Device

Part:BBa_K1078001

Designed by: Edgar Andres Ochoa Cruz   Group: iGEM13_USP-Brazil   (2013-09-12)

Strong reporter device optimized for Pichia pastoris, activated by methanol

This device is composed of three parts. The first part is a modified promoter PAOX1; it was found in a library where it showed more than 60% stronger activation when compared with the wild-type PAOX1 (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]


[edit]
Categories
//function/reporter/fluorescence
//function/sensor
Parameters
biologyPichia pastoris
device_typeSensor
outputRed Fluorescent protein