Difference between revisions of "Part:BBa K3396009"

Revision as of 12:10, 25 October 2020

GFPnano-DocS-P2A-Trim21-Coh2

CMV->Kozak->Truncated TRIM21->DocS->P2A->Coh2->GFPnano->BGH-pA

This part is designed to prove that our Trim21 truncation strategy works as we expected. Since the original Trim-away system utilizes the Trim21 PRYSPRY domain to interact with the antibody IgG, we herein changed such interaction pair into other constitutively dimerization pairs to validate our design.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1790
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 1747
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 721

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K3396009 short</partinfo>
-Usage and Biology
+CMV->Kozak->Truncated TRIM21->DocS->P2A->Coh2->GFPnano->BGH-pA
-The PRYSPRY-lgG Fc interaction of natural Trim21-antibody complex is replaced with DocS-Coh2 to demonstrate that the Trim-Away alike method could still work. DocS and Coh2 are a pair of proteins that has been widely used in synthetic biology to test split protein approach due to their high constitutively dimerization affinity. Theoretically, the dimerization of DocS and Coh2 will bring the truncated Trim21 protein and the target protein EGFP into proximity and form a s stable trimer. Truncated Trim21 would then function as the E3 ligase to ubiquitylate EGFP and mediate its degradation.
-We expect such system to work in a similar manner as the previous GFP predator as we reported in iGEM 2018.
+This part is designed to prove that our Trim21 truncation strategy works as we expected. Since the original Trim-away system utilizes the Trim21 PRYSPRY domain to interact with the antibody IgG, we herein changed such interaction pair into other constitutively dimerization pairs to validate our design.
-https://2020.igem.org/wiki/images/d/d0/T--NUDT_CHINA--GFPnano-DocS-P2A-Coh2-Trim21_Figure1.png
 <!-- Add more about the biology of this part here