Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010187 | YEp353 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- YEp353
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7944 bp
- Type:
- Yeast Plasmids
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Myers AM, Tzagoloff A, Kinney DM, Lusty CJ.
- Copy Number:
- High copy number
- Promoter:
- URA3
- Growth Strain(s):
- Top10
- Growth Temperature:
- 37℃
YEp353 vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
YEp353 vector Sequence
LOCUS YEp353. 7944 bp DNA circular SYN 01-JAN-1980
DEFINITION Yeast episomal vector with a URA3 marker for construction of lacZ
fusions. For other reading frames, use YEp354 or YEp355.
ACCESSION .
VERSION .
KEYWORDS YEp353
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7944)
AUTHORS Myers AM, Tzagoloff A, Kinney DM, Lusty CJ.
TITLE Yeast shuttle and integrative vectors with multiple cloning sites
suitable for construction of lacZ fusions.
JOURNAL Gene 1986;45:299-310.
PUBMED 3026915
REFERENCE 2 (bases 1 to 7944)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7944)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene";
date: "1986"; volume: "45"; pages: "299-310"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT The GenBank sequence was edited at base 3044 to eliminate a second
EcoRI site.
FEATURES Location/Qualifiers
source 1..7944
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 1..36
/label=MCS
/note="MCS"
/note="multiple cloning site"
CDS 45..3092
/label=lacZ
/note="beta-galactosidase"
CDS complement(3293..4093)
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
promoter complement(4094..4304)
/label=URA3 promoter
rep_origin complement(4541..5705)
/direction=LEFT
/label=2u ori
/note="yeast 2u plasmid origin of replication"
promoter 5987..6091
/label=AmpR promoter
CDS 6092..6949
/label=AmpR
/note="beta-lactamase"
rep_origin 7123..7711
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"