Part:BBa_K1400001
PTRE(2)GX Dual input promoter. Activation at tetO binding sites, repression at gal4 sites.
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 two Tetr sites upstream of the TATA box. 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). 10bp downstream of the TATA box two Gal4 binding sites are added, which can be bound by Gal4 binding proteins or phusion activators like GEV (Gal4 binding domain, human estrogen receptor, vp16). 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.
Figure 1: Characterization of pTREgx via dual drug induction. pTREgx has 2 activating tetr sites and 2 repressing gal4 sites 10bp away from the TATA box. Thus, activation increases with aTc concentration and repression increases with estradiol concentration. The fusion protein, rtTA (reverse tetracycline-controlled transactivator), is the activator and is expressed with the weak constitutive promoter, pMRP7.
uOttawa 2015
uOttawa created a similar promoter to pTREGx in 2015 that has strong constitutive expression, but still is repressed by GEV. The part is called [http://https://parts.igem.org/Part:BBa_K1652000 pPGK1Gx], and is based on the constitutive PGK1 promoter. Find more info on the pPGK1Gx page.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 34
- 1000COMPATIBLE WITH RFC[1000]
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