LOCUS       Exported                6880 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pRS446PGK DNA, complete sequence.
ACCESSION   AB304875
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6880)
  AUTHORS   Mumberg D, Muller R, Funk M.
  TITLE     Yeast vectors for the controlled expression of heterologous proteins
            in different genetic backgrounds
  JOURNAL   Gene 156 (1), 119-122 (1995)
  PUBMED    7737504
REFERENCE   2  (bases 1 to 6880)
  AUTHORS   Ohto C, Muramatsu M, Obata S, Sakuradani E, Shimizu S.
  TITLE     Overexpression of the gene encoding HMG-CoA reductase in 
            Saccharomyces cerevisiae for production of prenyl alcohols
  JOURNAL   Appl. Microbiol. Biotechnol. 82 (5), 837-845 (2009)
  PUBMED    19083230
REFERENCE   3  (bases 1 to 6880)
  AUTHORS   Ohto C.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-MAY-2007) Contact:Chikara Ohto Toyota Motor 
            Corporation, Bio Research Laboratory, Future Project Division; 1, 
            Toyota-cho, Toyota, Aichi 471-8572, Japan
REFERENCE   4  (bases 1 to 6880)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6880
                     /organism="Shuttle vector pRS446PGK"
                     /mol_type="other DNA"
                     /note="Saccharomyces cerevisiae - Escherichia coli 
                     expression shuttle vector"
                     /db_xref="taxon:446215"
     promoter        201..416
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             417..1220
                     /codon_start=1
                     /gene="URA3"
                     /product="orotidine-5-phosphate decarboxylase"
                     /label=URA3
                     /protein_id="BAG16809.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            417..1220
                     /gene="URA3"
                     /label=URA3
     rep_origin      2270..3150
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     3487..3503
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3513..3531
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(3540..3805)
                     /regulatory_class="terminator"
                     /gene="CYC1"
     gene            complement(3540..3805)
                     /gene="CYC1"
                     /label=CYC1
     terminator      3550..3797
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3807..3823
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3857..3873)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(3884..4602)
                     /regulatory_class="promoter"
                     /gene="PGK1"
     gene            complement(3884..4602)
                     /gene="PGK1"
                     /label=PGK1
     promoter        complement(4620..4638)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4659..4675)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4683..4699
                     /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(4707..4737)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(5061..5649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5820..6680)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6681..6785)
                     /gene="bla"
                     /label=AmpR promoter