Part:BBa_K3064014

hsaGlucagon->GGGGS->hsaIgG-Fc->P2A->HA->hsaTrim21

This is a functional unit of the HepG2, which expresses fusioned hsaGlucagon-GGGGS-hsaIgG-Fc and HA-hsaTrim21 by automatic cleavage of P2A.

Usage and Biology

Our composite part BBa_K3064014 play a necessary effect in our whole RECEPTOR KILLER system. This part is designed to recognised and bind with target protein,and then recruit the proteasome to deplete the target protein through the ubiquitin-proteasome pathway.
In this part, hsaGlucagon acts as a bait,which can bind with GCGR, and is linked together with hsalgG-Fc by the designed 4xGS linker to form a recombinate GFP antibody.
Trim21, the E3 ubiquitin ligase, plays the most important role in the degradation system. The C-terminal B30.2 domain on trim21 offers a site for the conservative Fc region of human lgG 1,2 and 4 to bind with[1]. When Glucagon is linked to the hsalgG1-Fc, and after the antibody-antigen interaction, a ternary complex is build up. The trim21 then functions as a E3 ubiquitin ligase and proceeds the complex to be depleted through the ubiquitin-proteasome pathway. Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1830
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 2499
  Illegal BamHI site found at 3037
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1787
  Illegal AgeI site found at 632
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1142
  Illegal BsaI site found at 2877
  Illegal SapI.rc site found at 1490