Basic Vector Information
- Vector Name:
- YEp352
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5181 bp
- Type:
- Yeast Plasmids
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Hill JE, Myers AM, Koerner TJ, Tzagoloff A.
- Copy Number:
- High copy number
- Promoter:
- URA3
- 5' Primer:
- M13 fwd
- 3' Primer:
- M13 rev
YEp352 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 ℃.
YEp352 vector Sequence
LOCUS 40924_49547 5181 bp DNA circular SYN 01-JAN-1980
DEFINITION Yeast/E. coli shuttle vector with a URA3 marker.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5181)
AUTHORS Hill JE, Myers AM, Koerner TJ, Tzagoloff A.
TITLE Yeast/E. coli shuttle vectors with multiple unique restriction
sites.
JOURNAL Yeast 1986;2:163-7.
PUBMED 3333305
REFERENCE 2 (bases 1 to 5181)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5181)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Yeast";
date: "1986"; volume: "2"; pages: "163-7"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5181
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 15..71
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(75..91)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(367..1167)
/codon_start=1
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
promoter complement(1168..1378)
/label=URA3 promoter
rep_origin complement(1614..2778)
/direction=LEFT
/label=2u ori
/note="yeast 2u plasmid origin of replication"
promoter 3060..3164
/label=AmpR promoter
CDS 3165..4022
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 4196..4784
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 5072..5093
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5108..5138
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5146..5162
/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)."
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