Difference between revisions of "Part:BBa K2381012"
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | <h3> | + | <h3>Reference</h3> |
<p>Mitchell R. O’Connell, Benjamin L. Oakes, Samuel H. Sternberg, Alexandra East-Seletsky, Matias Kaplan & Jennifer A. Doudna (2014). Programmable RNA recognition and cleavage by CRISPR/Cas9. Nat Commun, 6, 6256. doi:10.1038/nature13769</p> | <p>Mitchell R. O’Connell, Benjamin L. Oakes, Samuel H. Sternberg, Alexandra East-Seletsky, Matias Kaplan & Jennifer A. Doudna (2014). Programmable RNA recognition and cleavage by CRISPR/Cas9. Nat Commun, 6, 6256. doi:10.1038/nature13769</p> | ||
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Revision as of 13:30, 31 October 2017
3'-OriC gRNA
It’s a gRNA plasmid which helps our CRISPR-System target to the MG1655 OriC sequence in order to achieve our expectation this year. Its backbone is taken from iGEM Kit Plate named BBa_K165005, but the BsaI restriction site which was in Amp. coding sequence has been deleted and a synonymous mutation has occurred by overlapping PCR. We used J23119 promoter, a high-intensity promoter, to obtain a higher expression of gRNA than dCas9. The 21bp-sequence comes after is a kind of sequence types taken from the article that is complementary to the Oric sequence and has a nice effect on our project. The full length gRNA scaffold is also designed after consulting literatures.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 4
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]