Composite

Part:BBa_K3396002

Designed by: Yuxin Liu   Group: iGEM20_NUDT_CHINA   (2020-10-26)


TRIM21-FRB

Proving that RING domain on TRIM21 can still conduce degradation on protein through recruiting ubiquitin-proteasome after replacing its PRYSPRY with DocS, we replace the DocS with FRB.

Usage and Biology

FRB is a kind of protein which can bind with FKBP induced by rapamycin. TRIM21 is an E3 ubiquitin-protein ligase which lead to degradation. To demonstrate that the Trim-Away alike method can be performed after replacing the DocS-Coh2 with FRB-FKBP interaction, we designed TRIM21-FRB. Here is the mechanism of the recombined TRIM21-FRB:

1. The GFPnano tagged with FRB combines with targeted protein.

2. TRIM21-DocS connect Coh2-GFPnano-target through the rapamycin induced DocS-Coh2 interaction.

3. The targeted protein is degraded by ubiquitin-proteasome system recruited by TRIM21.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 204
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1088
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 161
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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