Purpose(Empty Backbone) Used for expression of sgRNA in mammalian cells
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||118055||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
Vector backbonepSpCas9(BB)-2A-Puro (PX459) V2.0 (Addgene#62988)
Vector typeMammalian Expression
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Growth Strain(s)NEB Stable
Copy numberHigh Copy
- 5′ sequencing primer AGCTGGGAGGCGACAAAAGG (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
pSpCas9(BB)-2A-Puro (PX459) V2.0 was digested with EcoRI to remove the T2A-Puro cassette, and a P2A-Blasticidin cassette was amplified by PCR, digested with EcoRI, and ligated.
The BbsI site (GAAGAC) in the blasticidin-resistance gene was mutated to GAGGAC with no change in amino acids.
gRNA inserts can be sequenced using pLKO.1 5’ primer, 5’-GACTATCATATGCTTACCGT-3’.
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:pSpCas9(BB)-2A-Blast was a gift from Ken-Ichi Takemaru (Addgene plasmid # 118055 ; http://n2t.net/addgene:118055 ; RRID:Addgene_118055)