Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V014683 | pHRW40 | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pHRW40
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11218 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Yeast
- Selection Marker:
- Neo/G418
- Promoter:
- CMV
- Growth Temperature:
- 37℃
pHRW40 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.
pHRW40 vector Sequence
LOCUS V014683 11218 bp DNA circular SYN 01-JAN-1980 DEFINITION Exported. ACCESSION V014683 VERSION V014683 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 11218) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..11218 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 1..19 /label="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" CDS 420..2177 /gene="HO" /label="Homothallic switching endonuclease" /note="Homothallic switching endonuclease from Saccharomyces cerevisiae (strain ATCC 204508 / S288c). Accession#: P09932" CDS 2226..2723 /label="13xMyc" /note="13 tandem Myc epitope tags" terminator 2799..3046 /label="CYC1 terminator" /note="transcription terminator for CYC1" promoter complement(3117..3147) /label="lac promoter" /note="promoter for the E. coli lac operon" protein_bind complement(3162..3183) /label="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(3471..4059) /direction=LEFT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4233..5090) /label="AmpR" /note="beta-lactamase" promoter complement(5091..5195) /label="AmpR promoter" gene 5403..6759 /label="kanMX" /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" rep_origin 7102..7939 /label="ARS1" /note="S. cerevisiae autonomously replicating sequence ARS1/ARS416" misc_feature 8075..8367 /label="CEN4" /note="S. cerevisiae CEN4 centromere" CDS complement(9171..10175) /label="tTA" /note="tetracycline-controlled transactivator, comprising a fusion of the tetracycline repressor TetR with the C-terminal activation domain of herpes simplex virus VP16" promoter complement(10281..10484) /label="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" enhancer complement(10485..10864) /label="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" terminator 10977..11161 /label="ADH1 terminator" /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene"