pCEV-G3-Km vector (V008565)

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LOCUS       40924_10426        6645 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning and expression vector pCEV-G3-Km, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6645)
  AUTHORS   Vickers CE, Bydder SF, Zhou Y, Nielsen LK.
  TITLE     Dual gene expression cassette vectors with antibiotic selection 
            markers for engineering in Saccharomyces cerevisiae
  JOURNAL   Microb. Cell Fact. 12 (1), 96 (2013)
  PUBMED    24161108
REFERENCE   2  (bases 1 to 6645)
  AUTHORS   Bydder SF, Vickers CE.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JUL-2013) AIBN, The University of Queensland, Crn 
            College and Cooper Roads (Bldg 75), St Lucia, QLD 4072, Australia
REFERENCE   3  (bases 1 to 6645)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6645)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb. 
            Cell Fact."; date: "2013"; volume: "12"; issue: "1"; pages: "96"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-JUL-2013) AIBN, The University of Queensland, Crn College and 
            Cooper Roads (Bldg 75), St Lucia, QLD 4072, Australia"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(2..167)
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(315..338)
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     promoter        356..1008
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     promoter        1017..1424
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1444..1462
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1480..1509
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     terminator      1539..1728
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(1971..2559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2733..3590)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3591..3695)
                     /label=AmpR promoter
     rep_origin      complement(3722..5064)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    5084..5117
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            5172..6528
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     misc_recomb     6591..6624
                     /label=loxP recognition sequence for Cre recombinase
                     /note="loxP recognition sequence for Cre recombinase"
     protein_bind    complement(6591..6624)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."