Difference between revisions of "Part:BBa K414005"

Revision as of 20:15, 26 October 2010

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 translation 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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 <partinfo>BBa_K414005 short</partinfo>
-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.
+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 translation of ccdB.
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