcagag tgtgtcatgc gcaatatctt ctccagcata gccgcctgtt aatcagtgat
     4321 atttcgaccg aatgtggctt tgaagatagt aactattttt cggtggtgtt tacccgggaa
     4381 accgggatga cgcccagcca gtggcgtcat ctcaattcgc agaaagatta aatgcatcga
     4441 tgagctcgca cgaacccagt tgacataagc ctgttcggtt cgtaaactgt aatgcaagta
     4501 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
     4561 agtggcggtt ttcatggctt gttatgactg tttttttgta cagtctatgc ctcgggcatc
     4621 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
     4681 tacgcagcag ggcagtcgcc ctaaaacaaa gttaggcagc cgttgtgctg gtgctttctg
     4741 atagttgttg tggggtaggc agtcagagct cgatttgctt gtcgccataa tagattcaca
     4801 agaaggattc gacatgggtc aaagtagcga tgaagccaac gctcccgttg cagggcagtt
     4861 tgcgcttccc ctgagtgcca cctttggctt aggggatcgc gtacgcaaga aatctggtgc
     4921 cgcttggcag ggtcaagtcg tcggttggta ttgcacaaaa cttactcctg aaggctatgc
     4981 ggtcgagtcc gaatcccacc caggctcagt gcaaatttat cctgtggctg cacttgaacg
     5041 tgtggcctaa gcgatatctt aggatctccc atcggtgatg tcggcgatat aggcgccagc
     5101 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatctgctc
     5161 atgtttgaca gcttatcatc gatttcgttc cactgagcgt cagaccccgt agaccactca
     5221 ctacagcaga gccatttaaa caacatcccc tccccctttc caccgcgtca gacgcccgta
     5281 gcagcccgct acgggctttt tcatgccctg ccctagcgtc caagcctcac ggccgcgctc
     5341 ggcctctctg gcggccttct ggcgctgagg tctgcctcgt gaagaaggtg ttgctgactc
     5401 ataccaggcc tgaatcgccc catcatccag ccagaaagtg agggagccac ggttgatgag
     5461 agctttgttg taggtggacc agttggtgat tttgaacttt tgctttgcca cggaacggtc
     5521 tgcgttgtcg ggaagatgcg tgatctgatc cttcaactca gcaaaagttc gatttattca
     5581 acaaagccgc cgtcccgtca agtcagcgta atgctctgcc agtgttacaa ccaattaacc
     5641 aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     5701 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     5761 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     5821 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     5881 gtgacgactg aatccggtga gaatggcaaa agctaaaaag gccgtaatat ccagctgaac
     5941 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     6001 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     6061 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     6121 gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca
     6181 gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt
     6241 cacaggtatt tattcgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     6301 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcgccc
     6361 gcgttcctgc tggcgctggg cctgtttctg gcgctggact tcccgctgtt ccgtcagcag
     6421 cttttcgccc acggccttga tgatcgcggc ggccttggcc tgcatatccc gattcaacgg
     6481 ccccagggcg tccagaacgg gcttcaggcg ctcccgaagg tctcgggccg tctcttgggc
     6541 ttgatcggcc ttcttgcgca tctcacgcgc tcctgcggcg gcctgtaggg caggctcata
     6601 cccctgccga accgcttttg tcagccggtc ggccacggct tccggcgtct caacgcgctt
     6661 tgagattccc agcttttcgg ccaatccctg cggtgcatag gcgcgtggct cgaccgcttg
     6721 cgggctgatg gtgacgtggc ccactggtgg ccgctccagg gcctcgtaga acgcctgaat
     6781 gcgcgtgtga cgtgccttgc tgccctcgat gccccgttgc agccctagat cggccacagc
     6841 ggccgcaaac gtggtctggt cgcgggtcat ctgcgctttg ttgccgatga actccttggc
     6901 cgacagcctg ccgtcctgcg tcagcggcac cacgaacgcg gtcatgtgcg ggctggtttc
     6961 gtcacggtgg atgctggccg tcacgatgcg atccgccccg tacttgtccg ccagccactt
     7021 gtgcgccttc tcgaagaacg ccgcctgctg ttcttggctg gccgacttcc accattccgg
     7081 gctggccgtc atgacgtact cgaccgccaa cacagcgtcc ttgcgccgct tctctggcag
     7141 caactcgcgc agtcggccca tcgcttcatc ggtgctgctg gccgcccagt gctcgttctc
     7201 tggcgtcctg ctggcgtcag cgttgggcgt ctcgcgctcg cggtaggcgt gcttgagact
     7261 ggccgccacg ttgcccattt tcgccagctt cttgcatcgc atgatcgcgt atgccgccat
     7321 gcctgcccct cccttttggt gtccaaccgg ctcgacgggg gcagcgcaag gcggtgcctc
     7381 cggcgggcca ctcaatgctt gagtatactc actagacttt gcttcgcaaa gtcgtgaccg
     7441 cctacggcgg ctgcggcgcc ctacgggctt gctctccggg cttcgccctg cgcggtcgct
     7501 gcgctccctt gccagcccgt ggatatgtgg acgatggccg cga
//
