Part:BBa_K1078001

Strong reporter device optimized for Pichia pastoris, activated by methanol

This device is composed of three parts. The first part is a modified promoter P_AOX1; it was found in a library where it showed more than 60% stronger activation when compared with the wild-type P_AOX1 (found as BBa_K431007 in the registry of parts), as reported by Hartner FS et al., in Promoter library designed for fine-tuned gene expression inPichia pastoris. Nucleic Acids Res 2008, 36:e76. The second part is a kozak sequence for P. pastoris, and the third is a red fluorescent protein condon optimized for expression in P. pastoris.

This device was created to express a reporter gene, in this case RFP, in the presence of methanol. Its expression is inactivated by ethanol. When combined with the modified Mxr1 (BBa_K1078000), it is not inactivated by ethanol. This is useful for our biosensor design, which aims to detect levels of methanol above 2% in common alcoholic drinks (normally containing 10 to 60% ethanol). This will allow government to make a preliminary high-throughput screening of ethanol drinks tainted will methanol.

Sequence and Features

Assembly Compatibility:

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

  Illegal BglII site found at 1
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]