pGEM®-3Zf(+) Vector Technical Bulletin

Literature # TB086

The pGEM®-3Zf(+) Vector is a derivative of the pGEM®-3Z Vector and contains the origin of replication of the filamentous phage f1. The plasmid serves as a standard cloning vector, as a template for in vitro transcription and as a template for production of circular ssDNA. The pGEM®-3Zf(+) Vector contains SP6 and T7 RNA polymerase promoters flanking a multiple cloning region within the alpha-peptide coding region of beta-galactosidase. Insertional inactivation of the alpha-peptide allows recombinant clones to be identified directly by color screening on indicator plates. For induction of ssDNA, bacterial cells with the F´ episome (e.g., JM109, XL-1 Blue, DH5α™) containing pGEM®-3Zf(+) Vector recombinants are infected with an appropriate helper phage. The plasmid then enters the f1 replication mode, and the resulting ssDNA is exported from the cell as an encapsidated virus-like particle. The exported ssDNA can be used for mutagenesis in vitro or can be sequenced using the T7 Promoter Primer or pUC/M13 Forward Primer.