2752

2752 with ccdb , cut with BamH1 + PacI and reliate, transform with ccdb survival,  no colony grow on LB/amp, 

2752 cut with BamH1 + PacI,  ligate to BamH1-PacI fragment from145

colonies turn green under bluelight,

plasmi  digestion BamH1+ PacI or EcoRV + PvuII

multiple bands, could be incomplete digestion

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2752 sfGFP in pMiniT2, subcloned from 145 using BamH1-PacI, Plasmid digest with EcoRV-PvuII