Schematic representation of the lbk gene, positions of gRNAs, and knock-in cassette. Schematic representation of the lbk gene, positions of gRNAs, and.

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Schematic representation of the lbk gene, positions of gRNAs, and knock-in cassette. Schematic representation of the lbk gene, positions of gRNAs, and knock-in cassette. We selected two guides: one at the N-terminus between two putative start sites on the antisense strand (bottom), gRNA-lbk1, and the other overlapping the termination codon on the sense strand (top), gRNA-lbk2. The gRNA sequences are shown in black with PAM sites in red, while gray indicates the complementary sequence. The underline in gRNA-lbk2 indicates the termination codon. The knock-in cassette consisted of a 225 bp 3xFLAG-3XHA flanked by 387 and 384 bp lbk homology arms. The length of genomic lbk from the first putative ATG start codon to the TGA stop codon is 4629 bp. Nanci S. Kane et al. G3 2016;7:87-93 ©2017 by Genetics Society of America