Composite

Part:BBa_K3168004

Designed by: Eva Hanckmann, Harm van der Veer, Claire Michielsen   Group: iGEM19_TU_Eindhoven   (2019-09-12)
Revision as of 11:04, 12 September 2019 by Registry (Talk | contribs)

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dCas9-LargeBitNanoLuc

This composite part is made up of two basic parts. The first basic part (CODE) codes for a catalytically dead CRISPR associated protein 9 (dCas9). This protein binds to a specific target on double stranded DNA. This specific target is determined by the guide RNA, which makes this protein very modular. The second basic part, which is fused to dCas9, consist of a (GGS)5 linker and the large bit of NanoLuc. When the large bit of Split-NanoLuc forms a complex with the small bit in the presence of the substrate (Furimazine), blue light is emitted. dCas9-LargeBit itself is slightly bioluminescent when the substrate is added. However, the intensity is more bright when a complex is formed with the small bit. Thus this composite part is part of a Split-NanoLuc detection system, which targets a specific sequence on dsDNA and sends out a bioluminescent signal.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1099
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 3378
    Illegal BamHI site found at 4630
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//awards/composite_part/nominee
Parameters
None