Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V011351 | pBI-CMV1 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pBI-CMV1, this is a mammalian bidirectional promoter vector for constitutively expressing two genes at similar levels.
- Vector Name:
- pBI-CMV1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3114 bp
- Type:
- Mammalian Expression Vectors
- Replication origin:
- ori
- Source/Author:
- Clontech
- Copy Number:
- High copy number
- Promoter:
- CMV
- Growth Strain(s):
- stbl3
- Growth Temperature:
- 37℃
pBI-CMV1 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.
References
- Akhoon BA, Slathia PS, Sharma P, Gupta SK, Verma V. In silico identification of novel protective VSG antigens expressed by Trypanosoma brucei and an effort for designing a highly immunogenic DNA vaccine using IL-12 as adjuvant. Microb Pathog. 2011 Jul-Aug;51(1-2):77-87.
pBI-CMV1 vector Sequence
LOCUS pBI-CMV1. 3114 bp DNA circular SYN 01-JAN-1980
DEFINITION Mammalian bidirectional promoter vector for constitutively
expressing two genes at similar levels.
ACCESSION .
VERSION .
KEYWORDS pBI-CMV1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3114)
AUTHORS Clontech
TITLE Direct Submission
REFERENCE 2 (bases 1 to 3114)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT The gene inserted into each MCS should contain start and stop
codons.
FEATURES Location/Qualifiers
source 1..3114
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 67..370
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 371..574
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 602..663
/label=MCS 1
/note="MCS 1"
/note="multiple cloning site 1"
polyA_signal 777..858
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(1040..1628)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1802..2659)
/label=AmpR
/note="beta-lactamase"
promoter complement(2660..2764)
/label=AmpR promoter
polyA_signal complement(2797..2878)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
misc_feature 2986..3040
/label=MCS 2
/note="MCS 2"
/note="multiple cloning site 2"
promoter complement(3069..3106)
/note="minimal CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"