Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Rahul Sastry, Anjan Katta
Date created: 2013-06-09 11:00:00
Date modified: 2015-05-08 01:14:02
fnr-gal4 hybrid
| Types | DnaRegion |
| Roles | Coding CDS |
| Sequences | BBa_M36067_sequence (Version 1) |
Description
This part codes for the fnr-gal4 protein. FNR is a transcriptional activator in E. coli that is sensitive to oxygen levels. The protein is has an iron-sulfur code that, when bound to oxygen, undergoes a conformational change that reduces its ability to activate transcription. This particular coding sequence includes a nuclear localization sequence at the N-terminus (for application in eukaryotes), a gal4 activation domain (also at the N-terminus), and a stop codon.Notes
The N terminus of the sequence was chosen in order to minimize the likelihood of disrupting the protein's current function.Source
The 753-bp FNR coding sequence was isolated from K-12 DH10B E. coli, as reported on the E. coli Wiki and the Generic Genome Browser. Analysis of the 3-D structure of the protein suggested that the N-terminal would be the least interruptive location for the placement of a hybrid activation region spliced from GAL4. As such, the coding sequence for the GAL4 activating region was placed between the start and the second codons of the previously obtained FNR sequence. The coding sequence for the GAL4 activation domain was obtained from Clontech???s pGAD424 plasmid , which is used to generate hybrid GAL4 proteins for two-hybrid screens in both E. coli and S. cerevisiae. The plasmid sequence, as designed by Clontech, includes three nuclear localization signals, which were retained in the final gene design.| Sequence Annotation | Location | Component / Role(s) |
| fnr protein stop GAL4 activation domain SV40 NLS | 405,1152 1153,1155 67,405 7,63 | feature/cds CDS feature/stop stop_codon feature/cds CDS sequence_feature feature/misc |