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URL: http://www.addgene.org/109358
Proper Citation: RRID:Addgene_109358
Bacterial Resistance: Ampicillin
Defining Citation: PMID:29524148
Vector Backbone Description: Backbone Marker:Joung lab; Backbone Size:7636; Vector Backbone:pMLM3705 (Addgene plasmid #47754).; Vector Types:Mammalian Expression, CRISPR; Bacterial Resistance:Ampicillin
Comments: The DNA fragment encoding the effector domain is synthesized by PCR and cloned between the AscI and PacI sites of pdCas9-NED to create in-frame fusion between dCas9 (D10A, H840A) and the effector domain. For experiments involving cell lines stably expressing the dCas9-effector domain fusion protein, the coding sequence of the effector domain can be excised from the pdCas9-NED-based recombinant plasmid and transferred into pHAGE EF1α dCas9-NED (Addgene plasmid #109369). Cloning the AscI-PacI fragment between the MluI and AsiSI sites of pHAGE EF1α dCas9-NED preserves the reading frame of the fusion protein (Goubert et al., 2018).
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Source: Addgene
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