Basic Vector Information
YEplac112 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 ℃.
YEplac112 vector Sequence
LOCUS 40924_49652 4989 bp DNA circular SYN 01-JAN-1980 DEFINITION Yeast episomal plasmid with a TRP1 marker. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4989) AUTHORS Gietz RD, Sugino A. TITLE New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. JOURNAL Gene 1988;74:527-34. PUBMED 3073106 REFERENCE 2 (bases 1 to 4989) TITLE Direct Submission REFERENCE 3 (bases 1 to 4989) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; date: "1988"; volume: "74"; pages: "527-34" COMMENT SGRef: number: 2; type: "Journal Article" COMMENT The sequence of this plasmid was reconstructed using the description from Gietz and Sugino. FEATURES Location/Qualifiers source 1..4989 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 106..127 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 142..172 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 180..196 /label=lac operator /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 204..220 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" misc_feature complement(233..289) /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(290..306) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin complement(743..2089) /direction=LEFT /label=2u ori /note="yeast 2u plasmid origin of replication" CDS complement(2204..2875) /codon_start=1 /label=TRP1 /note="phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis" /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA KK" promoter complement(2876..2977) /label=TRP1 promoter promoter 3083..3187 /label=AmpR promoter CDS 3188..4045 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 4219..4807 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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