Part:BBa_K3781013:Design
SARS-CoV-2 spike protein receptor binding domain, MocloMania B4
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 520
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 520
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 458
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 520
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 520
Illegal NgoMIV site found at 606 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
This part was directly derived from SARS-CoV2-RBD-B3_B4 by introducing a different set of MoClo overhangs via PCR. The underlying genetic template was de novo synthesized and thus easily codon optimized towards Leishmania tarentolae. Because the sequence proved to be too big for the synthesis service we wanted to make use of, we had to split the gene into an upstream and downstream region. By equipping the inner separation site with additional BbsI recognition sites and modelling the resulting overhangs to re-assemble into the functional gene, we were able to generate the intact RBD gene with a simple MoClo reaction.
Source
The amino acid sequence to this part was directly derived from the online sequence source Conserved Domain Database under CDD: cd21480
The RBD coding sequence is integrated into the whole spike protein gene that can be found in the NCBI GenBank under the Gene ID: 43740568
In nature, the receptor domain is of course part of the SARS-CoV2 RNA genome.[1]
References
- ↑ Lan, J., Ge, J., Yu, J. et al. Structure of the SARS-CoV-2 spike receptor-binding domain bound to the ACE2 receptor. Nature 581, 215–220 (2020). https://doi.org/10.1038/s41586-020-2180-5