LOCUS       Exported                4745 bp ds-DNA    circular SYN 15-11-2014
DEFINITION  pUC18-derived plasmid encoding the rhaB promoter plus rhaR and rhaS 
            genes, for rhamnose-inducible protein expression in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    pRHA-67
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4745)
  AUTHORS   Giacalone MJ, Gentile AM, Lovitt BT, Berkley NL, Gunderson CW, 
            Surber MW.
  TITLE     Toxic protein expression in Escherichia coli using a rhamnose-based 
            tightly regulated and tunable promoter system.
  JOURNAL   BioTechniques 2006;40:355-64.
  PUBMED    16568824
REFERENCE   2  (bases 1 to 4745)
  AUTHORS   Xbrane Bioscience
  TITLE     Direct Submission
COMMENT     This plasmid sequence was reconstructed from the procedure described
            in the original paper. The plasmid size is slightly different than 
            reported.
FEATURES             Location/Qualifiers
     source          1..4745
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     gene            405..410
                     /gene="rhaT"
                     /gene_synonym="ECK3900; JW3878"
                     /locus_tag="b3907"
                     /note="rhaT"
                     /db_xref="EcoGene:EG11313"
                     /db_xref="GeneID:948398"
     CDS             complement(407..1255)
                     /codon_start=1
                     /transl_table=11
                     /gene="rhaR"
                     /gene_synonym="ECK3899; JW3877; rhaC1"
                     /locus_tag="b3906"
                     /product="transcriptional activator of rhaSR"
                     /function="regulator; Degradation of small molecules: 
                     Carbon compounds"
                     /note="rhaR"
                     /note="positive regulator for rhaRS operon"
                     /db_xref="GI:171701684"
                     /db_xref="ASAP:ABE-0012747"
                     /db_xref="UniProtKB/Swiss-Prot:P09378"
                     /db_xref="EcoGene:EG10842"
                     /db_xref="GeneID:948396"
                     /protein_id="NP_418342.2"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     gene            complement(407..1255)
                     /gene="rhaR"
                     /gene_synonym="ECK3899; JW3877; rhaC1"
                     /locus_tag="b3906"
                     /note="rhaR"
                     /db_xref="EcoGene:EG10842"
                     /db_xref="GeneID:948396"
     CDS             complement(1329..2165)
                     /codon_start=1
                     /transl_table=11
                     /gene="rhaS"
                     /gene_synonym="ECK3898; JW3876; rhaC2"
                     /locus_tag="b3905"
                     /product="transcriptional activator of rhaBAD and rhaT"
                     /function="regulator; Degradation of small molecules: 
                     Carbon compounds"
                     /note="rhaS"
                     /note="positive regulator for rhaBAD operon"
                     /db_xref="GI:16131745"
                     /db_xref="ASAP:ABE-0012745"
                     /db_xref="UniProtKB/Swiss-Prot:P09377"
                     /db_xref="EcoGene:EG10843"
                     /db_xref="GeneID:948397"
                     /protein_id="NP_418341.1"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     gene            complement(1329..2165)
                     /gene="rhaS"
                     /gene_synonym="ECK3898; JW3876; rhaC2"
                     /locus_tag="b3905"
                     /note="rhaS"
                     /db_xref="EcoGene:EG10843"
                     /db_xref="GeneID:948397"
     promoter        2311..2429
                     /note="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)"
     misc_feature    2439..2444
                     /note="RBS"
                     /note="ribosome binding site"
     misc_feature    2447..2508
                     /gene="
                     "
                     /note="MCS"
                     /note="pUC19 multiple cloning site"
     primer_bind     complement(2524..2540)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2548..2564
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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)."
     promoter        complement(2572..2602)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(2926..3514)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3685..4545)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4546..4650)
                     /gene="bla"
                     /note="AmpR promoter"