gRNA scaffold gRNA scaffold(22-97) gRNA ... gRNA scaffold(22-97) gRNA ... gRNA scaffold(22-97) dU6-2 promoter dU6-2 promoter(116-515) dU6-2... dU6-2 promoter(116-515) dU6-2... dU6-2 promoter(116-515) 3xFLAG 3xFLAG(2993-3058) 3xFLA... 3xFLAG(2993-3058) 3xFLA... 3xFLAG(2993-3058) SV40 NLS SV40 NLS(3065-3085) SV40 ... SV40 NLS(3065-3085) SV40 ... SV40 NLS(3065-3085) Cas9 Cas9(3110-7210) Cas9 Cas9(3110-7210) nucleoplasmin NLS nucleoplasmin NLS(7211-7258) nucle... nucleoplasmin NLS(7211-7258) ...nucle nucleoplasmin NLS(7211-7258) T2A T2A(7274-7327) T2A T2A(7274-7327) T2A T2A(7274-7327) PuroR PuroR(7352-7951) PuroR PuroR(7352-7951) SV40 poly(A) signal SV40 poly(A) signal(7959-8093) SV40 ... SV40 poly(A) signal(7959-8093) ...SV40 SV40 poly(A) signal(7959-8093) ori ori(8694-9282) ori ori(8694-9282) AmpR AmpR(9453-10313) AmpR AmpR(9453-10313) AmpR promoter AmpR promoter(10314-10418) AmpR ... AmpR promoter(10314-10418) ...AmpR AmpR promoter(10314-10418) nucleoplasmin NLS nucleoplasmin NLS(7213-7260) nucle... nucleoplasmin NLS(7213-7260) ...nucle nucleoplasmin NLS(7213-7260) gRNA scaffold gRNA scaffold(24-99) gRNA ... gRNA scaffold(24-99) gRNA ... gRNA scaffold(24-99) AmpR promoter AmpR promoter(10316-10420) AmpR ... AmpR promoter(10316-10420) ...AmpR AmpR promoter(10316-10420) SV40 poly(A) signal SV40 poly(A) signal(7961-8095) SV40 ... SV40 poly(A) signal(7961-8095) ...SV40 SV40 poly(A) signal(7961-8095) ori ori(8696-9284) ori ori(8696-9284) PuroR PuroR(7354-7953) PuroR PuroR(7354-7953) AmpR AmpR(9455-10315) AmpR AmpR(9455-10315)