Coding

Part:BBa_K4220008

Designed by: Xiaoya Liu   Group: iGEM22_NEU_CHINA   (2022-09-25)


P2Y2-P2A-mCherry

Introduction

The product coded by this sequence is used to show the position of P2Y2 and the function of P2A by mCherry reporter.

  • P2Y2
    • In order to increase its sensitivity and maximum response to sense eATP (extracellular ATP), we mutated K240N and F307S.[1]
  • P2A
    • After adding two proteins in the two sides of P2A, it will cleave by itself and release the two proteins at the same time.
  • mCherry
    • To report the successful cleavage of P2A peptide.

Results

1.The P2A peptide can work effectively:

caption


2.The production of P2Y2:

caption

References

1 Scott, B. M. et al. Self-tunable engineered yeast probiotics for the treatment of inflammatory bowel disease. Nature Medicine 27, 1212-1222, doi:10.1038/s41591-021-01390-x (2021).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1159
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1095
    Illegal AgeI site found at 389
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 570
    Illegal SapI site found at 615


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Categories
Parameters
None