Difference between revisions of "Part:BBa K2332054:Design"

 
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
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To obtain the fusion protein of GFP-SpyTag, we had to remove the stop codon of GFP. Parts were already codon optimized for E. Coli.
 
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We removed the stop codon in GFP and placed the SpyCatcher sequence after a linker sequence and included a HisTag for protein purification.
  
 
===Source===
 
===Source===

Revision as of 23:46, 22 October 2017


GFP-SpyCatcher (constitutive)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 723


Design Notes

We removed the stop codon in GFP and placed the SpyCatcher sequence after a linker sequence and included a HisTag for protein purification.

Source

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 SpyCatcher: BBa_K1159200 GFPmut 3b: Part:BBa_E0040

References