Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: NAMBIN YIM
Date created: 2010-11-01 12:00:00
Date modified: 2015-05-08 01:12:11
pGEM-easy-T vector
| Types | DnaRegion |
| Roles | Plasmid_Backbone plasmid_vector |
| Sequences | BBa_K351011_sequence (Version 1) |
Description
pGEM-T-easy Vector is linearized vector with a single 3??-terminal thymidine at both ends. The T-overhangs at the insertion site greatly improve the efficiency of ligation of PCR products by preventing recircularization of the vector and providing a compatible overhang for PCR products generated by certain thermostable polymerases.pGEM-T-easy Vector is high-copy-number vector containing T7 and SP6 RNA polymerase promoters flanking a multiple cloning region within the α-peptide coding region of the enzyme β-galactosidase. Insertional inactivation of the α-peptide allows identification of recombinants by blue/white screening on indicator plates. Choice of Restriction Sites for Release of Insert: pGEM-T Easy Vector contains numerous restriction sites within the multiple cloning region. Its multiple cloning region is flanked by
recognition sites for the restriction enzymes EcoRI, BstZI and NotI, providing three single-enzyme digestions for release of the insert. Alternatively, a double-digestion may be used to release the insert from either vector.
pGEM-T Easy Vector System is supplied with 2X Rapid Ligation Buffer. Ligation reactions using this buffer may be incubated for 1 hour at room temperature. The incubation period may be extended to increase the number of colonies after transformation. Generally, an overnight incubation at 4??C produces the maximum number of transformants.