Fig. 1

The procedure of genome editing with one plasmid and one transformation. The first step is to assemble the editing plasmid. For the second step, the plasmid is transformed into E. coli, followed by plating on LB+ Kan+ 1% (w/v) glucose plate, and grew at 30 °C. For the third step, the resulting strain is grown in LB+ Kan for 2 h before induction with 2 g/L l-arabinose for Cas9 and gRNA expression. After culturing of 6 h, the culture is plated on LB+ Kan+ l-arabinose agar plates. Last step, the temperature sensitive editing plasmid is cured by growing the edited strains at 37 °C overnight