Difference between revisions of "Part:BBa K1400005"

Latest revision as of 03:25, 18 October 2014

pTRE(2) Single input tetr responsive promoter

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. 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.

Figure 1: (LEFT) The pTre promoter shows no activation upon addition of aTc (anhydrotetracycline)in cells expressing the activator, rtTA (reverse tetracycline-controlled transactivator), with the weak constitutive promoter, pMRP7. (RIGHT) In cells expressing rtTA with the strong constitutive promoter, pADH1, there is weak activation with increasing aTC concentration. Increasing concentrations of estradiol was added to compare its effects on activation with the other cognate promoters made. No increase in activation is seen with increasing estradiol due to the absence of GAL4 UAS's in this promoter.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 35
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K1400005 short</partinfo>
-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.
+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. 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.
 <html><img style="width:75%;" src="https://static.igem.org/mediawiki/2014/3/3e/Ptre%282%29.png" /></html>
-Figure 1: (LEFT) The pTre promoter shows no activation upon addition of aTc (anhydrotetracycline)in cells expressing the activator, rtTA (reverse tetracycline-controlled transactivator), with the weak constitutive promoter, pMRP7. (RIGHT) In cells expressing rtTA with the strong constitutive promoter, pADH1, there is weak activation with increasing aTC concentration. Increasing concentrations of estradiol was added to compare its effects on activation with the other cognate promoters made. No increase in activation is seen with increasing estradiol due to the absence of gal4 UAS's in this promoter.
+Figure 1: (LEFT) The pTre promoter shows no activation upon addition of aTc (anhydrotetracycline)in cells expressing the activator, rtTA (reverse tetracycline-controlled transactivator), with the weak constitutive promoter, pMRP7. (RIGHT) In cells expressing rtTA with the strong constitutive promoter, pADH1, there is weak activation with increasing aTC concentration. Increasing concentrations of estradiol was added to compare its effects on activation with the other cognate promoters made. No increase in activation is seen with increasing estradiol due to the absence of GAL4 UAS's in this promoter.
 <!-- Add more about the biology of this part here
 ===Usage and Biology===