-
PurposeMammalian expression plasmid for human codon optimized enAsCas12a (enhanced AsCas12a) encoding E174R/S542R/K548R substitutions
-
Depositing Labs
-
Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
---|---|---|---|---|---|
Plasmid | 107941 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
-
Vector backbonepCAG-CFP
-
Backbone manufacturerConnie Cepko lab (Addgene plasmid #11179)
-
Vector typeMammalian Expression
Growth in Bacteria
-
Bacterial Resistance(s)Ampicillin, 100 μg/mL
-
Growth Temperature37°C
-
Growth Strain(s)DH5alpha
-
Copy numberHigh Copy
Gene/Insert
-
Gene/Insert namehuman codon optimized enAsCas12a (E174R/S542R/K548R)
-
Alt nameAAS848
-
SpeciesAcidaminococcus sp. BV3L6
-
MutationE174R, S542R and K548R
- Promoter CAG
-
Tags
/ Fusion Proteins
- NLS (nucleoplasmin) (C terminal on insert)
- 3x HA (C terminal on insert)
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer pCAG-F (5'-GCAACGTGCTGGTTATTGTG-3')
- 3′ sequencing primer Bglob-pA-R (5'-ttttggcagagggaaaaaga-3') (Common Sequencing Primers)
Resource Information
-
Supplemental Documents
-
Articles Citing this Plasmid
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
-
For your Materials & Methods section:
pCAG-enAsCas12a(E174R/S542R/K548R)-NLS(nuc)-3xHA (AAS848) was a gift from Keith Joung & Benjamin Kleinstiver (Addgene plasmid # 107941 ; http://n2t.net/addgene:107941 ; RRID:Addgene_107941) -
For your References section:
Engineered CRISPR-Cas12a variants with increased activities and improved targeting ranges for gene, epigenetic and base editing. Kleinstiver BP, Sousa AA, Walton RT, Tak YE, Hsu JY, Clement K, Welch MM, Horng JE, Malagon-Lopez J, Scarfo I, Maus MV, Pinello L, Aryee MJ, Joung JK. Nat Biotechnol. 2019 Feb 11. pii: 10.1038/s41587-018-0011-0. doi: 10.1038/s41587-018-0011-0. 10.1038/s41587-018-0011-0 PubMed 30742127