Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V006723 | pET-28b-Cas9-His | In stock (lyophilized plasmid) |
Buy one, get one free! |
Two vials of lyophilized plasmid will be delivered, each vial is about 5µg.
Basic Vector Information
- Vector Name:
- pET-28b-Cas9-His
- Antibiotic Resistance:
- Kanamycin
- Length:
- 9375 bp
- Type:
- Bacterial Expression, CRISPR
- Replication origin:
- ori
- Copy Number:
- High Copy
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- T7
pET-28b-Cas9-His 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 ℃.
pET-28b-Cas9-His vector Sequence
LOCUS pET-28b-Cas9-His 9375 bp DNA circular SYN 13-MAY-2021
DEFINITION For in vitro expression and purification of Cas9 protein.
ACCESSION .
VERSION .
KEYWORDS pET-28b-Cas9-His.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9375)
AUTHORS Gagnon JA, Valen E, Thyme SB, Huang P, Ahkmetova L, Pauli A,
Montague TG, Zimmerman S, Richter C, Schier AF
TITLE Efficient mutagenesis by Cas9 protein-mediated oligonucleotide
insertion and large-scale assessment of single-guide RNAs.
JOURNAL PLoS One. 2014 May 29;9(5):e98186. doi:
10.1371/journal.pone.0098186. eCollection 2014.
PUBMED 24873830
REFERENCE 2 (bases 1 to 9375)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 9375)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
date: "2014-05-29"; pages: "
10.1371/journal.pone.0098186. eCollection 2014"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9375
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(5..144)
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 230..252
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(249..437)
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
protein_bind complement(1212..1233)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(1249..2328)
/label=lacI
/note="lac repressor"
promoter complement(2329..2406)
/label=lacI promoter
primer_bind 2612..2631
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 2715..2733
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 2734..2758
/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)."
RBS 2773..2795
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2802..6905
/label=Cas9
/note="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
CDS 6918..6938
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6954..6971
/label=6xHis
/note="6xHis affinity tag"
terminator 7038..7085
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 7122..7577
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(7672..8484)
/label=KanR
/note="aminoglycoside phosphotransferase"
rep_origin 8606..9194
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9348..9365
/label=L4440
/note="L4440 vector, forward primer"