Difference between revisions of "Part:BBa K1400003"
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<partinfo>BBa_K1400003 short</partinfo> | <partinfo>BBa_K1400003 short</partinfo> | ||
− | An improvement upon the promoter in BBa_K1164007. | + | An improvement upon the promoter in BBa_K1164007. This single input promoter has two upstream activating sequences (UAS). The third and fourth GAL4 binding site of the native pGAL1 promoter has been replaced with tetO binding sites in this version and the first and second GAL4 sites have been replaced with random sequences with identical C-G content. The Mig1 sequences that are native to the pGAL1 promoter are removed to allow transcriptional activation of the promoter in the presence of glucose in the cellular growth medium. |
+ | In cells expressing rtTA, this promoter can be used to drive transcription of a downstream gene by the addition of aTc (anhydrotetracycline). This is a weakly activating promoter. | ||
+ | It was designed to have similar expression to our modified gal promoter (BBA_K1400002). | ||
<html><img src="https://static.igem.org/mediawiki/2014/d/d3/PTRE_4_sites.png" /></html> | <html><img src="https://static.igem.org/mediawiki/2014/d/d3/PTRE_4_sites.png" /></html> |
Revision as of 22:02, 17 October 2014
pTre(4) Single input tet responsive promoter
An improvement upon the promoter in BBa_K1164007. This single input promoter has two upstream activating sequences (UAS). The third and fourth GAL4 binding site of the native pGAL1 promoter has been replaced with tetO binding sites in this version and the first and second GAL4 sites have been replaced with random sequences with identical C-G content. The Mig1 sequences that are native to the pGAL1 promoter are removed to allow transcriptional activation of the promoter in the presence of glucose in the cellular growth medium. In cells expressing rtTA, this promoter can be used to drive transcription of a downstream gene by the addition of aTc (anhydrotetracycline). This is a weakly activating promoter. 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 tetr sites and not repressing sites, so increasing aTC increases activation. Estradiol has no effect beyond auto-fluorescence. Fluorescence values were give arbitrarily by the flow cytometer software.
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