LOCUS       Exported                7357 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pVTLU260, complete sequence.
ACCESSION   AY974039
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7357)
  AUTHORS   Mok CSL., Melcher K.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-MAR-2005) School of Biomedical Sciences, University of
            Ulster, Cromore Road, Coleraine BT52 1SA, UK
REFERENCE   2  (bases 1 to 7357)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7357
                     /organism="Expression vector pVTLU260"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
                     /db_xref="taxon:322728"
     promoter        29..244
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             245..1048
                     /codon_start=1
                     /gene="URA3-deltaNcoI"
                     /product="orotidine-5-phosphate decarboxylate"
                     /label=URA3-deltaNcoI
                     /note="Ura3p; NcoI site removed by site-specific 
                     mutagenesis without altering amino acid sequence"
                     /protein_id="AAX85440.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            245..1048
                     /gene="URA3-deltaNcoI"
                     /label=URA3-deltaNcoI
     promoter        1246..1350
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1351..2211
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDVGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             1900..2211
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /note="beta-lactamase; antibiotic resistance marker"
                     /protein_id="AAX85441.1"
                     /translation="MATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWF
                     IADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     gene            1900..2211
                     /gene="ampR"
                     /label=ampR
     rep_origin      2382..2970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      2694
                     /label=Escherichia coli replication initiation base
                     /note="Escherichia coli replication initiation base"
     regulatory      2999..3400
                     /regulatory_class="promoter"
                     /note="ADH1 promoter"
     promoter        2999..3400
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             3409..3429
                     /codon_start=1
                     /product="His6-tag"
                     /label=His6-tag
                     /note="affinity purification tag"
                     /protein_id="AAX85442.1"
                     /translation="MHHHHHH"
     CDS             3412..3429
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    3436..3456
                     /note="cleavage site for Tobacco Etch Virus (TEV) protease;
                     allows site-specific proteolytic cleavage of His6-GST 
                     tandem affinity tags"
     CDS             3436..3456
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     regulatory      3639..3831
                     /regulatory_class="terminator"
                     /note="ADH1 terminator"
     terminator      3639..3826
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     rep_origin      3837..5183
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      3883..5183
                     /label=from yeast endogenous 2 micron plasmid
                     /note="from yeast endogenous 2 micron plasmid"
     CDS             5557..6651
                     /codon_start=1
                     /gene="LEU2"
                     /allele="leu2d"
                     /product="beta-isopropylmalate dehydrogenase"
                     /label=LEU2
                     /note="Leu2p"
                     /protein_id="AAX85443.1"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEALRYPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPGFAKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     gene            5557..6651
                     /gene="LEU2"
                     /allele="leu2d"
                     /label=LEU2
     primer_bind     7319..7335
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"