Part:BBa_K1164008

pTRELX 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 The four gal4 binding domains present in the native Gal1 promoter have been replaced by four tetO sites.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. In addition, a lacO operator has been introduced downstream of the TATA box, allowing for repression of gene expression by lacI. Upon expression from the promoter, yEGFP will be produced.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 465
  Illegal BamHI site found at 553
  
• 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 1202
  

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The above data was collected from the following yeast strain:
ade2::Kan-rtACT1-pGalLX-rtTA-mCherry-VP16 ade4::Nat-rtACT1-pTRE-LacI-BFP
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.
In the above construct, PTRE (driving BFP) is activated by rtTA , which is driven by pGallx and requires the presence of aTc. The levels of fluorescence then stabilize and achieve a steady level.