Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Arunavo Chakraborty
Date created: 2016-10-12 11:00:00
Date modified: 2016-10-13 03:15:53
pBAD/araC - (N-6xHis) - Ag43 - sfGFP
| Types | DnaRegion |
| Roles | CDS Coding |
| Sequences | BBa_K2168001_sequence (Version 1) |
Description
This is a fusion protein with Ag43, a protein from E coli tagged with a 6xHis tag on the N-terminal and sfGFP tagged on C-terminal of the pro-protein. Ag43 causes cell-cell adhesion in E coli, and the 6xHis and sfGFP tags are present to allow convenient study dependence of Ag43 mediated aggregation on Ag43 expression levels, which can be determined from the sfGFP fluoresence and/or quantification of 6xHis tag. The 6xHis tag is included to allow protein concentration estimation in case the sfGFP fails to fold properly.Notes
The location for the 6xHis tag is the same as that for BBa_K2168000(Ag43 tagged with 6xHis on N-terminal under pBAD/araC promoter) and uses the same design principles. For independent folding, a flexible GGGGSGGGGS linker was added between Ag43 and sfGFP, and the linker was choosen to be not very hydrophilic or hydrophobic, since Ag43 lies very close to lipid membranes.Source
This part was made from Gibson assemblies from fragments of two previous parts - BBa_K1352000 and BBa_I746908, which code for Ag43 and sfGFP respectively.| Sequence Annotation | Location | Component / Role(s) |
| araC pBAD B0034 Ag43(PstI sites removed) (GGGGS)2 linker sfGFP Double terminator (B0015) 6xHis | 1,879 906,1190 1219,1230 1237,4371 4372,4401 4402,5118 5130,5258 1396,1413 | CDS feature/cds feature/promoter promoter ribosome_entry_site feature/rbs CDS feature/cds feature/misc sequence_feature CDS feature/cds engineered_region feature/BioBrick feature/tag tag |