Coding

Part:BBa_K3406002:Design

Designed by: Xiaojie Zhou   Group: iGEM20_ZJUT_China_B   (2020-08-26)


PsmCas13b coding sequence


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 779
    Illegal BglII site found at 3113
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 157
    Illegal NgoMIV site found at 1135
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 393
    Illegal SapI.rc site found at 2124


Design Notes

The sequence of PsmCas13 protein contains rare codens,so in order to be successfully expressed,host which is is enriched in tRNAs for rare codons is required(such as E.coli Rosetta2).In addition,this protein might be toxic to the host,so pLysS is also needed to reduce it's basic expression.Meanwhile,this protein is not very soluble and should be tagged with a solubility tag, such as SUMO or MBP.


Source

Prevotella sp.MA2016

References

Gootenberg JS, Abudayyeh OO, Kellner MJ, Joung J, Collins JJ, Zhang F. Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018;360(6387):439-444. doi:10.1126/science.aaq0179