Promega pGEM™-T and pGEM™-T Easy Vector Systems

The pGEM-T Vector is prepared by cutting Promega's pGEM-5Zf(+) Vector with EcoR V and adding a 3« terminal thymidine to both ends. These single 3«-T overhangs at the insertion site greatly improve the efficiency of ligation of a PCR product into the plasmid.

• 2X Rapid Ligation Buffer allows reactions to be completed in one hour at room temperature Blue/white screening can be used to directly identify recombinant clones, as T7 and SP6 RNA polymerase promoters flank a multiple cloning region within the α-peptide coding region for β-galactosidase, permitting insertional inactivation of the α-peptide
• f1 Origin of Replication allows the preparation of single-stranded DNA

pGEM-T Vector Systems

• Multiple cloning site is flanked by recognition sites for the restriction enzyme BstZ I Single-enzyme digestion allows release of the insert
Double digestion may also be used to release the insert from the vector

• Recognition sites for BstZ I, EcoR I, and Not I flank the insertion site Several options are available for removal of the desired insert DNA with a single restriction digestion