Composite

Part:BBa_K3396017

Designed by: Yuxin Liu   Group: iGEM20_NUDT_CHINA   (2020-10-26)
Revision as of 00:03, 27 October 2020 by Qiuxinyuan12 (Talk | contribs)


Replaceable1-Replaceable2-P2A-Truncated Trim21-Replaceable3

CMV->Replaceable1->linker->Replaceable2->P2A->Truncated Trim21->Replaceable3->BGH

This part is a composite part providing Golden Gate assembly based rapid construction of PREDATOR Pro plasmids for controlled degradation of specific target protein.


Usage and Biology

Our favorite composite part BBa_K3396012 is a plasmid platform on which different targeting domains and protein dimerization pairs can be easily installed into the Predator Pro system. The subpart BBa_K3396007 functions as the E3 ligase to degrade target protein. The protein region BBa_K3396014 and BBa_K3396015 can be replaced into different protein dimerization pairs in order to achieve signal responsiveness. The BBa_K3396013 part can be replaced by other targeting domains (ScFv. Nanobody, ligand, etc.) to mediate targeted degradation. The parts BBa_K3396013- BBa_K3396015 were designed with flanking BsaI, BsmBI and AlwI sites for the scarless replacement of these parts.

Special Design

As a member of the collection Predator system, special designs were taken for to optimize the applicability and adaptivity of such parts. Specifically, a novel designed substitution system, through which, two proteins could be fused with their corresponding fragment at the same time using Golden-Gate Assembly, was introduced to dramatically simplify the cloning process.


Sequence and Features

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1106
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1063
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 840
    Illegal BsaI site found at 1789
    Illegal BsaI.rc site found at 786


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Parameters
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