Coding

Part:BBa_K4139006:Design

Designed by: Emily Hagens   Group: iGEM22_Lethbridge   (2022-10-11)
Revision as of 03:33, 12 October 2022 by Registry (Talk | contribs)

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mcyH crRNA for Lwa Cas13a (for E.coli)


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 AgeI site found at 60
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

In order to determine which region of the mcyH gene our crRNA should target, we used the RNAfold tool. This allowed us to locate a target sequence that met the ideal design criteria of being close to the 5' end of the sequence, being relatively unfolded region in the mRNA secondary structure, and having a high probability of remaining single-stranded.

This crRNA coding region was designed to be the reverse compliment of the region it will target.

Note that this crRNA will target E.coli that has been transformed with the codon-optimized mcyH gene, rather than the mcyH gene from the Microcystis aeruginosa genome.


Source

This part was designed using a segment of the mcyH gene as a reference (GenBank: AAF00956.1)

References