Part:BBa_K4472991:Design
Split ribozyme detecting hcat and expressing GFP
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE 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 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 588
Illegal XhoI site found at 135 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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.