Part:BBa_K3610013

NanoLuc - codon optimized for C. reinhardtii

This part is a modiified version of Part:BBa_K1159001. It has been codon optimized for the expression in C. reinhardtii.

Usage and Biology

NanoLuc is a luciferase introduced by Promega. The protein is derived from O. gracilirostris, a deep see shrimp, and has several advantages over other luciferase proteins. With only 19.1 kDa, NanoLuc is very stable and smaller than proteins like Firefly luciferase. Reactivity of NanoLuc further does not depend on ATP. NanoLuc catalizes the reaction of Furimazine to Furimamide which is accompanied by a luminescent output. This technology is very versatile and can be used for bioluminscent imaging, monitoring of protein stability, gene regulation and cell signalling and also the observation of protein-protein or protein-ligand interaction.

For our iGEM project 2020, we codon optimized this part for the expression in Chlamydomonas reinhardtii. C. reinhardtii is a green algae which is becoming more and more important in research, particularily in photosynthesis, environment sensing and genome evolution, However the organism is known to have a genome with a high GC content and a distinct codon usage, oftentimes making the expression of foreign genes difficult as this influences translation and mRNA stability.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 227
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 43
  
• 1000
  COMPATIBLE WITH RFC[1000]