Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Frank Sargent
Date created: 2011-09-15 11:00:00
Date modified: 2015-05-08 01:12:41
Salty_PduAB BMC Components
| Types | DnaRegion |
| Roles | Coding CDS |
| Sequences | BBa_K562003_sequence (Version 1) |
Description
This is a composite part comprising a constituive promoter (identical to that from the basic part BBa_K562000), which is the tatABCD promoter from E. coli K-12, driving production of the PduA and PduB proteins from Salmonella enterica serovar Typhimurium LT2. These are components of the Salmonella bacterial microcompartment (BMC). The PduB gene product carries a C-terminal 6-His affinity/epitope tag. Production of the PduB protein has been verified by Western immunoblotting (anti-penta-His) and production of both PduA and PduB have been confirmed by 35S-Met labelling. The proteins have also been purified from E. coli by immobilised metal affinity chromatography (IMAC) and identified by tryptic peptide mass fingerprinting. The construct is cloned as an EcoRI / PstI fragment into pSB1C3. The clone is also known as pSB-AB in the Sargent Laboratory, Dundee, UK.Notes
There were two native PstI sites within the pduB gene. These were removed using PCR-based site-directed mutagenesis. Care was taken not to alter the primary sequence of the gene product. The fragment also carries an engineered BamHI site at its extreme 3' end.Source
Derived from Salmonella enterica serovar Typhimurium LT2 genomic sequence.| Sequence Annotation | Location | Component / Role(s) |
| BBa_K562000 synthetic RBS pduA gene pduB gene Hexa-Histidine Tag | 1,103 107,112 119,403 400,1209 1210,1227 | feature/BioBrick engineered_region feature/rbs ribosome_entry_site CDS feature/cds CDS feature/cds feature/misc sequence_feature |