CV-COLRSF (Eq. pRSFDuet-1)
CV-COLRSF (Eq. pRSFDuet-1)
CV-COLRSF has a sequence identical to the pRSFDuet-1 co-expression vector. This high-copy plasmid carries a kanamycin resistance marker and an RSF1030 origin of replication, supporting stable maintenance at very high copy numbers. It includes two multiple cloning sites, each controlled by a T7 promoter and ribosome binding site, enabling robust co-expression of two target genes in E. coli.
Availability: In Production
Turn Around Time : 8 days
Format: 2 tubes with 5 µg in 5 µl of TE
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Key Properties
Applications: Bacterial Expression
Copy number: High-copy number
Expression: Constitutive
Host: E. coli
Selection markers: Kanamycin (E. coli)
Sequence: CV-COLRSF_Eq._pRSFDuet-1.gb
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Background
pRSFDuet-1 was developed as part of the Novagen Duet™ vector system to enable simultaneous expression of two genes in E. coli. CV-COLRSF preserves the exact pRSFDuet-1 sequence, ensuring compatibility with published protocols and applications. This plasmid combines a strong T7 expression system with the high-copy RSF1030 origin, allowing robust protein production. Original documentation is available in the Novagen manuals: pRSFDuet-1 Vector Manual (PDF) and related Duet Vectors Protocol TB340 (PDF).
Features
- Antibiotic selection: Kanamycin (kan)
- Origin of replication: RSF1030 (very high copy)
- Size: ~4.6 kb
- Expression system: Two T7 promoters, each with its own ribosome binding site and multiple cloning site
- Compatibility: RSF1030 origin permits co-maintenance with p15A- and ColE1-derived vectors in the same cell
Applications
- High-level co-expression of two recombinant proteins in E. coli
- Production of multi-subunit complexes where stoichiometric balance is important
- Metabolic pathway engineering requiring strong expression of key enzymes
- Parallel expression screens for protein variants
- Combination with other Duet vectors in multi-plasmid setups
Limitations
- Requires a host strain expressing T7 RNA polymerase (e.g., BL21(DE3))
- Very high copy number may place a metabolic burden on the host
- No built-in regulation beyond the T7 promoter; basal expression may occur
- Restricted to E. coli expression systems