Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V017973 | pCAG-Cre-Myc | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pCAG-Cre-Myc is an expression vector that simultaneously expresses Cre recombinase and a Myc tag under the control of the CAG strong promoter. It is employed for studies involving conditional gene knockout and protein expression tracking.
- Vector Name:
- pCAG-Cre-Myc
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5877 bp
- Type:
- Gene-editing vector
- Replication origin:
- ori
- Host:
- Mammalian cells
- Promoter:
- CAG
- 3' Primer:
- pcaggs-R
- Growth Strain(s):
- Top10
- Growth Temperature:
- 37℃
pCAG-Cre-Myc 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
- Nectow AR, Moya MV, Ekstrand MI, Mousa A, McGuire KL, Sferrazza CE, Field BC, Rabinowitz GS, Sawicka K, Liang Y, Friedman JM, Heintz N, Schmidt EF. Rapid Molecular Profiling of Defined Cell Types Using Viral TRAP. Cell Rep. 2017 Apr 18;19(3):655-667. doi: 10.1016/j.celrep.2017.03.048. PMID: 28423326; PMCID: PMC5476221.
pCAG-Cre-Myc vector Sequence
LOCUS . 5877 bp DNA circular UNK 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION
VERSION
KEYWORDS .
SOURCE .
ORGANISM .
.
FEATURES Location/Qualifiers
enhancer 5..384
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 386..661
/label="chicken β-actin promoter"
intron 662..1674
/label="chimeric intron"
/note="chimera between introns from chicken β-actin and
rabbit β-globin"
CDS 1741..2769
/label="Cre"
/note="site-specific recombinase"
CDS 2779..2808
/label="Myc"
/note="Myc (human c-Myc proto-oncogene) epitope tag"
polyA_signal 2964..3019
/label="β-globin poly(A) signal"
/note="rabbit β-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(3380..3396)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3404..3420)
/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)."
promoter complement(3428..3458)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(3473..3494)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3552..3748
/label="SV40 promoter"
/note="SV40 early promoter"
polyA_signal 3754..3888
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(4127..4715)
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4889..5746)
/label="AmpR"
/note="β-lactamase"
promoter complement(5747..5851)
/label="AmpR promoter"