DNA
Part:BBa_K3791008:Design
Designed by: Laura Sánchez Ruiz, Auba Fuster Palŕ Group: iGEM21_UPF_Barcelona (2021-10-09)
gRNA Kanamycin
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The repeat sequence must be placed before the spacer sequence, that is, in the 5â 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 sequence was extracted from the parts registry (BBa_K2927006), whereas the spacer one was designed with the constraint that it had to be complementary to the sequence of the kanamycin-resistance gene aiming to be detected (as explained in its own part page: BBa_K3791003).
References
[1] New England Biolabs. (s/f). How do I design a guide RNA for use with EnGen Lba Cas12a? Neb.com. From: https://international.neb.com/faqs/2018/05/03/how-do-i-design-a-guide-rna-for-use-with-engen-lba-cas12a