Generator

Part:BBa_K1100009:Design

Designed by: YingQi Lou   Group: iGEM13_Fudan   (2013-09-13)
Revision as of 23:57, 27 September 2013 by Nzy (Talk | contribs) (→‎Design Notes)

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Csy4 generator with no terminator


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 448
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 164


Design Notes

There is no terminator here. Therefore, it can be add upstream of any terminator you are interested in. With a sfGFP reporter with a insulator -- the 16nt Csy4 cleavage loci, you can estimate how downstream sequence influences the terminator efficiency, like the study we did with the part BBa_K1100011. The plasmid map of K1100009.png

Source

The Csy4 part is modified from the plasmid pHMGWA-Pa14Csy4 made by Jennifer Doudna's Lab, R0040 from Kit, B0034 via PCR, Insulator: Csy4 loci via PCR.

References

Qi L, Haurwitz R E, Shao W, et al. RNA processing enables predictable programming of gene expression[J]. Nature biotechnology, 2012, 30(10): 1002-1006. 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.