Part:BBa_K2541408:Design
Cold-inducible RNA thermosensor measurement device
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 639
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The pseudoknot structure is important to the function of cold-inducible RNA thermosensors. We designed the thermosensor by increasing or decreasing the base pairings and GC content in the pseudoknot.
Source
The natural cspA gene sequence is from Escherichia coli. The sequence we used is the 5'UTR of cspA. And we incrase or decrease the base pairings and GC centent of this sequence. We synthesized this sequence from a synthesis company. Other sequences are derived from iGEM Registry of Standard Biological Parts.
References
[1]Giuliodori A M, Di P F, Marzi S, et al. The cspA mRNA is a thermosensor that modulates translation of the cold-shock protein CspA.[J]. Molecular Cell, 2010, 37(1):21-33.
[2]Breaker R R. RNA Switches Out in the Cold[J]. Molecular Cell, 2010, 37(1):1-2.
[3]Kortmann J, Narberhaus F. Bacterial RNA thermometers: molecular zippers and switches.[J]. Nature Reviews Microbiology, 2012, 10(4):255-65.
[4]Pédelacq J-D, Cabantous S, Tran T, Terwilliger TC, Waldo GS. 2006. Engineering and characterization of a superfolder green fluorescent protein. Nat. Biotechnol. 24:79 –88.
[5]Overkamp W, Beilharz K, Detert O W R, et al. Benchmarking various green fluorescent protein variants in Bacillus subtilis, Streptococcus pneumoniae, and Lactococcus lactis for live cell imaging.[J]. Applied & Environmental Microbiology, 2013, 79(20):6481-6490.
[6]Segall-Shapiro T H, Sontag E D, Voigt C A. Engineered promoters enable constant gene expression at any copy number in bacteria[J]. Nature Biotechnology, 2018.