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A. A proto-spacer (highlighted blue and capital letters) and proto-spacer adjacent motif (PAM site; highlighted red); B. Oligonucleotides with proper overhangs (lower case letters) necessary to ligate into pPB41 ( Jiang et al., 2013 ; Burby and Simmons, 2017); C. Annealing and phosphorylation reaction to prepare a dsDNA proto-spacer for ligation into pPB41.
