Generator

Part:BBa_K3580103:Design

Designed by: Yuya Otsuki   Group: iGEM20_Waseda   (2020-10-26)


A series of enzymes of the mevalonate pathway


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1532
    Illegal XbaI site found at 5511
    Illegal XbaI site found at 6620
    Illegal XbaI site found at 8076
    Illegal XbaI site found at 8969
    Illegal SpeI site found at 10120
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1532
    Illegal SpeI site found at 10120
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1532
    Illegal BglII site found at 1541
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1532
    Illegal XbaI site found at 5511
    Illegal XbaI site found at 6620
    Illegal XbaI site found at 8076
    Illegal XbaI site found at 8969
    Illegal SpeI site found at 10120
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1532
    Illegal XbaI site found at 5511
    Illegal XbaI site found at 6620
    Illegal XbaI site found at 8076
    Illegal XbaI site found at 8969
    Illegal SpeI site found at 10120
    Illegal NgoMIV site found at 1418
    Illegal NgoMIV site found at 5656
    Illegal AgeI site found at 3374
    Illegal AgeI site found at 6852
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part is a converted version of pBbA5c-MevT-MBI created and used in a paper reported by Alonso-Gutierrez, Jorge et al. in 2013 to make Biobrick RFC1000 compatible.We obtained expression system for those seven genes from addgene and have converted this into Biobrick RFC 1000 format by synonymous replacement


Source

Improved for BBa_K3052010so that it can be used in combination with BBa_K3580101 (Ptrc-trGPPS-LS) and BBa_K3580102 (Ptrc-trGPPS-SS).The products of the mevalonate pathway (IPP and DMAPP) can also be synthesized by the non-mevalonate pathway. This part is more modular, for example in the sense that it can be replaced by enzymes in the non-mevalonate pathway.

References

(1) Alonso-Gutierrez, J. et al. (2013). Metabolic engineering of Escherichia coli for limonene and perillyl alcohol production. Metabolic engineering, 19, 33-41.

(2) Dudley, Q. M. et al. (2019). Cell-free biosynthesis of limonene using enzyme-enriched Escherichia coli lysates. Synthetic Biology, 4(1), ysz003.

(3) Mass Spectrometry Data Center, William E. Wallace, "Mass Spectra" in NIST Chemistry WebBook, NIST Standard Reference Database Number 69, Eds. P.J. Linstrom and W.G. Mallard, National Institute of Standards and Technology, Gaithersburg MD, 20899, https://doi.org/10.18434/T4D303, (retrieved October 22, 2020).