DNA
Part:BBa_K3791002:Design
Designed by: Laura Sánchez Ruiz, Auba Fuster Palŕ Group: iGEM21_UPF_Barcelona (2021-10-09)
Spacer gRNA Erythromycin
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
For choosing which fragment of the erythromycin-resistant gene we used for our spacer design, we had to take into account the PAM sequence needed by the nature of LbCas12a. In our case, the PAM sequence was TTTV, where V is any nucleotide except T. The PAM must be placed in the 5' extreme of the complementary chain of the one being targeted and must not appear in the target sequence. [1]
Another important consideration to bear in mind was that the spacer length was required to be between 18 and 24 nucleotides. [1]
Source
This part is the complementary sequence of an erythromycin-resistant gene DNA fragment.