Difference between revisions of "Part:BBa K3738021:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | This part is codon-optimized for <i>E. coli</i> | + | This part is codon-optimized for <i>E. coli</i>. |
The anionic tag is present to facilitate uptake into our delivery particle MS2 whereas the Histidine tag was added for the purpose of Nickel-Affinity chromatography in obtaining the purified protein. | The anionic tag is present to facilitate uptake into our delivery particle MS2 whereas the Histidine tag was added for the purpose of Nickel-Affinity chromatography in obtaining the purified protein. | ||
Latest revision as of 03:11, 20 October 2021
Lbu-Cas13a with an N-terminal Anionic Tag and C-Terminal 6XHistidine Tag
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1117
Illegal PstI site found at 1972
Illegal PstI site found at 2911 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1117
Illegal PstI site found at 1972
Illegal PstI site found at 2911 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 398
Illegal BglII site found at 1334
Illegal BglII site found at 1544
Illegal BglII site found at 1598
Illegal BglII site found at 1829
Illegal BglII site found at 2102
Illegal BglII site found at 2588 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1117
Illegal PstI site found at 1972
Illegal PstI site found at 2911 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1117
Illegal PstI site found at 1972
Illegal PstI site found at 2911
Illegal NgoMIV site found at 2050
Illegal NgoMIV site found at 2686 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
This part is codon-optimized for E. coli.
The anionic tag is present to facilitate uptake into our delivery particle MS2 whereas the Histidine tag was added for the purpose of Nickel-Affinity chromatography in obtaining the purified protein.
Source
The Cas13a protein comes from the Gram-negative bacteria Leptotrichia buccalis (Lbu).
References
Glasgow, J., Capehart, S., Francis, M., and Tullman, D (2012) Osmolyte-Mediated Encapsulation of Proteins inside MS2 Viral Capsids. ACS Nano. 10, 8658-8664.
Huynh, N., Depner, N., Larson, R. et al. A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila. Genome Biol 21, 279 (2020). https://doi.org/10.1186/s13059-020-02193-y
McDade Joel. CRISPR 101: Targeting RNA with Cas13a (C2c2). Retrieved from https://blog.addgene.org/crispr-101-targeting-rna-with-cas13a-c2c2.