Device

Part:BBa_K4472991:Design

Designed by: Stine Behrmann   Group: iGEM22_Uni-Hamburg   (2022-10-11)


Split ribozyme detecting hcat and expressing GFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 331
    Illegal NheI site found at 354
    Illegal NheI site found at 686
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 588
    Illegal XhoI site found at 135
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1495


Design Notes

The promoters were to weak to get a signal in our assays, so we suggest to anyone else to use much strogner promoters. The promoters of split ribozyme half 1 and split ribozyme half 2 need to be different in order to get them synthesized. If they are the same there is too much repetition for the synthesis companies. The guideRNA should have a length of 82 basepairs (Gambill L, Staubus A, Ameruoso A, Chappell J. A split ribozyme that links detection of a native RNA to orthogonal protein outputs. bioRxiv. January 2022:2022.01.12.476080. doi:10.1101/2022.01.12.476080)



Source

The original ribozmye is from Tetrahymena thermophila. The guide RNA is complementary to the stable endogenous E.coli mRNA hcat.

References