LOCUS       Exported                6558 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCEV-G2-Ph, complete sequence.
ACCESSION   KF154123
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6558)
  AUTHORS   Behrendorff JB, Vickers CE, Chrysanthopoulos P, Nielsen LK.
  TITLE     2,2-Diphenyl-1-picrylhydrazyl as a screening tool for recombinant 
            monoterpene biosynthesis
  JOURNAL   Microb. Cell Fact. 12 (1), 76 (2013)
  PUBMED    23968454
REFERENCE   2  (bases 1 to 6558)
  AUTHORS   Vickers CE.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-MAY-2013) Australian Institute for Bioengineering and 
            Nanotechnology, The University of Queensland, Cnr College and Cooper
            Rds, St Lucia, QLD 4072, Australia
REFERENCE   3  (bases 1 to 6558)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6558
                     /organism="Expression vector pCEV-G2-Ph"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
                     /db_xref="taxon:1399550"
     terminator      2..167
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     regulatory      complement(2..166)
                     /regulatory_class="terminator"
                     /note="Saccharomyces cerevisiae ADH1 terminator"
     misc_feature    complement(315..341)
                     /label=FLAG tag
                     /note="FLAG tag"
     CDS             complement(315..338)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      complement(360..1343)
                     /regulatory_class="promoter"
                     /note="Saccharomyces cerevisiae PGK1 promoter"
     regulatory      1353..1772
                     /regulatory_class="promoter"
                     /note="Saccharomyces cerevisiae TEF1 promoter"
     promoter        1354..1760
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1780..1798
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1813..1848
                     /label=c-myc tag
                     /note="c-myc tag"
     CDS             1816..1845
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      1875..2064
                     /regulatory_class="terminator"
                     /note="Saccharomyces cerevisiae CYC1 terminator"
     terminator      1875..2064
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      2251..2918
                     /label=pUC ori
                     /note="pUC ori"
     rep_origin      complement(2307..2895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3066..3926)
                     /codon_start=1
                     /gene="bla"
                     /product="beta lactamase"
                     /label=bla
                     /note="confers ampicillin resistance"
                     /protein_id="AGV68805.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3066..3926)
                     /gene="bla"
                     /label=bla
     promoter        complement(3927..4031)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4058..5400
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      4060..5206
                     /label=2mu Saccharomyces cerevisiae origin of replication
                     /note="2mu Saccharomyces cerevisiae origin of replication"
     protein_bind    complement(5021..5068)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_recomb     5420..5453
                     /label=loxP recognition sequence for Cre recombinase
                     /note="loxP recognition sequence for Cre recombinase"
     protein_bind    complement(5420..5453)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      5454..5850
                     /regulatory_class="promoter"
                     /note="Ashbya gossypii TEF promoter"
     promoter        5508..5851
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5852..6238
                     /codon_start=1
                     /gene="ble"
                     /product="bleomycin resistance protein"
                     /label=ble
                     /protein_id="AGV68806.1"
                     /translation="MGMTDQATPNLPSRDFDPTAAFYERLGFGIVFRDAGWMILQRGDL
                     MLEFFAHPGLDPLASWFSCCLRLDDLAEFYRQCKSVGIQETSSGYPRIHAPELQEWGGT
                     MAALVDPDGTLLRLIQNELLAGIS"
     gene            5852..6238
                     /gene="ble"
                     /label=ble
     terminator      6244..6441
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     regulatory      6245..6503
                     /regulatory_class="terminator"
                     /note="Ashbya gossypii TEF terminator"
     misc_recomb     6504..6537
                     /label=loxP recognition sequence for Cre recombinase
                     /note="loxP recognition sequence for Cre recombinase"
     protein_bind    complement(6504..6537)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."