Other

Part:BBa_K3747607:Design

Designed by: Sanne van Doorn   Group: iGEM21_Wageningen_UR   (2021-10-21)


Spacer targeting promoter Cyo oxidase


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 spacer was designed by choosing a Protospacer Adjacent Motif (PAM) sequence (5'-NGG-3'), within the non-template DNA strand sequence of the target gene. According to Batianis et al. [1], efficient gene repression could be achieved by choosing a PAM within the promoter sequence, which is why we designed the spacer to target the promoter sequence of the Cyo oxidase.

Source

Pseudomonas putida EM42

References

[1] Batianis, C., Kozaeva, E., Damalas, S. G., Martín-Pascual, M., Volke, D. C., Nikel, P. I., & Martins Dos Santos, V. (2020). An expanded CRISPRi toolbox for tunable control of gene expression in Pseudomonas putida. Microbial biotechnology, 13(2), 368–385. https://doi.org/10.1111/1751-7915.13533