Translational_Unit

Part:BBa_K1115008:Design

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

dhlB + dhlA ligated with HindIII


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

dhlA and dhlB coding sequences with ribosome binding sites were amplified from pUC19 plasmid seperately to aviod and EcoRI site between them. HindIII sites were incorporated on the 5' ends of the fowards primer of dhlB and the reverse end of the reverse primer of dhlA. The PCR products were both digested with HindIII, XbaI and PstI and cloned into a backbone via three way ligation.

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] 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).

[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).