DNA

Part:BBa_K3841055:Design

Designed by: Emil Ørsted Christensen   Group: iGEM21_DTU-Denmark   (2021-10-17)


Homology arm for neutral insertion site 3 in Komagataella phaffii


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 27
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 27
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 27
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 27
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The length of the homology arm is 65 base pairs as results from Frøhling, Frederik Bayer (2019) “Implementation of CRISPR-Cas9 and CASCADE Platform in Pichia Pastoris, Implementering Af CRISPR-Cas9 Og CASCADE Platformen i Pichia Pastoris.†showed that 45 base pairs was sufficient to repair DSBs in K. phaffii GS115.


Source

The homology arm is complementary to chromosome 3 (position 1457307-1457371) within the assembled genome of K. phaffii GS115 strain by De Schutter et al. 2009 (Accession number FN392320).

References