Part:BBa_K323132
VioC, VioD and VioE enzymes fused with zinc fingers under pBAD promoter
The part contains three of the enzymes of the violacein biosynthetic pathway, fused to zinc finger proteins. VioC enzyme is linked to the Gli1 zinc finger (Part:BBa_K323086), vioD to HivC zinc finger (Part:BBa_K323087) and VioE enzyme to the PBSII zinc finger (Part:BBa_K323131). When translated to protein, three functional chimeric proteins which bind to a specific DNA sequence are produced. We designed a DNA program (Part:BBa_K323066) with these sequences to bind the chimeric enzymes to a closer proximity.
In combination with Part:BBa_K323135, we have bound all of the enzymes, necessary for synthesis of violacein, on the DNA program. This enabled a faster and increased synthesis of the desired product, as the enzymes were fixed close together and the intermediates were immediately available to the next enzyme. The latter fact also resulted in a decreased formation of an unwanted side product deoxychromoviridans.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1204
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 3078
Illegal BamHI site found at 1144 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
Illegal AgeI site found at 3177 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961
Illegal SapI.rc site found at 4320
Illegal SapI.rc site found at 4395
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