Coding

Part:BBa_K4497014

Designed by: Till Gundlach   Group: iGEM22_Munich   (2022-10-09)


mCherry Nanobody Receptor

This part encodes the sequence for a mCherry-nanobody based receptor for secretion/ surface expression. The receptor contains a Flag- and HA-tag for immunofluorescence labeling.

Design & Cloning

Design

This part is made of the following components:

  • mCherry nanobody receptor (Part:BBa_K4497014)
    • IL-11 Signaling Sequence (Part:BBa_K4497003): Signaling Sequence for Cell surface expression
    • Flag-tag (Part:BBa_K4497012): Flag epitope tag that enables surface expression detection using immunofluorescence labeling
    • HA-tag (Part:BBa_K1150016): HA epitope tag that enables surface expression detection using immunofluorescence labeling
    • Anti-mCherry nanobody (Part:BBa_K4497002): a nanobody capable of binding mCherry

Cloning

The part is encoded on the plasmid pcDNA3.1-SP-Flag HA-Tag-mCherrynb-gp130 which was kindly provided to us by Prof. Scheller [1]

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 373
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 373
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 94
    Illegal BamHI site found at 127
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 373
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 373
  • 1000
    COMPATIBLE WITH RFC[1000]

References

[1] Mossner, S., Phan, H. T., Triller, S., Moll, J. M., Conrad, U., & Scheller, J. (2020). Multimerization strategies for efficient production and purification of highly active synthetic cytokine receptor ligands. PLOS ONE, 15(4), e0230804.


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