pGEM®-3Zf(–) Vector Technical Bulletin

Literature # TB045

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 producing circular ssDNA. The pGEM®-3Zf(-) Vector contains SP6 and T7 RNA polymerase promoters flanking a region of multiple cloning sites within the alpha-peptide coding region of beta-galactosidase. Insertional inactivation of the alpha-peptide allows recombinant clones to be directly identified by color screening on indicator plates. For induction of ssDNA, bacterial cells containing pGEM®-3Zf(–) 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 SP6 Promoter Primer (Cat.# Q5011) or pUC/M13 Reverse Primer (Cat.# Q5401, Q5421).