Difference between revisions of "Part:BBa K1400001"

 
 
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<partinfo>BBa_K1400001 short</partinfo>
 
<partinfo>BBa_K1400001 short</partinfo>
  
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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  
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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.
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<html><img src="https://static.igem.org/mediawiki/2014/1/17/Ptre2site.png" /></html>
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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.
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=== uOttawa 2015 ===
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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.
  
 
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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_K1400001 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1400001 SequenceAndFeatures</partinfo>
  

Latest revision as of 01:45, 19 September 2015

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


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 34
  • 1000
    COMPATIBLE WITH RFC[1000]