PurposeExpression vector for sgRNAs cloned into the BsaI sites and for expression of Cas9 linked to mCherry via a T2A peptide
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||135012||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
Vector backbonepU6-(BbsI)_CBh-Cas9-T2A-mCherry (Addgene 64324), modification of pX330 (Addgene 42230)
Backbone manufacturerKuehn lab
- Total vector size (bp) 9285
Modifications to backboneSites for cloning sgRNAs changed to BsaI. Sticky ends for cloning sgRNAs are the same as the original parent vectors.
Vector typeMammalian Expression
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Growth Strain(s)NEB Stable
Copy numberHigh Copy
Gene/Insert namehumanized S. pyogenes Cas9
Specieshumanized S. pyogenes
Insert Size (bp)4272
- Promoter CBh
/ Fusion Proteins
- 3xFLAG (N terminal on insert) (N terminal on insert)
- 3x Flag (N terminal on insert)
- NLS (N terminal on insert)
- NLS (C terminal on insert)
- T2A-mCherry (C terminal on insert)
- Cloning method Unknown
- 5′ sequencing primer agggatggttggttggtggg (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- 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:pU6-(BsaI)_CBh-Cas9-T2A-mCherry was a gift from Yosef Shaul (Addgene plasmid # 135012 ; http://n2t.net/addgene:135012 ; RRID:Addgene_135012)
For your References section:Recruitment of DNA Repair MRN Complex by Intrinsically Disordered Protein Domain Fused to Cas9 Improves Efficiency of CRISPR-Mediated Genome Editing. Reuven N, Adler J, Broennimann K, Myers N, Shaul Y. Biomolecules. 2019 Oct 8;9(10). pii: biom9100584. doi: 10.3390/biom9100584. 10.3390/biom9100584 PubMed 31597252