Part:BBa_K5005013

It is a genetically encoded NAD+ biosensor.

After verifying the feasibility of the TRACE system, we can utilize this system to evolve the key rate-limiting enzyme NAMPT involved in NAD+ biosynthesis. Before that, we require a reporting system for real-time monitoring of intracellular NAD+ concentration. Through literature search, we discovered that the team of Yuzheng Zhao from East China University of Science and Technology in Shanghai has developed the FiNad reporting system, which is specifically designed to monitor changes in NAD+ concentration both inside cells and in vivo. We obtained the ORF of FiNad from their published article, synthesized the fragment through DNA synthesis, and constructed it into the pCDH plasmid for the implementation of NAMPT-directed evolution experiments.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1200
  Illegal NgoMIV site found at 1530
  Illegal NgoMIV site found at 2034
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1495