Coding

Part:BBa_K5195012:Design

Designed by: Alice Finkelstein   Group: iGEM24_Stanford   (2024-09-29)


mNeonGreen attached to P2A, DUX4DBD, and KRAB


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 809
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 809
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 965
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 809
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 809
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 161
    Illegal SapI site found at 871
    Illegal SapI.rc site found at 133


Design Notes

The coding sequence consists of an mNeonGreen sequence, P2A linker, DUX4-DBD sequence, and KRAB repressor domain. This coding sequence was inserted into a plasmid containing an inducible Tet-On promoter, allowing for modulation of DUX4-DBD-KRAB production through the addition of varying amounts of doxycycline.


Source

mNeonGreen: bilaterian Branchiostoma lanceolatum, variant of from Aequorea victoria. P2A: porcine teschovirus 2A. DUX4-DBD: DUX4-FL, a human gene found in chromosome 4q35

References