Part:BBa_K4592003:Design
Synthetic RBS for Microcystis-optimized eGFP Expression
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The UTEX 2385 M. aeruginosa strain used in our project was not available as an organism for the Salis RBS Library Calculator v2.1.1 [1-3]. We instead used the PCC 7806 strain. Since the two strains have similar 16S rRNA sequences and the anti-Shine-Dalgarno sequence is found at the 3' end of the 16S rRNA sequence, we expect the estimated TIR based on the 7806 strain to be a reasonable approximation of the TIR in the UTEX 2385 strain since it is the anti-Shine-Dalgarno sequence on the ribosome that interacts with the RBS on the mRNA transcript.
Source
This synthetic RBS was generated with specified inputs into the Salis RBS Library Calculator v2.1.1. The protein CDS used was our Microcystis-optimized eGFP (BBa_K4592001). The organism used was PCC 7806 M. aeruginosa. The target minimum TIR was 5000 and target maximum TIR was 1000000.
References
[1] Reis, A. C., & Salis, H. M. (2020). An Automated Model Test System for Systematic Development and Improvement of Gene Expression Models. ACS synthetic biology, 9(11), 3145–3156. https://doi.org/10.1021/acssynbio.0c00394
[2] Farasat, I., Kushwaha, M., Collens, J., Easterbrook, M., Guido, M., & Salis, H. M. (2014). Efficient search, mapping, and optimization of multi-protein genetic systems in diverse bacteria. Molecular systems biology, 10(6), 731. https://doi.org/10.15252/msb.20134955
[3] Ng, C. Y., Farasat, I., Maranas, C. D., & Salis, H. M. (2015). Rational design of a synthetic Entner-Doudoroff pathway for improved and controllable NADPH regeneration. Metabolic engineering, 29, 86–96. https://doi.org/10.1016/j.ymben.2015.03.001