Composite

Part:BBa_K4444010:Design

Designed by: Shreya Vallimanalan   Group: iGEM22_UPenn   (2022-10-10)


PhoCl Shed mRuby


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 52
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 769
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1654
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1288
    Illegal SapI.rc site found at 73


Design Notes

Modifications to RELEASE:

- Furin cut site is replaced with PhoCl

- Transmembrane domains, TEV cut site and ER retention motif are removed

- GFP is replaced with mRuby

Components:

mRuby, PhoCl, and transmembrane domain


Source

RELEASE: https://www.addgene.org/179642/

PhoCl: https://www.addgene.org/164037/

mRuby: Lab member's plasmids (David Gonzalez-Martinez)

References

Vlahos, Alexander E., et al. “Protease-Controlled Secretion and Display of Intercellular Signals.” Nature Communications, vol. 13, no. 1, 17 Feb. 2022, p. 912, www.nature.com/articles/s41467-022-28623-y, 10.1038/s41467-022-28623-y.

Zhang, W., Lohman, A., Zhuravlova, Y. et al. Optogenetic control with a photocleavable protein, PhoCl. Nat Methods 14, 391–394 (2017). https://doi.org/10.1038/nmeth.4222