BBa_K1400004

BBa_K1400004 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K1400004
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 expression

Notes

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
igem#experience
None
 
igem#sampleStatus
It's complicated
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K1400004/1