Catalog  |  Cart  |  Log In


Technical Resources

Protocols

Abstract for pGEM®-3Zf(+) Vector

Part# TB086
Printed in USA. Revised 5/09.
Instructions for Use of Product P2271.

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 the 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 directly identified 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.


PDF icon
Complete
protocol

166kb