CYPV02 is a one-step directional TOPO cloning kit that places a blunt-end PCR fragment into an enhanced pET expression vector. The insert is a fragment amplified with a high-fidelity polymerase. The reaction takes 5 minutes at room temperature and expression is under T7lac promoter control.
Catalog no. CYPV02 · For research use only
Key features
- Directional topoisomerase cloning builds the expression construct in 5 minutes at room temperature
- More than 90% of inserts are in the correct orientation, reducing screening work
- T7lac promoter gives tight control of background expression
- Ampicillin resistance marker; N-terminal and C-terminal 6x His tags aid purification
- Enterokinase (EK) site allows tag removal; for research use only
Catalog numbers and pack sizes
| Catalog # | Product | Pack size |
|---|---|---|
| CYPV0201 | pTOPO-D1 Directional pET Expression Kit | 20 reactions |
Specifications
| Vector type | pET-based directional TOPO vector |
|---|---|
| Insert | Blunt-end PCR product from a high-fidelity polymerase |
| Reaction | 5 min, room temperature |
| Orientation | >90% correct direction |
| Promoter | T7lac |
| Selection | Ampicillin |
| Tags | N- and C-terminal 6x His; EK cleavage site |
| Storage | -20 °C, 12 months |
| Use | Research use only |
Applications
- Directional cloning of blunt-end PCR fragments for E. coli expression
- Expression of His-tagged recombinant proteins
- Tag removal by enterokinase cleavage
Ordering
Single-unit orders are accepted. Quotes and invoices are in USD with payment by company bank transfer, and different items can be combined into one shipment. Send the catalog number, quantity and destination via the quote form.
pTOPO-D1 Directional pET Expression Kit FAQ
What insert does this kit need?
A blunt-end PCR fragment amplified with a high-fidelity polymerase.
How long does the cloning reaction take?
Adding the insert at room temperature for 5 minutes completes the construct.
How is the protein purified and the tag removed?
The vector adds N- and C-terminal 6x His tags for purification, and an enterokinase (EK) site allows removal of the tag.
How do I order CYPV02?
Request a quote with the catalog number; single-unit orders accepted, payment in USD by company transfer, and items can be combined into one shipment.
Browse related products: Ptopo cloning kits.