Coding

Part:BBa_K2273015:Design

Designed by: Merle Brügmann   Group: iGEM17_TU_Dresden   (2017-09-21)


Minimal SpyCatcher codon adapted for Bacillus subtilis


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]


Design Notes

This part was generated in a modified version of RFC25, where a strong Shine Dalgarno Sequence (SD) is included, and has the following prefix and suffix:

Prefix with EcoRI, NotI, XbaI, NgoMIV and SD GAATTCGCGGCCGCTTCTAGATAAGGAGGTCAAAAATGGCCGGC
Suffix with AgeI, SpeI, NotI and PstI ACCGGTTAATACTAGTAGCGGCCGCTGCAGA

Sites of restriction enzymes generating compatible overhangs are indicated by sharing one color. (EcoRI and PstI are marked in blue, NotI in green, XbaI and SpeI in red and AgeI and NgoMIV in orange. Additionally, the Shine-Dalgarno sequence is marked in silver and the start and stop codon is underlined.)

Source

The sequence was originally designed from the fibronectin-binding protein FbaB from Streptococcus pyogenes ([http://www.uniprot.org/uniprot/Q6A1F3 Uniprot]).

References

Minimization of the SpyCatcher:

Li et. All (2013) Structural Analysis and Optimization of the Covalent Association between SpyCatcher and a Peptide Tag . J. Mol. Biol.

Codon adaption for B. subtilis:

Gilbert et. all (2017) Extracellular Self-Assembly of Functional and Tunable Protein Conjugates from Bacillus subtilis. ACS Synth. Biol.