Composite

Part:BBa_K2971004:Design

Designed by: Simen Hermansen   Group: iGEM19_UiOslo_Norway   (2019-10-16)
Revision as of 21:45, 21 October 2019 by Simen (Talk | contribs) (→‎Design Notes)


crtEBI under pBad promotor


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
    Illegal NheI site found at 4181
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2854
    Illegal BamHI site found at 1144
    Illegal BamHI site found at 2093
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1295
    Illegal SapI site found at 961


Design Notes

Codon optimized for expression in Escherichia coli and iGEM registry compatibility.

The slightly unusual and simplified design, without an individual RBS for each gene, was to create a system where we could, with relative ease, produce several composite parts that produce various carotenoids. By combining crtEB with a phytoene desaturase of our choice, we could control what carotenoid that would be produced. When combined with CrtI from D. radiodurans lycopene was produced in cultures expressing the composite part. If, for example, it was combined with CrtI from Rhodobacter capsulatus neurosporene would be produced instead. Xu et al., 2018 showed that this design was viable for producing plasmids intended for lycopene production.


References

Xu, X., Tian, L., Xu, J., Xie, C., Jiang, L., & Huang, H. (2018). Analysis and expression of the carotenoid biosynthesis genes from Deinococcus wulumuqiensis R12 in engineered Escherichia coli. AMB Express, 8(1), 94-94. doi:10.1186/s13568-018-0624-1

Source

deinococcus radiodurans

References