LOCUS       Exported                7518 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector pSCrhaB2, complete sequence.
ACCESSION   AY917125
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7518)
  AUTHORS   Cardona ST, Valvano MA.
  TITLE     An expression vector containing a rhamnose-inducible promoter 
            provides tightly regulated gene expression in Burkholderia 
            cenocepacia
  JOURNAL   Plasmid 54 (3), 219-228 (2005)
  PUBMED    15925406
REFERENCE   2  (bases 1 to 7518)
  AUTHORS   Cardona ST, Valvano MA.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-FEB-2005) Microbiology & Immunology, University of 
            Western Ontario, DSB 3014, London, Ontario N6A 5C1, Canada
REFERENCE   3  (bases 1 to 7518)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7518
                     /organism="Broad host range vector pSCrhaB2"
                     /mol_type="other DNA"
                     /db_xref="taxon:324138"
     CDS             complement(164..400)
                     /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(727..755)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             complement(786..1808)
                     /codon_start=1
                     /gene="rhaR"
                     /product="RhaR"
                     /label=rhaR
                     /note="transcriptional regulator"
                     /protein_id="AAX99114.1"
                     /translation="MDSATVTTFRRFFAESLTGHRVIFARDGMAFCNNANLLNVFVRHI
                     ANNQLRSLAEVATVAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVWR
                     GNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAIP
                     GFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHRY
                     TSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTINQ
                     YLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQKD
                     "
     gene            complement(786..1808)
                     /gene="rhaR"
                     /label=rhaR
     CDS             complement(786..1634)
                     /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(1708..2544)
                     /codon_start=1
                     /gene="rhaS"
                     /product="RhaS"
                     /label=rhaS
                     /note="transcriptional regulator"
                     /protein_id="AAX99113.1"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     gene            complement(1708..2544)
                     /gene="rhaS"
                     /label=rhaS
     promoter        2690..2808
                     /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    2701..2730
                     /label=CRP binding site
                     /bound_moiety="CRP"
     regulatory      2821..2824
                     /regulatory_class="ribosome_binding_site"
     misc_feature    2828..2882
                     /label=multi-cloning site
                     /note="multi-cloning site"
     terminator      3085..3171
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3263..3290
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        complement(4145..4247)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      5640..6409
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             6410..7072
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"