Part:BBa_K917002
lac promoter plus RFP and sacB
This is an improved version of part BBa_K322921 (French & Kowal, 2010), which consisted of the Bacillus subtlis 168 levansucrase gene, sacB, a widely used counterselectable marker which is lethal when expressed in E. coli and other Gram negative bacteria in the presence of high sucrose concentrations (Gay, Coq, Strinmetz, Ferrari, & Hoch, 1983). In this version, the lac promoter and Red Fluorescent Protein (RFP) from BBa_J04450 (but without the terminator) have been added. The provides a controllable promoter for sacB and also a marker gene. This is valuable as sacB counter-selection is notoriously prone to escape due to loss-of-function mutations in sacB. By including RFP, we can distinguish true loss of the cassette (giving white colonies) from loss of sacB function (which should still give red colonies). This will make our experiments much easier to interpret.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 781
Illegal AgeI site found at 893 - 1000COMPATIBLE WITH RFC[1000]
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