LOCUS       Exported                6709 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Reporter vector pGUSA, complete sequence.
ACCESSION   AY292368
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6709)
  AUTHORS   Girbal L, Mortier-Barriere I, Raynaud F, Rouanet C, Croux C, 
            Soucaille P.
  TITLE     Development of a sensitive gene expression reporter system and an 
            inducible promoter-repressor system for Clostridium acetobutylicum
  JOURNAL   Appl. Environ. Microbiol. 69 (8), 4985-4988 (2003)
  PUBMED    12902297
REFERENCE   2  (bases 1 to 6709)
  AUTHORS   Girbal L.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAY-2003) Laboratoire de Biotechnologie-Bioprocedes, 
            UMR CNRS 5504, UR INRA 792, INSA, 135, avenue de Rangueil, Toulouse 
            31077, France
REFERENCE   3  (bases 1 to 6709)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6709
                     /organism="Reporter vector pGUSA"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:241190"
     misc_feature    2..6672
                     /note="polylinker with PstI, XbaI, KpnI, SmaI, NotI, SacII 
                     restriction sites"
     CDS             20..1828
                     /codon_start=1
                     /gene="gusA"
                     /product="beta-glucuronidase"
                     /label=gusA
                     /note="from Escherichia coli; UidA"
                     /protein_id="AAQ21188.1"
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQVHGNISPLAEATRKLDPTRPITCVNVMFCDAHTDTISDL
                     FDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMYT
                     DMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRKP
                     KSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            20..1828
                     /gene="gusA"
                     /label=gusA
     primer_bind     complement(2008..2024)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2032..2048
                     /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)."
     promoter        complement(2056..2086)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2101..2122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2410..2998)
                     /direction=LEFT
                     /label=pMB1 origin of replication from pUC18
                     /note="pMB1 origin of replication from pUC18"
     CDS             complement(3169..4029)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance protein"
                     /protein_id="AAQ21189.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3169..4029)
                     /gene="bla"
                     /label=bla
     promoter        complement(4030..4134)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4610..4626
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4694..5134)
                     /codon_start=1
                     /gene="repL"
                     /product="RepL"
                     /label=repL
                     /note="Bacillus subtilis plasmid pIM13 replication protein"
                     /protein_id="AAQ21190.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            complement(4694..5134)
                     /gene="repL"
                     /label=repL
     CDS             complement(5582..6316)
                     /codon_start=1
                     /gene="mlsR"
                     /product="MlsR"
                     /label=mlsR
                     /note="Bacillus subtilis plasmid pIM13 
                     macrolide-lincosamide-streptogramin B resistance protein"
                     /protein_id="AAQ21191.1"
                     /translation="MNEKNIKHSQNFITSKHNIDKIMTNIRLNEHDNIFEIGSGKGHFT
                     LELVQRCNFVTAIEIDHKLCKTTENKLVDHDNFQVLNKDILQFKFPKNQSYKIFGNIPY
                     NISTDIIRKIVFDSIADEIYLIVEYGFAKRLLNTKRSLALFLMAEVDISILSMVPREYF
                     HPKPKVNSSLIRLNRKKSRISHKDKQKYNYFVMKWVNKEYKKIFTKNQFNNSLKHAGID
                     DLNNISFEQFLSLFNSYKLFNK"
     gene            complement(5582..6316)
                     /gene="mlsR"
                     /label=mlsR