Part:BBa_K4444010:Design

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