pSKPDR5-PGK1 vector (V003153)

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LOCUS       40924_40592        5773 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSKPDR5-PGK1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5773)
  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   2  (bases 1 to 5773)
  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   3  (bases 1 to 5773)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5773)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb. 
            Cell Fact."; date: "2013"; volume: "12"; issue: "1"; pages: "74"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-AUG-2011) Sir John Walsh Research Institute, University of 
            Otago, 310 Great King Street, Dunedin, Otago 9054, New Zealand"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     primer_bind     667..683
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1919..2189
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate
                     kinase gene"
     promoter        2225..2440
                     /label=URA3 promoter
     CDS             2441..3241
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     misc_feature    3265..3542
                     /label=3' end of Saccharomyces cerevisiae PDR5 ORF
                     /note="3' end of Saccharomyces cerevisiae PDR5 ORF"
     promoter        complement(3585..3603)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3624..3640)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3648..3664)
                     /label=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(3672..3702)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3717..3738)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4026..4614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4788..5645)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5646..5750)
                     /label=AmpR promoter