Coding

Part:BBa_K3853009

Designed by: Peng Luo   Group: iGEM21_CPU_CHINA   (2021-09-19)
Revision as of 22:38, 12 October 2021 by Lp-tiffany (Talk | contribs)


his-tag-SpyTag-AAO

Aryl alcohol oxidase (AAO) is an enzyme containing flavin-adenine-dinucleotide (FAD). This is the version with SpyTag. In order to be able to construct our multi-enzyme complex, we introduced the SpyTag/SpyCatcher system. We added SpyTag to the N-terminus of each protein through 10 × ELP, which is a oligopeptide linker that does not affect the function of the protein it attaches to, as described in the literature. So this part can be combined with dCas9-SpyCatcher through covalent isopeptide action, thereby being immobilized on dsDNA. His-tag was added to purify the protein. We use BBa_K3853053 to construct the expression system and to express and to purify the protein.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 699
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 412
    Illegal BamHI site found at 1411
    Illegal BamHI site found at 1798
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None