Translational_Unit

Part:BBa_K1115007:Design

Designed by: Andrew Tuckwell   Group: iGEM13_SydneyUni_Australia   (2013-09-17)

dhlB 2-haloalkanoic acid dehalogenase


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 267
    Illegal NgoMIV site found at 509
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 56


Design Notes

Primers were designed to avoid EcoRI site at the 3' end. The 3' primer included a HindIII site for cloning with (BBa_K1115004) to form (BBa_K1115008) cloned into a backbone in a three-way ligation with the standard biobrick restriciton sites.


Plasmid source gene and part contains a Shine-Dalgarno consensus sequence RBS.

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., Scheper, A., Dijkhuizen, L. & Witholt, B. Degradation of halogenated aliphatic compounds by Xanthobacter autotrophicus GJ10. Applied and environmental microbiology 49, 673-677 (1985)

[2] Van der Ploeg, J., Van Hall, G. & Janssen, D. B. Characterization of the haloacid dehalogenase from Xanthobacter autotrophicus GJ10 and sequencing of the dhlB gene. Journal of bacteriology 173, 7925-7933 (1991).