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