Part:BBa_K537001:Design
Theophylline Riboswitch 1-CheZ
- 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 effectiveness of the theophylline riboswitch is determined by the ability of the RNA 'loop' to hide the RBS preventing translation. The sequence of bases within the riboswitch will determine the formation of the loop and its leakiness. The riboswitch and its adjacent CheZ coding region are considered together and should be cloned together in order to overcome the problems previously encountered with the synthetic theophylline riboswitches submitted by teams Lethbridge 2007, Lethbridge 2009, and NYMU Taipei 2010.
Source
The cheZ sequence was PCR amplified from E.coli strain DH5α, where the forward primers in the reaction contained the sequence for the theophylline riboswitch.
The riboswitch clone 8.1 is derived from Topp and Gallivan JACS 2007, as well as previous biobricks BBa_K249026 and BBa_K411001
References
Topp S, Gallivan JP. Guiding bacteria with small molecules and RNA. J Am Chem Soc 2007;129:6807-11
Lynch S.A. Desai S.K.,Sajja,H.K., and Gallivan J.P.A high-trhoughput screen for synthetic riboswitches reveals mechanistic insight into their function. 2007, Chem Biol 14:173-184
Topp S. and Gallivan J.P. Random walks to synthetic riboswitches – a high throughput selection based on cell motility. 2008, ChemBiochem 9:210-213