Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Konstantin Borisov
Date created: 2015-09-15 11:00:00
Date modified: 2015-09-16 08:28:23
pProt-GFP
| Types | DnaRegion |
| Roles | Reporter engineered_region |
| Sequences | BBa_K1833003_sequence (Version 1) |
Description
This part produces GFP in the presence of sigma-70 E. coli RNA polymerase; however, the promoter contains two sites for a synthetic repressor to bind. Without the presence of this repressor, the part can serve as a strong constitutive promoter. For more information on the repressor, visit http://2015.igem.org/Team:Pitt/Protease/Project.This part is similar in function to BBa_K1833001, but was synthesized de novo. As such, it contains several unique features. First of all, the sequence for GFP was codon optimized. Secondly, the RBS was optimized for high translational activity using the Salis Lab Ribosome Binding Site calculator (https://salislab.net/software/).
Notes
This part was designed to be used in a cell-free system. As such, we desired the reporter to be transcribed and translated quickly. The promoter was chosen to be a strong sigma-70 E. coli promoter, with the binding sites of the repressor added between the -35 and -10 consensus sequence. Then, the RBS was optimized using the Salis Lab Ribosome Binding Site calculator (https://salislab.net/software/). The combination of the two was used to increase efficiency of in vitro transcription and translation.Source
This part was synthesized by IDT in two parts, which were then cloned together and into pSB1C3 using Gibson assembly.| Sequence Annotation | Location | Component / Role(s) |
| pProt OR2 OR1 Optimized RBS GFP start stop terminator | 1,48 7,23 29,48 49,83 84,801 81,84 798,801 805,889 | promoter feature/promoter feature/operator operator feature/operator operator feature/rbs ribosome_entry_site feature/cds CDS start_codon feature/start stop_codon feature/stop stem_loop feature/stem_loop |