Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Dylan Siriwardena
Date created: 2014-10-05 11:00:00
Date modified: 2015-05-08 01:10:16
pGALtx Dual input promoter. Activation at gal4 binding sites, repression at tetO sites.
| Types | DnaRegion |
| Roles | Regulatory promoter |
| Sequences | BBa_K1400004_sequence (Version 1) |
Description
This is a dual input promoter that can be activated and repressed by two different activating proteins (Tet and Gal4). Is based off the Gal promoter in Saccharomyces cerevisiae, which is a relatively strong promoter. It has four gal4 sites upstream of the TATA box, which can be bound by Gal4 binding proteins or phusion activators like GEV (Gal4 binding domain, human estrogen receptor, vp16). 10bp downstream of the TATA box two tetr binding sites are added. These can be bound by the tet repressor, or by activating variants using the Tetr binding domain like rtTA (tet binding domain, vp16 activating domain).The close proximity of these sites to the TATA box cause any binding protein (activating or repressing) to repress expression. The close proximity of these sites all seem to affect transcription rates of this promoter, as it has significantly less expression than the native Gal promoter. Thus this promoter can be used for complex control of expressionNotes
The major design consideration was the proximity of the repressing sites to the TATA box. 10bp seemed to cause strong repression while still not interfering too much with transcription when no proteins are bound.Source
This part was generated by modifying the native Gal promoter from Saccharomyces cerevisiae.| Sequence Annotation | Location | Component / Role(s) |
| GAL4 binding site GAL4 binding site GAL4 binding site GAL4 binding site TATA box tetO tetO | 7,23 26,42 44,60 108,124 335,340 351,369 372,390 | feature/operator operator feature/operator operator operator feature/operator sequence_feature feature/misc feature/misc sequence_feature feature/operator operator operator feature/operator |