Part:BBa_K3329003

SpyTag003 optimized V4

This is a sub part in the SpyTag003-SpyCatcher003 system. It should be used with SpyCatcher003. It's noteworthy that, in our Protein Crosslinking design, we intended to link a triplication of SpyTag003 through elastin-like polypeptide. Moreover, SpyTag003 was expressed in both P. Pastoris and E. coli, respectively. Therefore, codon optimization must be applied in order to avoid homologous recombination and fit the chassis organism. As a result, we synthesized 9 versions of genes coding the same SpyTag003 sequence based on the codon optimization of P. Pastoris and 3 versions for the codon optimization of E. coli. Names for those 9 versions of CDS is documented from SpyTag003 optimized V1 to SpyTag003 optimized V9, in which the SpyTag003 optimized V1-V6 are for P. Pastoris, and SpyTag003 optimized V7-V9 are for E. coli.

We also built a model calculating the optimized ratio of SpyCatcher003 to SpyTag003 for gel formation. See more details in our Engineering Success webpage: https://2020.igem.org/Team:BNDS_China/Engineering

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
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]