Coding

Part:BBa_K3610015

Designed by: Jonas Sebastian Trottmann   Group: iGEM20_UZurich   (2020-09-13)


LargeBit NanoLuc - codon optimized for C. reinhardtii

This part is a modified version of Part:BBa_K1761005. It has been codon optimized for the expression in C. reinhardtii.

Usage and Biology

NanoLuc is a small, bright and stable luciferase from O. gracilirostris and has been proven to be an efficient reporter protein since its introduction by Promega. One way to use the NanoLuc luciferase is in a protein-fragement complementation assay (PCA) to visualize protein-protein or protein-ligand interaction. The protein-protein interaction is observed by fusing spli-reporter proteins, like the LargeBit and the SmallBit from NanoLuc, to the proteins of interest. Interaction of the two proteins will associate the reporter fragments and lead to a reconstitution of the full length and fully functional NanoLuc protein.

To enhance translation, we codon optimized this part for the expression in C. reinhardtii as codon bias has shown to influence mRNA stability and translational efficiacy. C. reinhardtii has a strong codon bias and additionally a high GC content, oftentimes making efficient expression of foreign sequences in C. reinahrdtii very difficult.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None