Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010322 | pESC-HIS | 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-HIS plasmid is an episomal vector used in yeast. It carries a HIS3 selectable marker, enabling galactose-inducible gene expression and tandem tagging of up to two genes of interest.
- Vector Name:
- pESC-HIS
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6706 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-HIS vector 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
- Wang C, Poborsky M, Crocoll C, Nødvig CS, Mortensen UH, Halkier BA. Comparison of Genome and Plasmid-Based Engineering of Multigene Benzylglucosinolate Pathway in Saccharomyces cerevisiae. Appl Environ Microbiol. 2022 Nov 22;88(22):e0097822. doi: 10.1128/aem.00978-22. Epub 2022 Nov 3. PMID: 36326240; PMCID: PMC9680641.
pESC-HIS vector Sequence
LOCUS 40924_17601 6706 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pESC-HIS, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6706)
AUTHORS Lu Q.
TITLE pESC-HIS
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6706)
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 6706)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6706)
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"
FEATURES Location/Qualifiers
source 1..6706
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 317..503
/label=HIS3 promoter
CDS 504..1160
/codon_start=1
/label=HIS3
/note="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
rep_origin complement(1423..1878)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
terminator complement(1961..2126)
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
CDS complement(2274..2297)
/codon_start=1
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/translation="DYKDDDDK"
promoter 2344..3008
/label=GAL1,10 promoter
/note="divergent inducible promoter, regulated by Gal4"
promoter 3019..3037
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 3055..3084
/codon_start=1
/label=Myc
/note="Myc (human c-Myc proto-oncogene) epitope tag"
/translation="EQKLISEEDL"
terminator 3114..3303
/label=CYC1 terminator
/note="transcription terminator for CYC1"
rep_origin complement(3546..4134)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4308..5165)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5166..5270)
/label=AmpR promoter
rep_origin complement(5297..6639)
/direction=LEFT
/label=2u ori
/note="yeast 2u plasmid origin of replication"