Part:BBa_K2940006
Coding Sequence of super folding GFP with an N-terminal SpyCatcher
Coding Sequence of super fold GFP with an N-terminal SpyCatcher.
Usage and Biology
This part is used as a visual aid of the enzyme-SpyCatcher and curli fiber-SpyTag. The SpyCatcher is designed to be at the N-terminus of super fold GFP (sfGFP). A 2x GGGGS flexible linker was introduced between AzoR and SpyCatcher to make sure the domains of two proteins were not interfering with each other. SpyCatcher sequence and sfGFP sequence were acquired from iGEM registry (Part:BBa_K1650037 and Part:BBa_K515005).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 222
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 592
- 1000COMPATIBLE WITH RFC[1000]
Experimental Result
The assembled construct was transformed into BL21(DE3) strain to express recombinant protein. The colonies appeared to be green under uv light (Figure 1). The characterisation of this part need to be done in the future.
Figure 1. The GFP observation in transformation. Confirmed the activity of the sfGFP protein after linked to SpyCatcher.
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