Part:BBa_K4697000:Design
Template for Guide RNA for Cas13 detection
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Enhancer: needed for a better transcription process, we used GAAAT, but, according to this reference, GAATT would have been a better choice Phage t7 promoters for boosting in vitro transcription. Thomas Conrad, Sascha Sauer, 2021 April 01, Google Patent, https://patents.google.com/patent/WO2021058145A1/en
T7 promoter: or any promoter for a transcription process, the sequence used is TAATACGACTCACTATAG
Scaffold for the enzyme used, as we used LwCas13a, the sequence would be GATTTAGACTACCCCAAAAACGAAGGGGACTAAAAC another possibility for example would be to change the sequence with the scaffold for CasRx, using the following sequence CCCCTACCAACTGGTCGGGGTTT
Finally, the reverse complementary sequence of the target
Source
scaffold sequence of the Cas13 enzyme, Target obtained through BLAST, using BLAST to check if the sequence is present in other organisms