Composite

Part:BBa_K4444002:Design

Designed by: Cristina Perez, Adiva Daniar, June Ahn, Wangari Mbuthia, Shreya Vallimanalan   Group: iGEM22_UPenn   (2022-10-10)


PhoCl Secrete GFP


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 775
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 946
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 60
    Illegal BsaI.rc site found at 1468


Design Notes

Modifications to RELEASE:

- TEV cut site is replaced with PhoCl

Components:

GFP, furin cut site, tri-transmembrane domain, PhoCl, and ER retention motif

Source

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

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

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