Part:BBa_K2381002:Design
Rad1+ pMagFast
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1371
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 2214
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
In order to prevent the fusion protein has steric hindrance, We use a 12aa GS linker to link the light-induced dimerization protein with a part of dCas9, which split by a rational strategy of Cas9. According to the different function and spatial position between α-helix lobe and nuclease, we split dCas9 into two parts. Rad1 includes a-helix lobe, Rad2 includes nuclease lobe. You can use this part with https://parts.igem.org/Part:BBa_K2381004
Source
The light-induced protein nMagHigh is made from the VVD protein family derived from the fungus Neurospora crassa. The Rad1 is made from the dCas9 Protein derived from the Streptococcus pyogenes.
References
Wright A V, Sternberg S H, Taylor D W, et al. Rational design of a split-Cas9 enzyme complex.[J]. Proceedings of the National Academy of Sciences of the United States of America, 2015, 112(10):2984-2989.