Part:BBa_K566002:Design

Biphasic switch

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 95
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

pRM and OL are spaced 2.4kb apart in Lambda where DNA naturally loops and biphasic switch works [1]. However, Electron Microscope pictures of plasmid constructions [3] show such distance may be reduced and still allow DNA looping. To our knowledge, maximum distance tested so far is 3.8kb [1].

Source

DNA synthesis.

References

1. Dodd BI, Perkins AJ, Tsemitsidis D, Egan BJ (2001) Octamerization of CI repressor is needed for effective repression of PRM and efficient switching from lysogeny Gene Dev 15:3013–3022.

2. Court DL, Oppenheim AB, Adhya LS (2007) A New Look at Bacteriophage λ Genetic Networks. J Bacteriol 189:298–304.

3. Révet B, von Wilcken-Bergmann B, Bessert H, Barker A, Müller-Hill B (1999) Four dimers of repressor bound to two suitably spaced pairs of operators form octamers and DNA loops over large distances. Curr. Biol. 9:151–154.

4. Bushman DF (1993) The bacteriophage 434 Right Operator. J. Mol. Biol. 230:28-40.

Further reading:

- Isaacs FJ, Hasty J, Cantor CR, Collins JR (2003) Prediction and measurement of an autoregulatory genetic module. Proc Natl Acad Sci USA 100:7714-7719.

- Michalowski CB, Short MD, Little JW (2004) Sequence tolerance of the phage λ PRM promoter: implications for evolution of gene regulatory circuitry. J Bacteriol 186:7899-7999.

- Voigt AC (2006) Genetic parts to program bacteria. Curr Opin Biotechnol 17:548–557.