Basic Vector Information
- Vector Name:
- YCplac33
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5603 bp
- Type:
- Yeast Plasmids
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Gietz RD, Sugino A.
- Copy Number:
- High copy number
- Promoter:
- URA3
- 5' Primer:
- M13 fwd
- 3' Primer:
- M13 rev
YCplac33 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 ℃.
YCplac33 vector Sequence
LOCUS 40924_49487 5603 bp DNA circular SYN 01-JAN-1980
DEFINITION Yeast centromeric plasmid with a URA3 marker.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5603)
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 5603)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5603)
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..5603
/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"
misc_feature complement(1064..2227)
/label=CEN/ARS
/note="S. cerevisiae CEN4 centromere fused to the
autonomously replicating sequence ARS1/ARS416"
CDS complement(2565..3365)
/codon_start=1
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
promoter complement(3366..3581)
/label=URA3 promoter
promoter 3697..3801
/label=AmpR promoter
CDS 3802..4659
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 4833..5421
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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