Difference between revisions of "Part:BBa K4444003:Design"
(→‎Design Notes)
Line 10: Line 10:
 
<u>Modifications to RELEASE:
 
<u>Modifications to RELEASE:
  
−
</u>Furin cut site is replaced with PhoCl
+
</u>- Furin cut site is replaced with PhoCl
−
Transmembrane domains, TEV cut site and ER retention motif are removed
+
- Transmembrane domains, TEV cut site and ER retention motif are removed
  
 
<u>Components:
 
<u>Components:
 
   
 
   
−
</u>(1) protein of interest (POI), (2) Phocl and (3) transmembrane domain  
+
</u> GFP, PhoCl, and transmembrane domain
−
 
+
  
 
===Source===
 
===Source===

Revision as of 07:59, 12 October 2022


PhoCl Shed 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 1660
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 60
    Illegal BsaI.rc site found at 1294


Design Notes

Modifications to RELEASE:

- Furin cut site is replaced with PhoCl - Transmembrane domains, TEV cut site and ER retention motif are removed

Components:

GFP, PhoCl, and transmembrane domain

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