Invitrogen™ pENTR™/TEV/D-TOPO™ Cloning Kit, with One Shot™ Mach1™-T1^R Chemically Competent E. coli

Use highly efficient, 5-minute cloning strategy to directionally clone blunt-end PCR product into vector

Brand:  Invitrogen™ K253520

Product Code. 10348192

Includes: pENTR/TEV/D-TOPO vector, dNTPs, salt solution, sterile water, universal M13 sequencing primers, OneShot MachI T1 Phage resistant Chemically E. coli, S.O.C. Medium, and a pUC19 control plasmid.

Description

• Blunt-end PCR products clone directionally into pENTR/TEV/D-TOPO entry vector at greater than 90% efficiency, and include TEV protease-dependent cleavage site for removal of N-terminal tags from expressed proteins
• Rapidly shuttle cloned genes between multiple vector systems
• Reliable performance
• For access to Gateway System, PCR amplify gene of interest and add product straight to provided topoisomerase charged pENTR/SD/D-TOPO vector, incubate 5 minutes, and transform provided competent E. coli cells
• Resulting attL containing Gateway Entry clones are ready for efficient recombination with choice of vectors
• pENTR/TEV/D-TOPO vector includes Tobacco Etch Virus (TEV) recognition site for protease-dependent cleavage of an N-terminal tag from recombinant protein
• Vector has M13 and T7 primer sequencing sites and attL recombination sites flanking PCR product insertion site, allowing clones to be easily sequence verified and recombined into choice of attR containing Gateway destination vectors
• Kanamycin resistance gene and pUC origin are used for selection and high-copy propagation in E. coli
• Four base overhang on vector pairs with four base sequence designed into forward primer used in PCR reaction to provide directionality to topoisomerase ligation reaction

Cloning, Gateway Cloning

Specifications

• Cloning Method: Directional TOPO™
• No. of Reactions: 20 reactions
• Product Type: TOPO Cloning Kit
• Vector: pENTR

• Content And Storage: pENTR™⁄TEV⁄D-TOPO™ Cloning Kit contains pENTR⁄TEV⁄D-TOPO™ vector, dNTPs, salt solution, sterile water, universal M13 sequencing primers, OneShot™ MachI™ T1 Phage resistant Chemically E. coli, S.O.C. Medium, and a pUC19 control plasmid. Store Competent E. coli at -80°C. Store all other components at -20°C.
• Product Line: One Shot™
• Protein Tag: N-Terminal

For Research Use Only. Not for use in diagnostic procedures.