Regulatory
pBAD

Part:BBa_K206000:Design

Designed by: Amelia Hardjasa   Group: iGEM09_British_Columbia   (2009-10-14)
Revision as of 05:24, 18 October 2009 by Ahardjasa (Talk | contribs) (→‎Design Notes)

pBAD strong


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 65
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Niland et al. found that certain mutations in the AraI1 site increased binding of the DNA to the AraC protein. We applied all of these mutations in an attempt to create a stronger version of the pBAD promoter.

Source

Site-directed mutagenesis was performed on BBa_I13453 using the following primers:
Forward: TAATCTTATGGACTATCTTGCTATGGCATAGC
Reverse: GCGGATCCTACCTGACGCTTTTTATC

References

Niland P, Hühne R, Müller-Hill B. (1996). How AraC Interacts Specifically with its Target DNAs. J. Mol. Biol. 264:667-674. [http://www.ncbi.nlm.nih.gov/pubmed/8980677]