Coding

Part:BBa_K4830027:Design

Designed by: Ong Jie Hao   Group: iGEM23_NTU-Singapore   (2023-10-11)


mCherry_C-terminus stop codon_Clover


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 352
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 352
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 352
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 352
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Given that the stop codon is installed at the C-terminus of the mCherry coding sequence, the mCherry protein is still expressed while the GFP is not expressed. This is particularly useful in assays that still require the presence of mCherry for detection.


Source

The mCherry fluorescent protein is derived from Discosoma sp. It has an excitation maxima at 587nm, and emission maxima of 610nm.

The Clover fluorescent protein (GFP) is derived from Aequorea victoria. It has an excitation maxima of 498nm, and emission maxima of 509nm.

References

Certo, M. T., Ryu, B. Y., Annis, J. E., Garibov, M., Jarjour, J., Rawlings, D. J., & Scharenberg, A. M. (2011). Tracking genome engineering outcome at individual DNA breakpoints. Nature Methods, 8(8), 671–676.