Part:BBa_K4877022:Design
Custom DART VADAR entry sequence with HBB 5' and 3' UTRs
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 890
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 839
Illegal BamHI site found at 1143
Illegal BamHI site found at 3320
Illegal XhoI site found at 882 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 2154
Design Notes
There were several illegal restriction sites in the original sequence, which had to be edited. These were silent mutations in coding regions, but this also led to a minor change in the original Gibson assembly homology region. The additional SphI and SalI restriction sites flanking the payload sequence inevitably insert additional amino acids to the N- and C-termini of the payload protein.
Source
The part was constructed from the original published DART VADAR sequence by Gayet er al. as template [1] by introducing several changes.
[1] Gayet, R.V., Ilia, K., Razavi, S. et al. Autocatalytic base editing for RNA-responsive translational control. Nat Commun 14, 1339 (2023). https://doi.org/10.1038/s41467-023-36851-z