Reporter

Part:BBa_J33206:Design

Designed by: Chris French   Group: iGEM06_Edinburgh   (2006-10-26)
Revision as of 17:08, 26 October 2006 by Cfrench (Talk | contribs) (→‎References)

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Bacillus subtilis ars promoter and arsR gene plus E. coli lacZ


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 454


Design Notes

The B. subtilis ars promoter and arsR gene were amplified as a biobrick-like component, but unfortunately a base of the biobrick suffix was inadvertently omitted, so that this could not be submitted alone as a proper biobrick. The lacZ' component is similar to that submitted as BBa_J33202, but differs in including a larger amount of DNA, encoding 137 amino acid residues rather than 76. This was isolated as an unexpected minor product from the PCR reactions which gave BBa_J33202. It appears to complement the lacZ-delta-M15 mutation equally well. The two components were joined as biobricks to generate the construct given here.


Source

The B. subtilis ars promoter and arsR gene were amplified from B. subtilis 168 genomic DNA using primers based on the published genome sequence. The E. coli lacZ' was amplified from E. coli BL21 genomic DNA using primers based on published sequence data (Genbank accession J01636, gi:146575).

References

Sato, T., and Kobayashi, Y. 1998. The ars operon in the skin element of Bacillus subtilis confers resistance to arsenate and arsenite. Journal of Bacteriology 180, 1655-1661.