Part:BBa_K4367011

FRET Protein (FP)

WORK IN PROGRESS: use pictures of FP from here: https://docs.google.com/document/d/1C8z9Y3Bf5k2NkXp4JD_NCtEUfyfqcI9ZZiLw7pQM-yQ/edit

It consists of two flourescent proteins, mNeongreen and mRuby3, that are fused together with a cleavable linker. The linker is a typical GS-linker and Tobacco Etch Virus Protease is able to cleave a target sequence within the linker. If TEV is present, it will cleave the linker and separate pair of fluorescent proteins, which will decrease the FRET phenomena. With a plate reader it will be possible to observe this.

FP acts as a reporter protein that measures the catalytic activiy of a protease. It would be relativly easy to quantify if FP is expressed as it is fluorescent, which can be used to troubleshoot if the protease is functional or not with relative ease. For example, it would be possible to check if TEVp works, and if it does, it would give information if iGal works when it is mixed with TEVp, and so on for following proteins.

Sequence and Features

Assembly Compatibility:

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

  Illegal BamHI site found at 1504
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 14
  Illegal BsaI.rc site found at 1511