Coding

Part:BBa_K1797016

Designed by: iGEM15_Tsinghua   Group: iGEM15_Tsinghua   (2015-09-17)

dCas9-Long Linker (Gly-Gly-Ser-Gly)x3-Flp

In this part, we fuse dCas9(BBa_K1797006) and catalytic domain of Flp(BBa_K1015001) with a long linker(BBa_K243006). Flp is a site-specific recombinase. Due to its specific recognition site, the use of recombinase is limited. To solve this problem, we fuse its catalytic domain with dCas9. Then where the Flp is going to function is decided by gRNA.

Fig.1 Principles of tyrosine recombinase involved recombination(source: Mechanisms of Site-Specific Recombination. Nigel D.F. Grindley, Katrine L. Whiteson, and Phoebe A. Rice. Annual Review of Biochemistry, Vol. 75: 567 -605)

Flp is a tyrosine recombinase, which functions through a tetramer(Fig.1). That means when we use dcas9-flp fusion protein, we need to design four sgRNAs each time, which significantly increases the specificity.

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
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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