Fig. 6.

pKIR1.1 construction. (A) For pKIR1.1, U6.26p::2 × AarI:sgRNA was inserted into the SbfI site of pKIR1.0. (B) Flowchart of pKIR1.1-based vector construction. The upper sequence is an enlarged view of the boundary (yellow) between the U6.26 promoter (pale green) and the sgRNA scaffold (orange). This boundary contains an inverted repeat of AarI recognition sites, and AarI digestion generates four-base overhangs. A hybridized primer with overhangs can be inserted into this site as shown. G with a red triangle is the first base for RNA transcription from the U6.26 promoter. G + (N)₁₉ indicates the target sequence.