Composite

Part:BBa_K523025

Designed by: Chris French, Eugene Fletcher, Sylvia Ispasanie   Group: iGEM11_Edinburgh   (2011-10-28)
Revision as of 18:33, 28 October 2011 by Allancrossman (Talk | contribs)

Plac + RBS + cxnA (cex + cenA fusion)

Lac promoter and RBS, followed by Cex exoglucanase from Cellulomonas fimi fused to CenA endoglucanase from the same species. The coding sequence contains, in order:

  • The GH10 exoglucanase domain
  • A (possibly inactivated) cellulose binding domain
  • The GH6 endoglucanase domain


In the process of creating this part, we tested putative correct clones on carboxymethylcellulose (CMC) plates, with Congo red dye. This test produces a zone of clearing if CMC is degraded by endoglucanase. We had three successful colonies:


K523025 on congo red.jpg


Evidently we had three correct colonies (these had already been tested for their ability to degrade MUC, in an assay similar to that for BBa_K523016.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 1148
    Illegal NotI site found at 2846
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2759
    Illegal XhoI site found at 2257
    Illegal XhoI site found at 2506
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 781
    Illegal NgoMIV site found at 1154
    Illegal NgoMIV site found at 1656
    Illegal NgoMIV site found at 2037
    Illegal NgoMIV site found at 2962
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1201
    Illegal SapI.rc site found at 1284


[edit]
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