Difference between revisions of "Part:BBa K2361001:Design"
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===Source=== | ===Source=== | ||
| − | This part originates from the plasmid pJWV102-PL-dCas9, which is available via Addgene. this part was made biobrick compatible by removing the EcoRI-site and adding the prefix. Finally the suffix and the VRER mutations were added by replacing the last part of dCas9 with a piece of synthetic DNA. The VRER mutations were described for hCas9 by Heler et al. | + | This part originates from the plasmid pJWV102-PL-dCas9, which is available via Addgene (https://www.addgene.org/85588/). this part was made biobrick compatible by removing the EcoRI-site and adding the prefix. Finally the suffix and the VRER mutations were added by replacing the last part of dCas9 with a piece of synthetic DNA. The VRER mutations were described for hCas9 by Heler et al. |
Heler, R. et al. Mutations in Cas9 Enhance the Rate of Acquisition of Viral Spacer Sequences during the CRISPR-Cas Immune Response. Mol. Cell 65, 168–175 (2017) | Heler, R. et al. Mutations in Cas9 Enhance the Rate of Acquisition of Viral Spacer Sequences during the CRISPR-Cas Immune Response. Mol. Cell 65, 168–175 (2017) | ||
===References=== | ===References=== | ||
Revision as of 16:16, 26 October 2017
dCas9 VRER
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1100
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 3379
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
After obtaining the dCas9 in a biobrick compatible form, this was used as a backbone to create dCas9 VRER. Multiple mutations were made in the C-terminal end of the proteins sequence. These mutations were implemented by exchanging the C-terminal portion (1590bps)
Source
This part originates from the plasmid pJWV102-PL-dCas9, which is available via Addgene (https://www.addgene.org/85588/). this part was made biobrick compatible by removing the EcoRI-site and adding the prefix. Finally the suffix and the VRER mutations were added by replacing the last part of dCas9 with a piece of synthetic DNA. The VRER mutations were described for hCas9 by Heler et al.
Heler, R. et al. Mutations in Cas9 Enhance the Rate of Acquisition of Viral Spacer Sequences during the CRISPR-Cas Immune Response. Mol. Cell 65, 168–175 (2017)