Part:BBa_K323135
VioA and VioB enzymes fused with zinc fingers under pBAD promoter
The part contains two of the enzymes of the violacein biosynthetic pathway, fused to zinc finger proteins. VioA enzyme is linked to the Blues zinc finger (Part: BBa_K323085) and VioB enzyme to the Zif268 zinc finger (Part: BBa_K323134). When translated to protein, two 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 on a fixed distance.
In combination with part BBa_K323132, 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.
We have designed another DNA program (Part: BBa_K323046) with no binding site for the HivC_VioD chimeric protein. By this we showed that the program determines the final product - in our case, larger amounts of deoxyviolacein were formed.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1204
Illegal NheI site found at 2568 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
Illegal BamHI site found at 6132 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 5657
Illegal AgeI site found at 979
Illegal AgeI site found at 5853 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961
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