Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V018087 | pCMV-Bxb1 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pCMV-Bxb1 expresses the Bxb1 integrase under the control of the CMV promoter, catalysing the precise integration of exogenous genes into target sites within the host genome. This facilitates accurate gene delivery and the establishment of stable cell lines.
- Vector Name:
- pCMV-Bxb1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5801 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells
- Promoter:
- CMV
- 3' Primer:
- Sv40-polyA-R
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pCMV-Bxb1 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
- Anzalone AV, Gao XD, Podracky CJ, Nelson AT, Koblan LW, Raguram A, Levy JM, Mercer JAM, Liu DR. Programmable deletion, replacement, integration and inversion of large DNA sequences with twin prime editing. Nat Biotechnol. 2022 May;40(5):731-740. doi: 10.1038/s41587-021-01133-w. Epub 2021 Dec 9. PMID: 34887556; PMCID: PMC9117393.
pCMV-Bxb1 vector Sequence
LOCUS . 5801 bp DNA circular UNK 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION
VERSION
KEYWORDS .
SOURCE .
ORGANISM .
.
FEATURES Location/Qualifiers
rep_origin 2..456
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS 743..1600
/label="AmpR"
/note="β-lactamase"
protein_bind 1705..1738
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
rep_origin 1816..2404
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 2866..2896
/label="lac UV5 promoter"
/note="E. coli lac promoter with an 'up' mutation"
protein_bind 2904..2920
/label="lac operator"
/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-β-D-thiogalactopyranoside (IPTG)."
enhancer 2950..3253
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 3254..3457
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 3573..3591
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
polyA_signal 5543..5677
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"