Basic Vector Information
YEp85 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 ℃.
YEp85 vector Sequence
LOCUS 40924_49647 6801 bp DNA circular SYN 18-DEC-2018 DEFINITION Episomal vector YEp85, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6801) AUTHORS Baruffini E, Serafini F, Lodi T. TITLE Construction and characterization of centromeric, episomal and GFP-containing vectors for Saccharomyces cerevisiae prototrophic strains JOURNAL J. Biotechnol. 143 (4), 247-254 (2009) PUBMED 19683551 REFERENCE 2 (bases 1 to 6801) AUTHORS Baruffini E, Serafini F, Lodi T. TITLE Direct Submission JOURNAL Submitted (18-MAR-2009) Department of Genetics, Biology of Microorganisms, Anthropology, Evolution, University of Parma, Viale Usberti 11/A, Parma 43100, Italy REFERENCE 3 (bases 1 to 6801) TITLE Direct Submission REFERENCE 4 (bases 1 to 6801) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J. Biotechnol."; date: "2009"; volume: "143"; issue: "4"; pages: "247-254" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (18-MAR-2009) Department of Genetics, Biology of Microorganisms, Anthropology, Evolution, University of Parma, Viale Usberti 11/A, Parma 43100, Italy" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..6801 /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" gene 2243..3599 /label=kanMX /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" promoter complement(4392..4789) /label=LEU2 promoter promoter 4895..4999 /label=AmpR promoter CDS 5000..5857 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 6031..6619 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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