Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V011726 | pCS2+ | 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
pCS2+ contains a powerful enhancer/promoter (simian CMV ie94). In the 5' untranslated region of the mRNA transcribed from the sCMV promoter, there is an SP6 promoter for in vitro RNA synthesis. It also has a T7 promoter in the opposite direction between the multiple cloning site and the SV40 polyadenylation site for probe synthesis. Its backbone is from pBluescriptIKS+.
pCS2+ can have high-level transient expression in multiple cell types and for in vitro transcription and translation. It is a great choice when researchers need a vector for gene expression studies in various organisms such as mammalian cells, Xenopus cells, avian cells, and zebrafish cells.
- Vector Name:
- pCS2+
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4092 bp
- Type:
- I.M.A.G.E. Consortium Plasmids
- Replication origin:
- ori
- Source/Author:
- I.M.A.G.E. Consortium
- Copy Number:
- High copy number
- Promoter:
- sCMV
- 3' Primer:
- M13 rev
- Growth Strain(s):
- JM108
- Growth Temperature:
- 37℃
pCS2+ 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
- Kalinowski J, Bachmann B, Thierbach G, Pühler A. Aspartokinase genes lysC alpha and lysC beta overlap and are adjacent to the aspartate beta-semialdehyde dehydrogenase gene asd in Corynebacterium glutamicum. Mol Gen Genet. 1990;224(3):317-324. doi:10.1007/BF00262424
pCS2+ vector Sequence
LOCUS Exported 4092 bp DNA circular SYN 03-SEP-2024
DEFINITION Vector that allows high-level transient expression in vertebrate
cells as well as in vitro transcription/translation.
ACCESSION .
VERSION .
KEYWORDS pCS2+
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4092)
AUTHORS I.M.A.G.E. Consortium
TITLE Direct Submission
REFERENCE 2 (bases 1 to 4092)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 4092)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4092
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 35..53
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
misc_feature 117..144
/label=stop codons
/note="stop codons"
/note="stop codons in all three reading frames"
polyA_signal complement(144..278)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter complement(391..409)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(430..446)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(454..470)
/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-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(478..508)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(523..544)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(832..1420)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1594..2451)
/label=AmpR
/note="beta-lactamase"
promoter complement(2452..2556)
/label=AmpR promoter
rep_origin complement(2582..3037)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3111..4092
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"