Part:BBa_K2273040:Design
sfGFP with C-Terminal SpyCatcher and His Tag
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.)
References
Overkamp, W. et al. Benchmarking various green fluorescent protein variants in Bacillus subtilis, Streptococcus pneumoniae, and Lactococcus lactis for live cell imaging. Appl. Environ. Microbiol. 79, 6481–6490 (2013).
Li et. All (2013) Structural Analysis and Optimization of the Covalent Association between SpyCatcher and a Peptide Tag . J. Mol. Biol.
Zakeri et. All (2012) Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin. Applied Microbiology and Biotechnology
Gilbert et. all (2017) Extracellular Self-Assembly of Functional and Tunable Protein Conjugates from Bacillus subtilis. ACS Synth. Biol.