This operator, when paired with a guide RNA, expresses GFP.
The 2016 BostonU iGEM team designed a set of mutually orthogonal guide RNA (gRNA) expression vectors and gRNA operator reporters that could activate the expression of a gene of interest. This part contains the gRNA operator target sequence for gRNA 8 from BostonU’s Gemini Library (5’-GTTGCGCGTCCGTATCAAGG-3’) (Part BBa_K1875006). The introduction of the gRNA leads to a significant fold increase in expression of the gene of interest in comparison to the basal level tested without the gRNA (figure 1). Figure 2 illustrates the results from a mutual orthogonality screen showing on target and off target expression levels among the guide expression vectors and the gRNA operator reporters in the Gemini Library. These results show that there is no significant expression among off target gRNA and operator pairs, demonstrating that there is no cross talk in Gemini. The Gemini system was compared to a CMV from the registry (Part BBa_I712004) to test its strength against a different promoter. This operator reporter was found to be around the same strength as the CMV; however, also from the Gemini Library, the gRNA 8 triple multimerized operator reporter was found to have much stronger expression than the CMV.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21Illegal BglII site found at 927
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000COMPATIBLE WITH RFC