Device

Part:BBa_K4451019:Design

Designed by: Brooks J Rady   Group: iGEM22_Sheffield   (2022-09-30)


CD + AD MutaT7 Cassette


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 74
    Illegal EcoRI site found at 3834
    Illegal PstI site found at 768
    Illegal PstI site found at 4369
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 74
    Illegal EcoRI site found at 3834
    Illegal NheI site found at 2777
    Illegal NheI site found at 2825
    Illegal NheI site found at 3737
    Illegal NheI site found at 6318
    Illegal NheI site found at 6366
    Illegal PstI site found at 768
    Illegal PstI site found at 4369
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 74
    Illegal EcoRI site found at 3834
    Illegal XhoI site found at 3755
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 74
    Illegal EcoRI site found at 3834
    Illegal PstI site found at 768
    Illegal PstI site found at 4369
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 74
    Illegal EcoRI site found at 3834
    Illegal PstI site found at 768
    Illegal PstI site found at 4369
    Illegal NgoMIV site found at 1617
    Illegal NgoMIV site found at 5158
    Illegal AgeI site found at 123
    Illegal AgeI site found at 816
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The composite part BBa_K4451019 into a low-copy pJKR-L backbone in two NEB HiFi assembly steps. We were unfortunately not able to verify the assembly of this part before the project deadline.

Source

References

Gaudelli, N. M. et al. Directed evolution of adenine base editors with increased activity and therapeutic application. Nat. Biotechnol. 38, 892–900 (2020).

Moore, C. L., Papa, L. J. & Shoulders, M. D. A Processive Protein Chimera Introduces Mutations across Defined DNA Regions In Vivo. J. Am. Chem. Soc. 140, 11560–11564 (2018).

Thuronyi, B. W. et al. Continuous evolution of base editors with expanded target compatibility and improved activity. Nat. Biotechnol. 37, 1070–1079 (2019).