Salty_PduT

BBa_K562006 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K562006
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_PduTU BMC Components



Types
DnaRegion

Roles
CDS

Coding

Sequences BBa_K562006_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 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.

Sequence Annotation Location Component / Role(s)
BBa_K562000
synthetic RBS
pduT gene
pduU gene
Hexa-Histidine Tag
1,103
107,112
119,673
673,1020
1021,1038
feature/BioBrick engineered_region
ribosome_entry_site feature/rbs
feature/cds CDS
feature/cds CDS
sequence_feature feature/misc
igem#experience
Works
 
igem#sampleStatus
In stock
igem#status
Available
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K562006/1