LOCUS       Exported                6345 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector yGALset352, complete sequence.
ACCESSION   AF041803
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6345)
  AUTHORS   Enomoto S, Chen G, Berman J.
  TITLE     Vectors for expressing T7 epitope- and His6 affinity-tagged fusion 
            proteins in S. cerevisiae
  JOURNAL   BioTechniques 24 (5), 782-786 (1998)
  PUBMED    9591127
REFERENCE   2  (bases 1 to 6345)
  AUTHORS   Enomoto S, Berman J.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-JAN-1998) Plant Biology, University of Minnesota, 220 
            Biological Sciences Center, St. Paul, MN 55108, USA
REFERENCE   3  (bases 1 to 6345)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6345
                     /organism="Cloning vector yGALset352"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:71550"
     promoter        70..88
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      112..793
                     /regulatory_class="promoter"
                     /note="S. cerevisiae GAL10 promoter"
     promoter        121..785
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    452..569
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     misc_feature    794..929
                     /label=5 recombination region
                     /note="5 recombination region"
     RBS             804..826
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    846..863
                     /label=polyhistidine affinity tag
                     /note="polyhistidine affinity tag"
     CDS             846..863
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    867..899
                     /label=T7 tag epitope
                     /note="T7 tag epitope"
     CDS             867..899
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             903..926
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      912..926
                     /regulatory_class="other"
                     /note="enterokinase recognition sequence"
     misc_feature    936..982
                     /label=multiple cloning sites
                     /note="multiple cloning sites"
     misc_feature    983..1188
                     /label=3 recombination region
                     /note="3 recombination region"
     terminator      1046..1093
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     gene            1247..2099
                     /gene="cen4"
                     /label=cen4
                     /note="S. cerevisiae CEN4"
     misc_feature    1807..2969
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      2128..3218
                     /label=S. cerevisiae ARS1
                     /note="S. cerevisiae ARS1"
     rep_origin      2132..2969
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     gene            complement(3226..4383)
                     /gene="ura3"
                     /label=ura3
                     /note="S. cerevisiae URA3"
     CDS             complement(3304..4107)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4108..4323)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     promoter        4439..4543
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4544..5404
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicillin resistance gene (apr/amp); E. coli 
                     beta-lactamase gene"
                     /protein_id="AAC40218.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5575..6163
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      6157
                     /label=pUC origin
                     /note="pUC origin"