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Part:BBa_J33205:Design

Designed by: Chris French   Group: iGEM06_Edinburgh   (2006-10-26)


hybrid lambda PR-lac promoter, repressed by lambda cI and lacI, plus lacZ


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 79
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part includes the entire lambda PRM-PR region, with cI binding sites OR1, OR2 and OR3 and the -35 and -10 sequences of both promoters (PRM and PR). An XhoI site was introduced by PCR, and the DNA was joined to the region of the lac promoter beginning with the LacI binding site. The distance between the LacI binding site and the PR -10 site in the fusion is approximately the same as that between the LacI binding site and its native -10 site; thus we hope that LacI will be able to repress PR in our construct. However, this has yet to be properly tested.


Source

The lambda-derived portion (bases 4 to 78) is derived from lambda DNA by PCR using primers based on published sequence (Genbank accession J02459, gi:215104). The lac portion (bases 84 to 353) is derived from E. coli BL21 genomic DNA using primers based on published sequence (Genbank accession J01636, gi:146575).

References