|
Download
protocol
188kb
pdf? |
|
Abstract for pGEM®-5ZF(+) Vector
The pGEM®-5Zf(+) Vector is a derivative of the pGEM®-3Zf(+)
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 can be used for the production of circular ssDNA. pGEM®-5Zf(+)
contains T7 and SP6 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. The multiple cloning region
contains unique restriction sites for ApaI,
AatII, SphI,
NcoI, SacII,
EcoRV, SpeI,
NotI, PstI,
SalI, NdeI,
SacI, BstXI and
NsiI. This arrangement is designed specifically for generating
unidirectional deletions with Promega’s Erase-a-Base® System. The
polylinker contains restriction enzyme sites that produce 5´ overhangs or blunt
ends (sensitive to Exonuclease III), flanked on both sides by blocks of
restriction sites that generate 3´ overhangs (resistant to Exonuclease III).
|
