DNA

Part:BBa_K3791010:Design

Designed by: Laura Sánchez Ruiz, Auba Fuster Palŕ   Group: iGEM21_UPF_Barcelona   (2021-10-09)


gRNA Ampicillin construct


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The repeat sequence must be placed before the spacer sequence, that is, in the 5’ extreme. It is also required to have a length between 18 and 24 nucleotides, in this case adjusting it to a consensus G-C content of approximately 50%.

Source

The repeat and the T7 promoter sequences were extracted from the parts registry (BBa_K2927006) (BBa_K1614000), whereas the spacer one was designed with the constraint that it had to be complementary to the sequence of the ampicillin-resistance gene aiming to be detected (as explained in its own part page: BBa_K3791000).

References

[1] Nguyen, L. T., Smith, B. M., & Jain, P. K. (2020). Enhancement of trans-cleavage activity of Cas12a with engineered crRNA enables amplified nucleic acid detection. Nature Communications, 11(1). https://doi.org/10.1038/s41467-020-18615-1