Plasmid

Part:BBa_K3602020

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


Cas13a, composite part

The full Cas13a sequence (BBa_K3602001) is combined with the positively regulated lac promoter (BBa_R0011). To help translate the protein an effective ribosomal binding site (RBS) (BBa_B0034) is added. After the promoter and the RBS, the E. coli K12 optimized Cas13a protein is placed. For the terminator sequence, the part (BBa_B1006) which forms an 8 bp stem, 6 nt loop is used.

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 3059
    Illegal BglII site found at 3167
    Illegal BglII site found at 3654
    Illegal BamHI site found at 325
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1744
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 212
    Illegal BsaI.rc site found at 2896


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Categories
Parameters
None