Types | DnaRegion
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Roles | CDS
Coding
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Sequences | BBa_K562006_sequence (Version 1)
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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 PduT and PduU proteins from Salmonella enterica serovar Typhimurium LT2. These are components of the Salmonella bacterial microcompartment (BMC). The PduU gene product carries a C-terminal 6-His affinity/epitope tag. Production of the PduU protein has been verified by Western immunoblotting (anti-penta-His) and production of both PduT and PduU 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-TU in the Sargent Laboratory, Dundee, UK.
Notes
Therewas one native PstI sites within the pduU gene. This was 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.