Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000588 | pWZL hygro 12S E1A | 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:
- pWZL hygro 12S E1A
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6437 bp
- Type:
- Mammalian Expression, Retroviral
- Replication origin:
- ori
- Selection Marker:
- Hygromycin
- Copy Number:
- High Copy
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- pBABE 5'
- 3' Primer:
- IRES-R
pWZL hygro 12S E1A 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.
pWZL hygro 12S E1A vector Sequence
LOCUS pWZL_hygro_12S_E 6437 bp DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pWZL hygro 12S E1A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6437)
AUTHORS Serrano M, Lin AW, McCurrach ME, Beach D, Lowe SW
TITLE Oncogenic ras provokes premature cell senescence associated with
accumulation of p53 and p16INK4a.
JOURNAL Cell. 1997 Mar 7. 88(5):593-602.
PUBMED 9054499
REFERENCE 2 (bases 1 to 6437)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6437)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 1997
Mar 7. 88(5):593-602."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6437
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 8..477
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 542..741
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 942..1358
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
CDS 1396..2124
/label=Ad5E1A
/note="Human adenovirus type 5 E1A protein gene"
misc_feature 2251..2712
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 2717..3739
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
LTR 3809..4400
/label=LTR
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(4530..4547)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4701..5289)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5452..6309)
/label=AmpR
/note="beta-lactamase"
promoter complement(6310..6414)
/label=AmpR promoter