Part:BBa_K1400003
pTre(4) Single input tet responsive promoter
An improvement upon the promoter in BBa_K1164007. This single input promoter has four upstream activating sequences (UAS). The four UAS sites are tetO binding sites that can bind to the tetracycline responsive activator protein, rtTA (reverse tetracycline-controlled transactivator), to induce transcription. The third and fourth GAL4 binding sites of the native pGAL1 promoter are replaced with tetO sites in this version. The Mig1 sequences that are native to the pGAL1 promoter are replaced with two tetO sites. In cells expressing rtTA, this promoter can be used to drive transcription of a downstream gene by the addition of aTc (anhydrotetracycline).
It was designed to have similar expression to our modified gal promoter (BBA_K1400002).
Figure 1: Characterization of pTRE via dual drug induction. pTRE has 4 activating tetO sites and no repressing sites, so increasing aTC increases activation. Estradiol has no effect beyond auto-fluorescence. Fluorescence values were given arbitrarily by the flow cytometer software. The fusion protein, rtTA (reverse tetracycline-controlled transactivator), is the activator and is expressed with the weak constitutive promoter, pMRP7.
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 35
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 166
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