PurposeBacterial expression plasmid for S. aureus Cas9 & sgRNA (need to clone in spacer into BsaI sites): T7-humanSaCas9-NLS-3xFLAG-T7-BsaIcassette-Sa-sgRNA
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||65770||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Backbone manufacturerEMD Millipore
Vector typeBacterial Expression, CRISPR
Growth in Bacteria
Growth Strain(s)XL1 Blue
Copy numberLow Copy
Gene/Insert namemammalian codon-optimized Staphylococcus aureus Cas9-NLS-3xFlag, and SaCas9 gRNA
- Promoter T7 (x2)
/ Fusion Proteins
- NLS (C terminal on insert)
- 3x FLAG (C terminal on insert)
- Cloning method Gibson Cloning
- 5′ sequencing primer ACYC-DuetUP1
- 3′ sequencing primer oBK390 (5'-GCAAATTCGACCCGGTCGTCG-3') (Common Sequencing Primers)
Use BsaI sites to insert sgRNA spacer sequence into the vector.
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:BPK2101 was a gift from Keith Joung (Addgene plasmid # 65770 ; http://n2t.net/addgene:65770 ; RRID:Addgene_65770)
For your References section:Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Kleinstiver BP, Prew MS, Tsai SQ, Topkar VV, Nguyen NT, Zheng Z, Gonzales AP, Li Z, Peterson RT, Yeh JJ, Aryee MJ, Joung JK. Nature. 2015 Jun 22. doi: 10.1038/nature14592. 10.1038/nature14592 PubMed 26098369