Difference between revisions of "Part:BBa K190027:Design"

(→‎Design Notes)
(→‎Design Notes)
 
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===Design Notes===
 
===Design Notes===
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Adding tobacco etch virus (TEV) cleavage site and linker site to facilitate cloning. Adding a ''Sac''I restriction site to facilitate non-expensive cloning.
+
The Maltose binding protein was fused to ArsR by the creation of a mutual linker region. The linker region contains a Tev cleavage site which helds a SacI restriction site and a string of alanine. The Tev site can be used to seperate both proteins and has been modified with a SacI restriction site for non-expensive cloning. The string of alanine residues was added to facilitate folding of the fusion protein.
  
 
===Source===
 
===Source===

Latest revision as of 12:35, 21 October 2009

MBP-ArsR fusion protein


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 360
    Illegal BglII site found at 1271
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 58
    Illegal SapI.rc site found at 1102


Design Notes

The Maltose binding protein was fused to ArsR by the creation of a mutual linker region. The linker region contains a Tev cleavage site which helds a SacI restriction site and a string of alanine. The Tev site can be used to seperate both proteins and has been modified with a SacI restriction site for non-expensive cloning. The string of alanine residues was added to facilitate folding of the fusion protein.

Source

ArsR was obtained from the E. coli K-12 DH10B genome. MBP was obtained from vector pRE-MBP (kindly provided by Guus Erkens, Membrane Enzymology group, University of Groningen).

References