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. 2020 May 20;10(10):e3622. doi: 10.21769/BioProtoc.3622

Figure 2. Schematic representation of the nucleotide sequence flanking the target insert region.

Figure 2.

The position of the 20-nucleotide (nt) gRNA-matching site, the 3nt-protospacer adjacent motif (PAM) sequence for Cas9 binding is indicated along with the START and STOP codon. The two extra nucleotides in this particular example are included to ensure the out-of-frame CFP. The premature STOP codon is included to prevent the translation of a long-length product from the START codon on the shifted frame of the CFP coding sequence.