Intermediate

Part:BBa_K3128011:Design

Designed by: Melanie Dupre   Group: iGEM19_Grenoble-Alpes   (2019-09-18)
Revision as of 17:02, 18 September 2019 by Melo (Talk | contribs) (Design Notes)


OmpX-WT fused with T18 subpart of Bordetella Pertussis adenylate cyclase under lactose promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1504
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1045
    Illegal NgoMIV site found at 1455
    Illegal AgeI site found at 1261
  • 1000
    COMPATIBLE WITH RFC[1000]


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

Parts were synthesized by IDT because there were unavailable on iGEM plates.

References