Device

Part:BBa_K5114228:Design

Designed by: James Marshall, Douglas Lin, Kalp Poladia, Vishwaa Kannan, Daniel Jiang, Leon Guo   Group: iGEM24_GCM-KY   (2024-09-23)


Expression device for BBa_K5114227 (hlFAB-GFP)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

When we asked the original creators for the sequence of the protein, they gave us one that was slightly different from the one that we reconstructed from their paper. Additionally, they recommended that we use a stronger promoter to express the protein.


Source

The GFP-FAB conjugation sequence was originally characterized by Mann and Berger in 2013 and used to test for absorbance upon titration of PFOA. FAB is a human liver fatty acid binding protein, and is found within the cytoplasm of human liver cells and mediates the intercellular transport of various fatty acids.

References

  Mann, M. M., & Berger, B. W. (2023, September 13). A genetically-encoded biosensor for direct detection of perfluorooctanoic acid. Nature News. https://www.nature.com/articles/s41598-023-41953-1 

  Smathers, R. L., & Petersen, D. R. (2011, March 1). The human fatty acid-binding protein family: Evolutionary divergences and functions - human genomics. BioMed Central. https://humgenomics.biomedcentral.com/articles/10.1186/1479-7364-5-3-170#:~:text=Fatty%20acid%2Dbinding%20proteins%20