LOCUS       Exported                5654 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSKPDR5-PPUS, complete sequence.
ACCESSION   JN581374
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5654)
  AUTHORS   Nakamura K, Niimi M, Niimi K, Holmes AR, Yates JE, Decottignies A, 
            Monk BC, Goffeau A, Cannon RD.
  TITLE     Functional expression of Candida albicans drug efflux pump Cdr1p in 
            a Saccharomyces cerevisiae strain deficient in membrane transporters
  JOURNAL   Antimicrob. Agents Chemother. 45 (12), 3366-3374 (2001)
  PUBMED    11709310
REFERENCE   2  (bases 1 to 5654)
  AUTHORS   Lamping E, Niimi M, Cannon RD.
  TITLE     Small, synthetic, GC-rich mRNA stem-loop modules 5 proximal to the 
            AUG start-codon predictably tune gene expression in yeast
  JOURNAL   Microb. Cell Fact. 12 (1), 74 (2013)
  PUBMED    23895661
REFERENCE   3  (bases 1 to 5654)
  AUTHORS   Lamping E, Niimi M, Cannon RD.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-AUG-2011) Sir John Walsh Research Institute, 
            University of Otago, 310 Great King Street, Dunedin, Otago 9054, New
            Zealand
REFERENCE   4  (bases 1 to 5654)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5654
                     /organism="Cloning vector pSKPDR5-PPUS"
                     /mol_type="other DNA"
                     /label=pSK-PDR5-PPUS
                     /note="pSK-PDR5-PPUS"
                     /db_xref="taxon:1093921"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    650..3453
                     /label=Saccharomyces cerevisiae transformation cassette
                     /note="Saccharomyces cerevisiae transformation cassette"
     regulatory      650..1822
                     /regulatory_class="promoter"
                     /gene="PDR5"
                     /note="Saccharomyces cerevisiae PDR5 promoter"
     gene            650..1822
                     /gene="PDR5"
                     /label=PDR5
     primer_bind     667..683
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1823..1864
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      1870..2069
                     /regulatory_class="terminator"
                     /note="3 end of Saccharomyces cerevisiae PDR5 ORF"
     misc_feature    2075..3145
                     /label=URA3 selection marker
                     /note="URA3 selection marker"
     promoter        2106..2321
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2322..3125
                     /codon_start=1
                     /gene="URA3"
                     /product="orotidine-5-phosphate decarboxylase"
                     /label=URA3
                     /note="Ura3p; OMP decarboxylase"
                     /protein_id="AEQ55255.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            2322..3125
                     /gene="URA3"
                     /label=URA3
     misc_feature    3146..3423
                     /label=3 end of Saccharomyces cerevisiae PDR5 ORF
                     /note="3 end of Saccharomyces cerevisiae PDR5 ORF"
     promoter        complement(3466..3484)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3505..3521)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3529..3545
                     /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(3553..3583)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3598..3619
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3907..4495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4666..5526)
                     /codon_start=1
                     /gene="Ap"
                     /product="beta-lactamase"
                     /label=Ap
                     /note="E. coli ampicillin resistance protein"
                     /protein_id="AEQ55256.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4666..5526)
                     /gene="Ap"
                     /label=Ap
     promoter        complement(5527..5631)
                     /gene="bla"
                     /label=AmpR promoter