Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010739 | pCL-MFG-BRCA1 | 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:
- pCL-MFG-BRCA1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12409 bp
- Type:
- Mammalian Expression, Retroviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- LXSN primer
pCL-MFG-BRCA1 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.
pCL-MFG-BRCA1 vector Sequence
LOCUS V010739 12409 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V010739
VERSION V010739
KEYWORDS pCL-MFG-BRCA1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12409)
AUTHORS Ruffner H, Verma IM
TITLE BRCA1 is a cell cycle-regulated nuclear phosphoprotein.
JOURNAL Proc Natl Acad Sci U S A. 1997 Jul 8. 94(14):7138-43.
PUBMED 9207057
REFERENCE 2 (bases 1 to 12409)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 12409)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl
Acad Sci U S A. 1997 Jul 8. 94(14):7138-43."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12409
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..380
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 823..1180
/label="MMLV Psi"
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1245..1661
/label="gag (truncated)"
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
misc_feature 1671..2045
/label="pol region"
/note="Moloney murine leukemia virus (MMLV) pol region
containing the splice acceptor site"
CDS 2048..7636
/gene="BRCA1"
/label="Breast cancer type 1 susceptibility protein"
/note="Breast cancer type 1 susceptibility protein from
Homo sapiens. Accession#: P38398"
LTR 7889..8482
/label="LTR"
/note="long terminal repeat from Moloney murine leukemia
virus"
intron 8827..8892
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 9022..9042
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 9314..9448
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 9906..10040
/label="SV40 ori"
/note="SV40 origin of replication"
primer_bind 9968..9987
/label="SV40pro-F"
/note="SV40 promoter/origin, forward primer"
primer_bind complement(10105..10122)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(10276..10864)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(11038..11895)
/label="AmpR"
/note="beta-lactamase"
promoter complement(11896..12000)
/label="AmpR promoter"
primer_bind 12068..12086
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind 12246..12265
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"