Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Stephen A Rettie
Date created: 2014-10-08 11:00:00
Date modified: 2015-05-08 01:10:18
Gal4-Degron-Vp16
| Types | DnaRegion |
| Roles | CDS Coding |
| Sequences | BBa_K1408002_sequence (Version 1) |
Description
Gal4 protein binds to a Gal1 promoter while VP16 recruits transcription machinery promoting transcription to anything under the Gal1 promoter. The two must be co-localized to work as a transcriptional activator.The degron protein domain when fused to a protein acts as a source of instability leading to degradation by the cell via ubiquitination.
This system can be used to compare the relative stabilities of proteins. A sequence for a protein inserted between the degron and VP16 will produce a fusion protein which acts as a transcriptional activator for anything under a Gal1 promoter. If the protein inserted is unstable it will be degraded by the cell and the Gal4-VP16 transcriptional activator will no longer function. If a quantifiable marker protein such as GFP is under Gal1 then the level of GFP output of the cell is related to the stability of the inserted protein.
Notes
Xbe1, EcoR1 and Spe1 restriction sites were present in the original sequence obtained from the Fields Lab. These are illegal restriction sites according to iGEM rules. The submitted part was synthesized with synonymous substitutions (silent mutations) amplified with primers containing the biobrick prefix and suffix and then digested and ligated into the iGEM vector.Source
The degron is a Protein domain Saccharomyces cerevisiae MATa2 (a=alpha).Gal4 is a DNA binding fragment from Saccharomyces cerevisiae. VP16 is a transcriptional activator from herpes simplex virus.
These parts were obtained from the Fields Lab at the University of Washington.