Device
AraC_14_3C

Part:BBa_K627011

Designed by: iGEM11 Potsdam_Bioware   Group: iGEM11_Potsdam_Bioware   (2011-09-21)
Revision as of 20:43, 21 September 2011 by UP Sebastian (Talk | contribs)

Fusion part of pBAD arabinose-inducible induction system and the HRV 14_3C protease
This BioBrick is a 2 part fusion of arabinose-inducible induction system and the HRV 14_3C protease. The recombinant type 14_3C protease from human rhinovirus (HRV 3C) recognizes the same cleavage site as the native enzyme: LeuGluValLeuPheGln↓GlyPro.
The small, 22 kDa size of the protease got its optimal activity at 4°C but is still very active at 37°C. It is commonly used for an easy tag removal after the purification of recombinant proteins carrying his-tag. The 14_3C works with a catalytic triade, containing the amino acid residues Ser-Asp-His at its active site.

The arabinose-inducible induction system was amplified via PCR from the pBAD_iGEMexpress vector and fused via NgoMIV with the HRV 14_3C protease. An effective induction can be accomplished with 2 mM up to 25 mM arabinose.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1523
    Illegal BglII site found at 1711
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1239
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


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Categories
Parameters
None