Part:BBa_K1140006:Design
pTet + 37 oC RNA thermometer + mCherry (LVA)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 63
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 676
Illegal AgeI site found at 788 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The prefix sites are EcoRI and XbaI and the sufix sites are SpeI and PstI.
Source
Obtained synthetically. The RNA thermosensor sequence was obtained and adapted from [4] (u6 synthetic RNA thermometer). The mCherry coding sequence [1] was optimized for E. coli codon usage.
This composite part use the TetR sequence designed by June Rhee, Connie Tao, Ty Thomson and Louis Waldman (BBa_R0040) in 2003.
References
1. Shaner NC, Steinbach PA, and Tsien RY. (2005). A guide to choosing fluorescent proteins. Nat Methods. 2(12):905-9.
2. Elowitz MB, and Leibler S. (2000). A synthetic oscillatory network of transcriptional regulators. Nature, 20;403(6767):335-8.
3. Duval-Valentin G, and Ehrlich R. (1986). Interaction between E. coli RNA polymerase and the tetR promoter from pSC101: homologies and differences with other E. coli promoter systems from close contact point studies. Nucleic Acids Res., 14(5):1967-83.
4. Neupert J, Karcher D, Bock R. (2008). Design of simple synthetic RNA thermometers for temperature- controlled gene expression in Escherichia coli. Nucleic Acids Res, 36(19):e124.