Coding
Part:BBa_K2332032:Design
Designed by: Paola Handal Group: iGEM17_UCL (2017-10-19)
GFP-mutSpyCatcher (Lys31X, for photocaging)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 662
Design Notes
This construct was designed to not contain any other amber stop codon but at position 31 (Lys31X) (Amberless E. coli must be used to see the effects of photocageing spyCatcher). We removed the stop codon in GFP and placed the SpyTag 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)