Composite

Part:BBa_K3610050:Design

Designed by: Jonas Sebastian Trottmann   Group: iGEM20_UZurich   (2020-10-10)


CORE ectodomain / mCherry C-terminal for C. reinhardtii


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 93
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 148
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1071
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1917


Design Notes

Codon optimization done with SnapGene

Both mCherry and receptor sequence are codon optimized for C. reinhardtii

A flag tag was fused to the mCherry sequence for Western Blotting.

By having the self-cleaving peptide from the foot and mouth virus added to the sequence, coexpression of different proteins on the same vector is faciliated.

Source

BBa_K2136017, BBa_K3610012, BBa_K3610011, BBa_K416001, BBa_K3610019, BBa_M10019

References

Fan, Jin-Yu; Cui, Zong-Qiang; Wei, Hong-Ping; Zhang, Zhi-Ping; Zhou, Ya-Feng; Wang, Yun-Peng; Zhang, Xian-En (2008): Split mCherry as a new red bimolecular fluorescence complementation system for visualizing protein–protein interactions in living cells. In: Biochemical and Biophysical Research Communications 367 (1), S. 47–53. DOI: 10.1016/j.bbrc.2007.12.101.

Barahimipour, Rouhollah; Strenkert, Daniela; Neupert, Juliane; Schroda, Michael; Merchant, Sabeeha S.; Bock, Ralph (2015): Dissecting the contributions of GC content and codon usage to gene expression in the model alga Chlamydomonas reinhardtii. In: Plant J 84 (4), S. 704–717. DOI: 10.1111/tpj.13033.

Wang, Lei; Albert, Markus; Einig, Elias; Fürst, Ursula; Krust, Damaris; Felix, Georg (2016): The pattern-recognition receptor CORE of Solanaceae detects bacterial cold-shock protein. In: Nature plants 2, S. 16185. DOI: 10.1038/nplants.2016.185.