Generator

Part:BBa_K414005

Designed by: Randy Pares   Group: iGEM10_Nevada   (2010-10-14)
Revision as of 21:46, 25 October 2010 by Rpares (Talk | contribs)

ccdB minimal gene

The ccdB gene is a topoisomerase II poison and will kill most cell lines. This is an improvement to the prior ccdB cell death gene (BBa_P1010) in that the size has been reduced and it is now easier to use. CcdB can be used as a selectable marker by mixing it with the desired structure and a plasmid cut with BioBrick restriction enzymes. After ligation and transformation, because ccdB is toxic to most cell lines, the only visible colonies should contain the desired structure in the desired plasmid. CcdB is toxic to most cell lines with the exception of DB3.1 cell lines (BBa_V1005). The part has two point mutations from the original given sequence of the pK7FWG2, however it has been shown that these mutations do not affect the function of ccdB.

Sequence and Features


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 BsaI site found at 545


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Categories
Parameters
None