Basic Vector Information
- Vector Name:
- YCp50-poly
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7060 bp
- Type:
- Yeast Plasmids
- Replication origin:
- ori
- Host:
- Yeast
- Copy Number:
- High copy number
- Promoter:
- URA3
- 3' Primer:
- M13 rev
YCp50-poly 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 ℃.
YCp50-poly vector Sequence
LOCUS 40924_49442 7060 bp DNA circular SYN 01-JAN-1980
DEFINITION Derivative of yeast centromeric vector YCp50 containing the pUC19
polylinker.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7060)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 7060)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT YCp50-poly was constructed in two steps: (1) YCp50 was cleaved with
XhoI and Asp718, blunted and ligated. (2) This construct was then
cleaved with EcoRI and SalI, and the EcoRI-PvuII fragment of pUC19
was inserted.
FEATURES Location/Qualifiers
source 1..7060
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 1..57
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(70..86)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(94..110)
/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)."
promoter complement(118..148)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(163..184)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1021..1241
/label=URA3 promoter
CDS 1242..2042
/codon_start=1
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
misc_feature complement(3390..3682)
/label=CEN4
/note="S. cerevisiae CEN4 centromere"
rep_origin complement(4315..4768)
/direction=LEFT
/label=ARS1
/note="S. cerevisiae autonomously replicating sequence
ARS1"
misc_feature 4910..5050
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(5236..5824)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5998..6855)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6856..6960)
/label=AmpR promoter
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