These vectors were used to express eukaryotic integral membrane proteins in E. coli. They can also be used for expression of toxic proteins.
- Ref: Kim, HS et al., "Translation levels control multi-spanning membrane protein expression," PLoS One. 2012;7(4):e35844.
- pBR322 derived
- Ampicillin resistant
- Contain tRNAs for 3 rare E. coli codons (argU, glyT and pro2).
- tphac promoter= λ to transcriptional terminator upstream of a phoA/lac promoter
- Provides tight control of basal expression.
- Induction requires both phosphate starvation and addition of IPTG.
- Expression was done in host strain 58F3.
- Note: Neither construct has been sequenced completely, but based on restriction digestion results, BamHI/NotI appear to be unique and can be used for sub-cloning. Alternatively, you can use SLIC cloning
- 79 amino acid TrpLE leader (thrombin cleavable)
- EG-VEGFR1 gene
- CT FLAG + octaHis tag
- dna sequencing results