Composite

Part:BBa_K4031009

Designed by: Yuichiro Ikagawa   Group: iGEM21_Gifu   (2021-10-21)


crRNA 2 for LbCas12a with T7 promoter

We searched (TTTN), the PAM sequence of Cas12a, against T7p47 of the T7 phage and obtained the downstream sequences. we also added T7 promoter to in vitro transcription. We selected sequences with high GC rates and then performed homology searches on these sequences using nBLAST because we thought that sequences with high GC rates would have higher specificity for binding. This series of design steps led to the selection of sequences with low concordance to the human genome and to bacteria that are thought to be present in the human oral cavity and that are unlikely to act as off-targets for Cas12a.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 37
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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