Composite
Part:BBa_K2332054:Design
Designed by: Paola Handal Group: iGEM17_UCL (2017-10-21)
GFP-SpyCatcher (constitutive)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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
SpyCatcher sequence was obtained from BBa_K1159200 and GFP from: BBa_E0040. The DNA sequence was synthesised by Integrated DNA Technologies (IDT)
References
1. Zakeri B, Fierer J, Celik E, Chittock E, Schwarz-Linek U, Moy V et al. Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin. Proceedings of the National Academy of Sciences. 2012;109(12):E690-E697.