pgbkt7 pGADT7
pGADT7 AD Vector Information
PT3249-5
Cat. Nos. 630442
630489 630491
800.662.2566+1.650.919.7300+33.(0)1.3904.6880+81.(0)77.54
3.6116akara Bio CompanyTerra Bella Ave. Ampr
T7 Sequencing Primer
T7 Promoter
1858
a.a.881
START
in vitro
19151969
CAT ATG GCC ATG GAG GCC AGT GAA TTC CAC CCG
1
GGT GGG CAT CGA TAC GGG ATC CAT
XmaI
2026
2081
pGADT7 AD Vector Map and Multiple Cloning Site (MCS). Description
pGADT7 AD is a yeast expression vector that is designed to express a protein of interest fused to a GAL4 activation domain (AD; amino acids 768–881). Transcription of the
GAL4 AD fusion is driven by the constitutively active ADH1 promoter (PTADH1), and is terminated at the ADH1 transcription termination signal (ADH1). The GAL4 AD fusion contains an N-terminal SV40 nuclear localization signal (SV40 NLS; 1) that targets the protein to the yeast nucleus, and a hemagglutinin epitope tag (HA Tag), located between the GAL4 AD and the protein of interest, that allows the protein to be easily detected with HA-tag antibodies.
The T7 promoter (PT7), located just upstream of the HA tag sequence, allows in vitro tran-scription and translation of the HA-tagged protein of interest (without the GAL4 AD and the SV40 NLS). pGADT7 AD replicates autonomously
2
in both E. coli and S. cerevisiae from the pUC and 2
3
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