Part:BBa_K3247000
RNA Thermometer NoChill-01
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 7
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 7
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 7
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The stem-loop structure of the thermometer was created by taking the complement of the ribosome binding site (RBS) and adding nucleotides to either side. The additional bases were mutated; the resulting altered sequence is known as the variable region. Mutating the variable region keeps the RBS intact while modifying the RNA thermometer secondary structure to change the melting temperature. There is no internal BsaI cut site, since it would render the thermometer incompatible with our assembly method.
Figure 1a.Predicted RNA thermometer structure at 25 °C.
Figure 1b.Predicted RNA thermometer structure at 30 °C.
Figure 1c.Predicted RNA thermometer structure at 37 °C.
Characterization
Figure 2. Fluorescence intensity at 25 °C, 30 °C, and 37 °C.
Source
The thermometers were designed de novo using NUPACK and VSAlgorithm
References
None |