Composite
GFP-XylE

Part:BBa_K316006

Designed by: IC 2010 Team   Group: iGEM10_Imperial_College_London   (2010-10-21)
Revision as of 03:53, 26 October 2010 by Ks907 (Talk | contribs) (→‎Structure and Features)

N-terminus his tagged-GFP-XylE fusion protein


Constructed to be combined with promoter and terminator. The GFP BBa_E0040 has a 5'(5xHis) tag and is linked to XylE BBa_J33204 monomer subunit (It should be noted that the RBS of XylE (J33204) has been removed for this fusion protein). The linker is composed of a Flag tag (DYKDDDDK), TEV protease recognition sequence ENLYFQG followed by GGSGGS - flexible linker. The purpose of the linker and attached GFP is to render the XylE enzyme inactive, by preventing homo-tetramerization into its functional form.


Parts were assembled by PCR primer extension for exact methods, see our wiki [http://2010.igem.org/Team:Imperial_College_London/Strategy]


Structure and Features

GFPXylE.PNG


Sequence and Features



Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1106
    Illegal NgoMIV site found at 1278
    Illegal AgeI site found at 1629
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 662



For more information about our project please visit our wiki [http://2010.igem.org/Team:Imperial_College_London].

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