Part:BBa_K1078001:Design

Strong reporter device optimized for Pichia pastoris, activated by methanol

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]

Design Notes

After synthesis, in order to build our sensor the part was introduced in pPIC9K plasmid to allow its genomic recombination into the Pichia pastoris genome, substituting the original P_AOX1 found in the yeast. The sequenced was modified to remove restriction sites, making it compatible to RFC10.

The RFP condon optimized was done using GeneScript tools at http://www.genscript.com/tools.html and the sequence from the biobrick BBa_E1010.

This part does not have a combined biobrick version with the Mxr1 because both must be inserted in different locations at the Pichia pastoris genome in order to work properly.

Source

The device was obtained by synthesis.

References

The modified pAOX1 sequence was found in Hartner FS, Ruth C, Langenegger D, Johnson SN, Hyka P, Lin-Cereghino GP, Lin-Cereghino J, Kovar K, Cregg JM, Glieder A: Promoter library designed for fine-tuned gene expression in Pichia pastoris. Nucleic Acids Res 2008, 36:e76. The RFP condon optimized was done using GeneScript tools at http://www.genscript.com/tools.html and the sequence from the biobrick BBa_E1010.