Difference between revisions of "Part:BBa K4472989:Design"

 
 
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===References===
 
===References===
 +
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

Latest revision as of 03:08, 12 October 2022


Split ribozyme detecting kanamycin and expressing eforRED


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 SapI.rc site found at 41


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.



Source

The kanamycin resistance sequence for the guide RNAs was taken from pSB1K3. The original ribozmye is from Tetrahymena thermophila.

References

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