Protein_Domain

Part:BBa_K2356003:Design

Designed by: Ralf Philipsen   Group: iGEM17_TU-Eindhoven   (2017-10-27)


CT33 with Strep-tag II and mCherry


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 49
    Illegal BamHI site found at 769
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 793
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 20


Design Notes

We added multiple restriction sites, so that the 3 parts may be exchanged. CT33 could for example be exchanged for a CT52 construct using SalI and SacI enzymes. We also altered the DNA sequences in order to optimize expression in E. coli. The part can easily be cloned into a pET28a(+) vector using the NcoI and SacI restriction sites.


Source

mCherry is a genomic sequence first discovered in mushroom anemones. Strep-tag II is only 8 amino acids long and experimentally proven by others to bind to Streptavidin. The CT33 part is derived from the genomic DNA coding for H+-ATPase, a protein which is present in many organisms.

References