Generator

Part:BBa_K3580102:Design

Designed by: Yuya Otsuki   Group: iGEM20_Waseda   (2020-10-23)
Revision as of 04:20, 24 October 2020 by Kanta (Talk | contribs)


Ptrc-trGPPS-SS(Sabinene synthase)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1980
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1980
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2366
    Illegal XhoI site found at 4022
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1980
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1980
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part was designed based on the Ptrc-trGPPS-LS(BBa_K3580101)used in the 2013 paper by Alonso-Gutierrez, Jorge et al. The introduction of the one-amino acid mutation reported in the2015 paper by Srividya, Narayanan et al.The one-amino acid mutation was introduced into the limonene synthase of Mentha spicatato accommodate the N345A mutation, as reported in a 2015 paper by Srividya and Narayanan et al.Actually, limonene synthase used in our project was an E. colicodon-optimized version of a truncated sequence, so a mutation was introduced to change asparagine at position 289 to alanine.


Source

Improved for BBa_K3052001so that it can synthesize sabinene and be used in combination with an addgenene plasmid (pBbA5c-MevT-MBI) encoding a series of enzymes in the mevalonate pathway. The products of the mevalonate pathway (IPP and DMAPP) can also be synthesized by the non-mevalonate pathway. This part has improved modularity in the sense that it can use both the mevalonate and non-mevalonate pathways for upstream metabolic pathways.

References