Part:BBa_K3128012:Design

OmpX-WT fused with T25 subpart of Bordetella Pertussis adenylate cyclase under lactose promoter

Euromedex BACTH kit contains the pKNT25 plasmid with a MCS cassette followed by the T25 subpart of the Bordetella pertussis adenylate cyclase. It allows the cloning of the protein of interest in fusion with the T25 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.

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1636
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Source

pKNT25 plasmid from Euromedex BACTH kit was used. OmpX gene and GGS linker were synthesized by IDT because they were unavailable on iGEM plates.

References

₍₁₎ Euromedex BACTH System Kit here [http://static.bioport.cn/data/upload/product/specification/396/1342594983673_396560.pdf].