Difference between revisions of "Part:BBa K3406001:Design"

 
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===References===
 
===References===
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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 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15. PMID: 29449508; PMCID: PMC5961727.<br>

Latest revision as of 06:27, 27 October 2020


CcaCas13b coding sequence


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 954
    Illegal AgeI site found at 1806
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The sequence of Cas13 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,Cas13 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

Capnocytophaga canimorsus

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 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15. PMID: 29449508; PMCID: PMC5961727.