pET28a Vectors

The pET-28a-c(+) vectors carry an N-terminal His Tag/thrombin/T7 Tag configuration plus an optional C-terminal His•Tag sequence. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circular map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-stranded sequencing should be performed using the T7 terminator primer (Cat. No. 69337-3).

The pET System is the most powerful system yet developed for the cloning and expression of recombinant proteins in E. coli. Target genes are cloned in pET plasmids under control of strong bacteriophage T7 transcription and (optionally) translation signals; expression is induced by providing a source of T7 RNA polymerase in the host cell. T7 RNA polymerase is so selective and active that, when fully induced, almost all of the cell’s resources are converted to target gene expression; the desired product can comprise more than 50% of the total cell protein a few hours after induction. All of the pET vectors and companion products are available as kits designed for convenient cloning, expression, detection, and purification of target proteins. The pET-28a-c(+) vectors carry an N-terminal His•Tag /thrombin/T7•Tag configuration plus an optional C terminal His•Tag sequence. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the vector map (TB074).

The pET Vectors are supplied as purified plasmid DNA (10 µg). Each order of pET DNA also includes an Induction Control strain (supplied as a glycerol stock). Please contact technical service if you need additional information.

This product is sold for internal research use only. Any commercial use of this product, its components, and/or any derivatives thereof (including but not limited to proteins produced using the product or its components) (together and hereinafter the ‘EMD Product’) requires the signature of a written commercial use agreement with EMD Millipore Corporation or its successor-in-interest.

Commercial use shall include but not be limited to: (1) use of the EMD Product to manufacture products for sale to third parties; (2) use of the EMD Product to provide services, information, or data to third parties in exchange for consideration; (3) use of the EMD Product for therapeutic, diagnostic or prophylactic purposes (including as part of a device, chip, assay or other product)

Or (4) resale of the EMD Product, whether or not such EMD Product is resold for research use. Nothing contained herein shall be deemed to represent or warrant that additional third-party rights are not required for use of the EMD Product. Please direct any questions on these use restrictions to: [email protected].

Hunter

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