Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010320 | pESC-TRP | 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
The pESC-TRP is a yeast episomal vector with a TRP1 marker, for galactose-regulated expression and tagging of up to two genes.
- Vector Name:
- pESC-TRP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6530 bp
- Type:
- Yeast Plasmids
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Agilent Technologies
- Copy Number:
- High copy number
- Promoter:
- GAL1,10
- Growth Strain(s):
- JM108
- Growth Temperature:
- 37℃
pESC-TRP 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.
References
- Yu KO, Jung J, Ramzi AB, Choe SH, Kim SW, Park C, Han SO. Development of a Saccharomyces cerevisiae strain for increasing the accumulation of triacylglycerol as a microbial oil feedstock for biodiesel production using glycerol as a substrate. Biotechnol Bioeng. 2013 Jan;110(1):343-7. doi: 10.1002/bit.24623.
- Honjoh K, Matsuura K, Machida T, Nishi K, Nakao M, Yano T, Miyamoto T, Iio M. Enhancement of menadione stress tolerance in yeast by accumulation of hypotaurine and taurine: co-expression of cDNA clones, from Cyprinus carpio, for cysteine dioxygenase and cysteine sulfinate decarboxylase in Saccharomyces cerevisiae. Amino Acids. 2010 Apr;38(4):1173-83.
pESC-TRP vector Sequence
LOCUS Exported 6530 bp DNA circular SYN 02-SEP-2024
DEFINITION Cloning vector pESC-TRP, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6530)
AUTHORS Lu Q.
TITLE pESC-TRP
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6530)
AUTHORS Lu Q.
TITLE Direct Submission
JOURNAL Submitted (08-MAY-1998) Technical Services, Stratagene, 11011 N.
Torrey Pines Rd., La Jolla, CA 92037, USA
REFERENCE 3 (bases 1 to 6530)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6530)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 6530)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(08-MAY-1998) Technical Services, Stratagene, 11011 N. Torrey Pines
Rd., La Jolla, CA 92037, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6530
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 191..472
/label=TRP1 promoter
CDS 473..1144
/codon_start=1
/label=TRP1
/note="phosphoribosylanthranilate isomerase, required for
tryptophan biosynthesis"
/translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
KK"
rep_origin complement(1250..1705)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
terminator complement(1784..1949)
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
CDS complement(2097..2120)
/codon_start=1
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/translation="DYKDDDDK"
promoter 2167..2831
/label=GAL1,10 promoter
/note="divergent inducible promoter, regulated by Gal4"
promoter 2842..2860
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 2878..2907
/codon_start=1
/label=Myc
/note="Myc (human c-Myc proto-oncogene) epitope tag"
/translation="EQKLISEEDL"
terminator 2937..3126
/label=CYC1 terminator
/note="transcription terminator for CYC1"
rep_origin complement(3369..3957)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4131..4988)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4989..5093)
/label=AmpR promoter
rep_origin complement(5120..6462)
/direction=LEFT
/label=2u ori
/note="yeast 2u plasmid origin of replication"