Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Konstantin Borisov
Date created: 2015-09-15 11:00:00
Date modified: 2015-09-18 07:46:03
lac promoter -> RpoA-N-terminus fused to Gal4
| Types | DnaRegion |
| Roles | engineered_region Generator |
| Sequences | BBa_K1833004_sequence (Version 1) |
Description
This part encodes the fusion protein consisting of the RpoA-N-terminus and Gal4. The part has a lacUV5 promoter, so the production of protein can be induced with IPTG in any E. coli strain. RpoA is the gene encoding the alpha subunit of E. coli RNA Polymerase, and the N-terminal domain is responsible for assembly of RNAP, while the C-terminal domain is responsible for recruiting the polymerase to DNA. This fusion protein is used as part of a bacterial two-hybrid system, where the Gal4 binds to Gal11P as in the yeast 2-hybrid system.Notes
The part was designed with a lacUV5 promoter for simple induction that would not overwhelm the bacteria (as opposed to using a T7 promoter, which is often so active that it inhibits the cells from growing).Source
The protein encoded by this part is used in the PNAS 2000 paper "A bacterial two-hybrid selection system for studying protein???DNA and protein???protein interactions," vol. 97, no. 13, pgs. 7382???7387. The part submitted to the registry was synthesized de novo and thus has a different nucleotide sequence, with the coding sequence codon optimized for E. coli.| Sequence Annotation | Location | Component / Role(s) |
| lac operator lac promoter rbs start 6xHIS tag RpoA amino acids 2-235 linker Gal4 amino acids 58-97 stop terminator | 37,57 1,93 112,119 125,128 129,145 146,847 848,856 857,976 977,982 983,1070 | operator feature/operator promoter feature/promoter ribosome_entry_site feature/rbs feature/start start_codon tag feature/tag feature/protein CDS feature/protein CDS feature/protein CDS feature/stop stop_codon stem_loop feature/stem_loop |