Difference between revisions of "Part:BBa K1400005"

 
 
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<partinfo>BBa_K1400005 short</partinfo>
 
<partinfo>BBa_K1400005 short</partinfo>
  
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Two tetr sites upstream of the TATA box, replacing the 3rd and 4th gal4 sites. The first two gal4 sites were removed.  
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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.
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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.
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<html><img style="width:75%;" src="https://static.igem.org/mediawiki/2014/3/3e/Ptre%282%29.png" /></html>
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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.
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<span class='h3bb'>Sequence and Features</span>
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<span class='h3bb'><b>Sequence and Features</b></span>
 
<partinfo>BBa_K1400005 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1400005 SequenceAndFeatures</partinfo>
  

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]