Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Cai Hong
Date created: 2009-10-20 11:00:00
Date modified: 2015-05-08 01:11:49
pGADT7 is the AD Vector included with MATCHMAKER Two-Hybrid System 3.
| Types | DnaRegion |
| Roles | Plasmid_Backbone plasmid_vector |
| Sequences | BBa_K289009_sequence (Version 1) |
Description
The pGADT7 vector expresses proteins fused to amino acids 768???881 of the GAL4 activationdomain (AD). In yeast, fusion proteins are expressed at high levels from the constitutive ADH1
promoter (PADH1); transcription is terminated at the ADH1 transcription termination signal (TADH1). The
fusion protein is targeted to the yeast nucleus by the SV40 nuclear localization sequences that have
been added to the activation domain sequence (1). pGADT7 also contains the T7 promoter, an HA
epitope tag, and a MCS. pGADT7 replicates autonomously in both E. coli and S. cerevisiae from the
pUC and 2 m ori, respectively. The vector carries Ampr for selection in E. coli and the LEU2 nutritional
marker for selection in yeast.
Notes
Propagation in E. coli:??? Suitable host strains: DH5a, DH10 & other general purpose strains
??? Selectable marker: plasmid confers resistance to ampicillin (100 mg/ml) to E. coli hosts
??? E. coli replication origin: pUC
??? Copy number: ~500
??? Plasmid incompatibility group: pMB1/Col E1
Propagation in S. cerevisiae:
??? Suitable host strains: Y187(a), Y190(a), SFY526(a), CG1945(a), HF7c(a), or AH109(a)
??? Selectable marker: LEU2
??? S. cerevisiae origin: 2 m