Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Matthew Jones
Date created: 2011-09-18 11:00:00
Date modified: 2015-05-08 01:12:57
Arabinose inducible TorA Export Tagged Reflectin A1 Poly His generator
| Types | DnaRegion |
| Roles | Generator engineered_region |
| Sequences | BBa_K638401_sequence (Version 1) |
Description
This construct is a Reflectin A1 protein with a TorA signal peptide on the N terminus, which has been shown to allow export of GFP via the twin arginine pathway. In addition a poly his tag has been added to the C terminus and a pBAD promoter will control expression of the fusion protein on induction with arabinose.Notes
We used a TorA sequence from the literature that had been shown to cause GFP to be exported to the periplasm of e.coli. This sequence has also been checked for illegal restriction sites - using the tool A plasmid Editor (ApE) a restriction digest prediction revealed no sites for EcoRI, SpeI, XbaI or PstI.Source
Parts from the registry, Reflectin A1 DNA from Wendy Crookes, TorA sequence from paper: Export of active green fluorescent protein to the periplasm by the twin-arginine translocase (Tat) pathway in Escherichia coliJoanna D. Thomas1, Richard A. Daniel2, Jeff Errington2, Colin Robinson1,*
The TorA sequence was incorporated into the construct using Gibson Assembly.
| Sequence Annotation | Location | Component / Role(s) |
| BBa_I0500 AraC PC Pbad BBa_B0034 conserved BBa_K638402 BBa_K638001 reflectin A1 BBa_K128005 BBa_B0010 stem_loop BBa_B0012 T7 TE polya stop stop | 1,1210 1,1079 1043,1114 1080,1210 1219,1230 1223,1226 1237,1365 1366,2427 1366,2427 2428,2446 2454,2533 2464,2508 2542,2582 2549,2568 2569,2582 2580,2580 2444,2446 | engineered_region feature/BioBrick feature/misc sequence_feature feature/promoter promoter promoter feature/promoter feature/BioBrick engineered_region conserved_region feature/conserved engineered_region feature/BioBrick engineered_region feature/BioBrick CDS feature/cds feature/BioBrick engineered_region feature/BioBrick engineered_region feature/stem_loop stem_loop feature/BioBrick engineered_region stem_loop feature/stem_loop polyA_site feature/polya stop_codon feature/stop stop_codon feature/stop |