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:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 52
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 775
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 946
- 1000INCOMPATIBLE 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
- Furin cut site is removed
Components:
GFP, 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