Composite

Part:BBa_K3406007:Design

Designed by: Xiaojie Zhou   Group: iGEM20_ZJUT_China_B   (2020-10-11)


6xHis-MBP-LbaCas13a


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 427
    Illegal BglII site found at 3417
    Illegal BglII site found at 4932
    Illegal BamHI site found at 1344
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2987
    Illegal AgeI site found at 4064
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1658
    Illegal BsaI.rc site found at 2741


Design Notes

The sequence of LbaCas13a protein contains rare codens,so in order to be successfully expressed,host which is enriched in tRNAs for rare codons is required(such as E.coli Rosetta2(DE3)). 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

Lachnospiraceae bacterium NK4A179

References