Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012195 | Bcr/Abl P210-pLEF | 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:
- Bcr/Abl P210-pLEF
- Antibiotic Resistance:
- Ampicillin
- Length:
- 15474 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Copy Number:
- High Copy
- Promoter:
- SV40
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- T7 or GST
- 3' Primer:
- SP6
Bcr/Abl P210-pLEF 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.
Bcr/Abl P210-pLEF vector Sequence
LOCUS V012195 15474 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V012195
VERSION V012195
KEYWORDS Bcr/Abl P210-pLEF
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 15474)
AUTHORS Kweon SM, Cho YJ, Minoo P, Groffen J, Heisterkamp N
TITLE Activity of the Bcr GTPase-activating domain is regulated through
direct protein/protein interaction with the Rho guanine nucleotide
dissociation inhibitor.
JOURNAL J Biol Chem. 2008 Feb 8;283(6):3023-30. Epub 2007 Dec 10.
PUBMED 18070886
REFERENCE 2 (bases 1 to 15474)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 15474)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
Chem."; date: "2008-02-8"; volume: "283(6)"; pages: "3023-30. Epub
2007 Dec 10"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..15474
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..211
/label="SV40 promoter"
/note="SV40 early promoter"
intron 250..822
/label="beta-globin intron"
/note="intron from rabbit beta-globin gene"
primer_bind 830..849
/label="pCAG-F"
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
promoter 877..895
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 911..1564
/label="GST"
/note="glutathione S-transferase from Schistosoma
japonicum"
CDS 1571..1588
/label="thrombin site"
/note="thrombin recognition and cleavage site"
CDS 4400..7711
/gene="ABL1"
/label="Tyrosine-protein kinase ABL1"
/note="Tyrosine-protein kinase ABL1 from Homo sapiens.
Accession#: P00519"
promoter complement(9734..9752)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
polyA_signal 9778..10001
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 10048..10476
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 10490..10819
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 10886..11677
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
polyA_signal 11854..11987
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(12024..12040)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(12024..12040)
/label="M13 Reverse"
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(12037..12059)
/label="M13/pUC Reverse"
/note="In lacZ gene"
protein_bind 12048..12064
/label="lac operator"
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(12072..12102)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(12117..12138)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(12255..12272)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(12426..13014)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(13188..14045)
/label="AmpR"
/note="beta-lactamase"
promoter complement(14046..14150)
/label="AmpR promoter"
primer_bind complement(14225..14244)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
rep_origin complement(14718..15173)
/direction=LEFT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 15314..15330
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"