Difference between revisions of "Part:BBa K3128017:Design"
(→‎Source)
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T18 is from <i>bordetella pertussis</i><br>
 
T18 is from <i>bordetella pertussis</i><br>
 
Ompx is from <i> Escherichia coli</i> <br>
 
Ompx is from <i> Escherichia coli</i> <br>
−
<i> pUT18 </i> plasmid from Euromedex BACTH kit was used (containing the T25 subpart and the leucine zipper gene).<br>
+
<i> pUT18 </i> plasmid from Euromedex BACTH kit was used (containing the T18 subpart and the leucine zipper gene).<br>
 
OmpX gene and GGS linker were synthesized by IDT because they were unavailable on iGEM plates.
 
OmpX gene and GGS linker were synthesized by IDT because they were unavailable on iGEM plates.
  
 
===References===
 
===References===

Revision as of 20:35, 7 October 2019


OmpX-WT protein fused with T18 subpart of Bordetella Pertussis AC under constitutive promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1339
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 880
    Illegal NgoMIV site found at 1290
    Illegal AgeI site found at 1096
  • 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

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.

References