lentiCRISPR v2-Blast (Plasmid 83480) BioVector NTCC质粒载体菌种细胞基因保藏中心

lentiCRISPR v2-Blast (Plasmid 83480) BioVector NTCC质粒载体菌种细胞基因保藏中心 Mammalian expression of Cas9 and sgRNA scaffold. lentiCas9-Blast was digested with BamHI and SacII. The resulting small fragment (encoding P2A, blasticidin and a portion of WPRE) was gel-purified. lentiCRISPR v2 was similarly digested and the larger fragment gel-purified. The two fragments combined and ligated. Vector backbone lentiCRISPR v2 (#52961) (Search Vector Database) Backbone manufacturer Feng Zhang Backbone size w/o insert (bp) 10000 Total vector size (bp) 14873 Modifications to backbone puromycin N-acetyltransferase was replaced by blasticidin S deaminase Vector type Mammalian Expression, Lentiviral, CRISPR Selectable markers Blasticidin GROWTH IN BACTERIA Bacterial Resistance(s) Ampicillin Growth Temperature 30°C Growth Strain(s) NEB Stable Growth instructions No more than 16-hour culture. Copy number High Copy GENE/INSERT Gene/Insert name Blasticidin S deaminase Mutation Replaced puromycin N-acetyltransferase on the original plasmid Promoter EF-1α CLONING INFORMATION Cloning method Restriction Enzyme 5′ cloning site BamHI (not destroyed) 3′ cloning site SacII (not destroyed) 5′ sequencing primer TGCTGAAACAAGCCGGAGAT 3′ sequencing primer AATTAGCCCTTCCAGTCCCC, WPRE_R [Supplier来源] http://www.biovector.net