Translational_Unit

Part:BBa_K1115004:Design

Designed by: Robert Oppenheimer   Group: iGEM13_SydneyUni_Australia   (2013-09-14)

dhlA Haloalkane dehalogenase


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Primers were designed to avoid EcoRI site at the 5' end. The 5' primer included a XbaI and HindIII site for cloning with (BBa_K1115007) to form (BBa_K1115008). The 3' primer included the biobrick suffix allowing it to be directionally cloned into a standard backbone alone, or with BBa_K1115007.

Plasmid source gene contains RBS with an identical sequence to the GsiB Ribosome Binding Site (BBa_K143020) which is an endogenous B.subtilis, however the comparitive strength of this RBS is not known in E.coli.

A possible improvement on this part is the removal of this RBS to increase the flexibility of this part!

Source

Xanthobacter autotophicus EL4 isolated from Botany Bay and cloned onto pUC19 plasmid by [http://sydney.edu.au/science/people/nicholas.coleman.php Coleman Lab], School of Molecular Biosciences, University of Sydney

References

[1] Janssen, D. B. et al. Cloning of 1, 2-dichloroethane degradation genes of Xanthobacter autotrophicus GJ10 and expression and sequencing of the dhlA gene. Journal of bacteriology 171, 6791-6799 (1989).

[2] Keuning, S., Janssen, D. B. & Witholt, B. Purification and characterization of hydrolytic haloalkane dehalogenase from Xanthobacter autotrophicus GJ10. Journal of bacteriology 163, 635-639 (1985).