Part:BBa_K5127013:Design
Butyrate sensor with NOT gate (pPcha-ci-pLam-GFP)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 970
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 239
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Initially, our plasmid's RBS component, synthesized by Genscript, had an unwanted point mutation. But we did characterization to see whether it could still function well (Figure 2).
Source
The leucine-responsive regulatory protein Lrp was constitutively expressed as driven by J23101. Then, Lrp activates the expression of cI repressor in the presence of butyrate, and the cI protein inhibits promoter pLam activity (Figure 1). The pCI-Lam, PpchA, protein CI, and GFP were synthesized by Genscript. The Lrp protein was supported by NMU-China.
References
Kineret Serebrinsky-Duek, Barra, M., Danino, T., & Garrido, D. (2023). Engineered Bacteria for Short-Chain-Fatty-Acid-Repressed Expression of Biotherapeutic Molecules. Microbiology Spectrum, 11(2). https://doi.org/10.1128/spectrum.00049-23