hU6 hU6(72-320) hU6 hU6(72-320) gRNA scaffold gRNA scaffold(344-419) gRNA ... gRNA scaffold(344-419) gRNA ... gRNA scaffold(344-419) cPPTCTS cPPTCTS(471-588) cPPTC... cPPTCTS(471-588) cPPTC... cPPTCTS(471-588) hPGK promoter hPGK promoter(637-1147) hPGK ... hPGK promoter(637-1147) Puro Puro(1169-1768) Puro Puro(1169-1768) SV40 late pA SV40 late pA(1906-2127) SV40 ... SV40 late pA(1906-2127) SV40 ... SV40 late pA(1906-2127) pBR322 ori pBR322 ori(2455-3043) pBR32... pBR322 ori(2455-3043) Ampicillin Ampicillin(3214-4074) Ampic... Ampicillin(3214-4074) AmpR promoter AmpR promoter(4075-4179) AmpR ... AmpR promoter(4075-4179) ...AmpR AmpR promoter(4075-4179) f1 ori f1 ori(4206-4661) f1 or... f1 ori(4206-4661) poly(A) signal poly(A) signal(4792-4840) poly(... poly(A) signal(4792-4840) ...poly( poly(A) signal(4792-4840) pause site pause site(4854-4945) pause... pause site(4854-4945) ...pause pause site(4854-4945) poly(A) signal poly(A) signal(4794-4842) poly(... poly(A) signal(4794-4842) ...poly( poly(A) signal(4794-4842) gRNA scaffold gRNA scaffold(346-421) gRNA ... gRNA scaffold(346-421) gRNA ... gRNA scaffold(346-421) pause site pause site(4856-4947) pause... pause site(4856-4947) ...pause pause site(4856-4947) AmpR promoter AmpR promoter(4077-4181) AmpR ... AmpR promoter(4077-4181) ...AmpR AmpR promoter(4077-4181) cPPT/CTS cPPT/CTS(473-590) cPPT/... cPPT/CTS(473-590) cPPT/... cPPT/CTS(473-590) f1 ori f1 ori(4208-4663) f1 or... f1 ori(4208-4663)