Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Zhang Zijian & Meng Liuyi
Date created: 2014-10-05 11:00:00
Date modified: 2015-05-08 01:10:12
INPNC-Linker (rTEV)-MVN & GFP
| Types | DnaRegion |
| Roles | engineered_region Project |
| Sequences | BBa_K1378009_sequence (Version 1) |
Description
This plasmid is an additional testing plasmid for our E. coli- M. aeruginosa binding assay. It differs with the part BBa_K1378007 in that we added a special linker between INPNC and MVN. We inserted two transcription units into one plasmid and they are expressed under the promoters J23105 and J23117 respectively. After this plasmid is transformed into E. coli, the mannan binding protein MVN (Microvirin) will be displayed on the cell surface by INPNC (Ice Nucleation Protein N and C terminal) and the E. coli cell will express GFP as well. There is a linker between INPNC and MVN that can be specifically digested by rTEV protease so we can add rTEV protease to control the binding activity of our E. coli cells. The cell culture with rTEV protease added can serve as the control group for the binding assay.Notes
We optimized the sequence of MVN beforehand and chose two different promoters to avoid possible homologous recombination. To ensure the normal functioning of INPNC and MVN we also added a 10aa flexible protein domain linker in between.Source
This is a composite part so we constructed it using various different smaller parts. INPNC comes from the existing part BBa_K523013. The sequence of MVN can be found in the genomic sequence of Microcystis aeruginosa PCC7806 and the gene was synthesized by a company (sequence can be found in the part BBa_K1378003). GFP is common and comes from the existing part BBa_E0040. The amino acid sequence of the linker can be easily found in papers.| Sequence Annotation | Location | Component / Role(s) |
| J23105 Linker2 B0034 GFP B0015 Linker3 B0030 J23117 B0015 MVN INPNC | 1,35 1019,1048 44,55 1586,2305 2314,2442 986,1018 1565,1579 1522,1556 1385,1513 1049,1378 62,985 | feature/promoter promoter feature/misc sequence_feature feature/rbs ribosome_entry_site feature/cds CDS stop_codon feature/stop sequence_feature feature/misc ribosome_entry_site feature/rbs feature/promoter promoter stop_codon feature/stop feature/cds CDS feature/cds CDS |