Difference between revisions of "Part:BBa K2332011:Design"

 
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===Design Notes===
 
===Design Notes===
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We decided to fuse SpyCatcher to the N-terminal of a truncated version of Intimin as this has been previously proven to function effectively for cell surface display of up to 30kDa passenger proteins
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We decided to fuse SpyTag to the N-terminal of a truncated version of Intimin as this has been previously proven to function effectively for cell surface display of up to 30kDa passenger proteins (Wentzel et al., 2001). We also included a linker between Intimin and SpyTag to facilitate mobility of SpyTag on the cell surface.
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===Source===
 
===Source===

Revision as of 23:58, 22 October 2017


Blue light inducible expression of Intimin'-SpyTag (Pblind Intimin'-SpyTag)


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 880
    Illegal NgoMIV site found at 1621
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We decided to fuse SpyTag to the N-terminal of a truncated version of Intimin as this has been previously proven to function effectively for cell surface display of up to 30kDa passenger proteins (Wentzel et al., 2001). We also included a linker between Intimin and SpyTag to facilitate mobility of SpyTag on the cell surface.

Source

All parts were obtained as synthetic gBlocks Intimin protein sequence obtained from: http://www.uniprot.org/uniprot/P43261#sequences And reverse translated using: http://www.bioinformatics.org/sms2/rev_trans.html Intimin for the cell surface display of our tag was truncated according to Wentzel et al., 2001 SpyTag: BBa_K1159201 Pblind:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5001607/

References