Coding

Part:BBa_K3602001

Designed by: Flora Fuglsang   Group: iGEM20_SDU-Denmark   (2020-10-23)


Cas13a w. tags, E. coli K12 optimized

This part includes the sequence for the Cas13a protein from Gootenberg et al. 2017. (https://science.sciencemag.org/content/356/6336/438). The sequence is codon-optimized for expression in E. coli K12. Upstream the gene 6x His Tag/Twin Strep/SUMO tags are placed. These can be used for his-tag purification of the protein after expression from E. coli TOP10.


Cas13a is a protein with endoribonuclease activity. In complex with a sgRNA, Cas13a cleaves nearby ssRNA upon complementary binding of a target sequence matching the sgRNA. This can be utilized for detection of different targets, as an RNA reporter can be used for the cleavage. Examples of sgRNAs fitting Cas13a are BBa_K3602002, BBa_K3602003, and BBa_K3602004.


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 2978
    Illegal BglII site found at 3086
    Illegal BglII site found at 3573
    Illegal BamHI site found at 244
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1663
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 131
    Illegal BsaI.rc site found at 2815


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Parameters
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