Part:BBa_K3128015:Design
COMP fused with T18 subpart of Bordetella Pertussis adenylate cyclase under lactose promoter
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.
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1465
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1006
Illegal NgoMIV site found at 1416
Illegal AgeI site found at 1222 - 1000COMPATIBLE WITH RFC[1000]
Source
T18 is from bordetella pertussis
Ompx is from Escherichia coli
pUT18 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.