Part:BBa_K3128024:Design
COMP fused with T18 subpart of Bordetella Pertussis AC under lactose promoter
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1601
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1142
Illegal NgoMIV site found at 1552
Illegal AgeI site found at 1358 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 344
Design Notes
Euromedex BACTH kit contains the pUT18 plasmid with a MCS cassette followed by the T18 subpart of the Bordetella pertussis adenylate cyclase. It allows the cloning of the protein of interest in fusion with the T18 subpart. To respect RFC rules, the plasmid has been amplificated by PCR using primers that bind from either site of the MCS cassette to eliminate it. Those primers contain also restriction sites enabling the cloning of the protein in the system. The plasmid was digested by the corresponding enzymes and the protein gene was inserted.
Source
pUT18-ZIP plasmid from Euromedex BACTH kit was used (containing the T18 subpart and the leucine zipper gene). OmpX gene and GGS linker were synthesized by IDT because they were unavailable on iGEM plates.