Composite

Part:BBa_K2562005

Designed by: Marina Takehana   Group: iGEM18_ASIJ_Tokyo   (2018-09-30)


CRISPR corrected E342K mutation in SERPINA1 using an OsmY tag, a GFP reporter system, and a double t


The SERPINA1 gene coding for the Alpha-1 Antitrypsin protein is derived from Homo sapiens. The coding sequence has a point mutation at E342K, a replacement of glutamic acid for lysine. We used CRISPR technology to cut out the faulty mutation and replace the E342K point mutation with its correct amino acid in the SERPINA1 gene. Secretion was optimized through the use of an OsmY tag and reported through GFP fluorescence.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1256
    Illegal NheI site found at 1279
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 209
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1934


[edit]
Categories
//awards/composite_part
//plasmid/expression/t7
Parameters
None