Basic Vector Information
yGALset983 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
yGALset983 vector Sequence
LOCUS 40924_49712 6483 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector yGALset983, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6483) 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 6483) 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 6483) TITLE Direct Submission REFERENCE 4 (bases 1 to 6483) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "BioTechniques"; date: "1998"; volume: "24"; issue: "5"; pages: "782-786" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (09-JAN-1998) Plant Biology, University of Minnesota, 220 Biological Sciences Center, St. Paul, MN 55108, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..6483 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 70..88 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(121..785) /label=GAL1,10 promoter /note="divergent inducible promoter, regulated by Gal4" RBS 804..826 /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 846..863 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" CDS 867..899 /codon_start=1 /label=T7 tag (gene 10 leader) /note="leader peptide from bacteriophage T7 gene 10" /translation="MASMTGGQQMG" CDS 903..926 /codon_start=1 /label=Xpress(TM) tag /note="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /translation="DLYDDDDK" misc_feature 936..982 /label=multiple cloning sites /note="multiple cloning sites" terminator 1046..1093 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin complement(1485..2831) /direction=LEFT /label=2u ori /note="yeast 2u plasmid origin of replication" CDS complement(2972..4012) /codon_start=1 /label=LEU2 /note="3-isopropylmalate dehydrogenase, required for leucine biosynthesis" /translation="KITAEAIKVLKAISDVRSNVKFDFENHLIGGAAIDATGVPLPDEA LEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYANLRPCNFASDSLLDLSP IKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYTVPEVQRITRMAAFMALQ HEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQLIDSAAMILVKNPTHLN GIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAFGLYEPCHGSAPDLPKNK VDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTGDLGGSNSTTEVGDAVAE EVKKILA" promoter complement(4074..4471) /label=LEU2 promoter promoter 4577..4681 /label=AmpR promoter CDS 4682..5539 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 5713..6301 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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