Composite

Part:BBa_K1140006:Design

Designed by: Janssel Reyes del Castillo   Group: iGEM13_UANL_Mty-Mexico   (2013-09-13)
Revision as of 22:08, 5 October 2013 by GilbertoRdz (Talk | contribs) (→‎References)

pTet + 37 oC RNA thermometer + mCherry (LVA)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Unknown
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Unknown
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 676
    Illegal AgeI site found at 788
  • 1000
    COMPATIBLE 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 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.