Fig. 6

Overview of CRISPR/Cas9-BB3 plasmids assembled using GoldenPiCS. Transcription units for sgRNA (fusion of crRNA and tracrRNA [42]) and hcas9 (with SV40 nuclear localization sequence at its C-terminus) with various promoters and transcription terminators were assembled in BB3cK_AC (CEN/ARS locus, KanMX selection marker, fusion sites FsA-FsC). The plasmids were successfully used to generate InDel mutations in different genomic loci and eGFP in P. pastoris. The effects of the tested constructs are summarized in the table below the vector scheme. Modules which worked most efficiently are indicated in bold in the vector scheme