Coding

Part:BBa_K942000

Designed by: Luis Mario Leal Garza   Group: iGEM12_Tec-Monterrey   (2012-09-26)


SS+antiHis+Etag

This part is a single chain variable fragment (scFv), this is a fusion of the variable domains of antibodies. The antigen this scFv binds to with a greater stability is a C-terminal 6xHis tag; nevertheless it binds to tags with as less as 3xHis. It is formed by two domains, the variable heavy chain domain (VH) and the variable light chain domain (VL) (Kaufmann, Lindner, & Honegger, 2002).

This protein has an estimated weight of 27.4kDa. It binds with the strongest force to C-terminal 6x His tag, but binds also to other types of His tags with different binding strength. The E-tag at the C-terminal for purification was not used in the end, instead an ultrafiltration with a 10kDa MWCO membrane was used. (Gasteiger E., 2005)

About Safety The part itself does not present any biohazard characteristic, but it is important to remember that it is designed to work in Pichia pastoris and that a biosecurity chamber is recommended to handle this organism. Its secretion to the media should not suppose any problem.

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 70
    Illegal BglII site found at 236
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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