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Targeted Large-Scale Deletion of Bacterial Genomes Using CRISPR-Nickases.
Standage-Beier K, Zhang Q, Wang X
ACS Synth Biol. 2015 Nov 20;4(11):1217-25. doi: 10.1021/acssynbio.5b00132. Epub 2015 Oct 25.
PubMed Article

Plasmids from Article

ID Plasmid Purpose  
74492pSG4K5For cloning and constitutive expression of sgRNAs in E.coli. New sgRNAs are cloned into SapI sites and multiplex sgRNAs are cloned into BsaI sites. Kanamycin resistant with pSC101 origin of rep.
74495pCas9(D10A)Nicking Cas9 derived from pWT Cas9 (Addgene #44250). Ampicillin resistance marker, the tetracycline repressor (TetR), a ColE1 origin of replication and Streptococcus pyogenes Cas9 D10A