Basic Vector Information
Mammalian expression of Cre recombinase from the broadly active CAG promoter/enhancer
- Vector Name:
- pCAG-Cre
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5871 bp
- Type:
- CRE/LOXP
- Replication origin:
- ori
- Source/Author:
- Dr. Connie Cepko's lab
- Copy Number:
- High copy number
- Promoter:
- chicken β-actin
- Expression Method:
- Transient
pCAG-Cre vector Vector Map
Plasmid Resuspension 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 ℃.
pCAG-Cre vector Sequence
LOCUS pCAG-Cre. 5871 bp DNA circular SYN 06-APR-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pCAG-Cre.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5871)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5871)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5871
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 4..383
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 386..661
/note="chicken beta-actin promoter"
intron 662..1670
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
CDS 1737..2765
/label=Cre
/note="site-specific recombinase"
CDS 2775..2804
/label=Myc
/note="Myc (human c-Myc proto-oncogene) epitope tag"
polyA_signal 2960..3015
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(3374..3390)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3398..3414)
/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(3422..3452)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3467..3488)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3546..3742
/label=SV40 promoter
/note="SV40 early promoter"
polyA_signal 3748..3882
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(4121..4709)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4883..5740)
/label=AmpR
/note="beta-lactamase"
promoter complement(5741..5845)
/label=AmpR promoter
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