Part:BBa_K1164007

pTRE driving yEGFP with tPGK1 terminator

This device consists of a modified version of the native Gal1 promoter found in S.cerevisiae driving production of yeast codon optimized GFP. The four gal4 binding domains present in the native Gal1 promoter have been replaced by four tetO sites. This promoter may be activated by regulatory activators that are able to bind to tetO. Upon activation, yEGFP will be produced.

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 81
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1110
  

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The above data was collected from the following yeast strain:
ade2::Kan-rtACT1-pGal1-rtTA-mCherry-VP16 ade4::Nat-rtACT1-pTRE-LacI-BFP In the above construct, PTRE (driving BFP) is activated by rtTA , which is driven by pGal and requires the presence of aTc. The levels of fluorescence then stabilize and achieve a steady level. The levels of fluorescence then stabilize and achieve a steady level. This graphic shows that as the aTc concentration increases, PTRE is being increasingly activated, resulting in greater BFP transcription. This is expressed as the increase of fluorescence in the graphic above.

Reference

1. Ellis T, Wang X, Collins JJ. Diversity-based, model-guided construction of synthetic gene networks with predicted functions. Nat Biotechnol. 2009;27:46