Difference between revisions of "Part:BBa K2615004:Design"

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===Design Notes===
 
===Design Notes===
 
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To realize different expression level of downstream gene, according to molecular dynamics and the theory of fluctuations, we design several variants of Cys4. Ensuring that varient Cys4 can bind substrate RNA, guranteeing molecular docking, we consider the cleavage and affinity activity of Cys4. We focus on the bottom of the RNA stem, where the side-chains of Gln104 involves in two sequence-specific hydrogen bonds with the major groove faces of A19.
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We designed this part by point mutation. We changed the CAG(encoding Gln) to GCG(encoding Ala)  on the 104th site based on wild type Csy4.  
 
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===Source===
 
===Source===
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<p>
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Science Museum No.105，College of Marine Life Sciences, Ocean University of China
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  The cleavage of Cys4 releases a cis-repressive RNA module (crRNA,paired with RBS) from the masked ribosome binding site (RBS), which subsequently allows the downstream translation initiation. A Ribosome Binding Site (RBS) is an RNA sequence to which ribosomes can bind and initiate translation. Combined RBS and Cys4, it can be more convenient for other iGEMers to use this composite part without putting a RBS aequence on the upstream of Cys4 coding sequence.
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===References===
 
===References===
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[http://science.sciencemag.org/content/329/5997/1355 Haurwitz R E, Jinek M, Wiedenheft B, et al. Sequence- and structure-specific RNA processing by a CRISPR endonuclease[J]. Science, 2010, 329(5997):1355-1358.]
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Latest revision as of 13:43, 11 September 2018


Csy4-Q104A, the No.2 member of Csy4 family.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 377
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 93


Design Notes

We designed this part by point mutation. We changed the CAG(encoding Gln) to GCG(encoding Ala) on the 104th site based on wild type Csy4.

Source

Science Museum No.105，College of Marine Life Sciences, Ocean University of China

References

[http://science.sciencemag.org/content/329/5997/1355 Haurwitz R E, Jinek M, Wiedenheft B, et al. Sequence- and structure-specific RNA processing by a CRISPR endonuclease[J]. Science, 2010, 329(5997):1355-1358.]