Invitrogen™ T-REx™ Complete Kit, with pcDNA™4/TO^© Vector

Description

Includes

20μg of pcDNA4/TO or 20μg each of pcDNA4/TO/myc-His A, B, and C; a positive control vector, pcDNA;6/TR, and forward and reverse sequencing primers plus 1g Zeocin (100mg/mL), 50mg of Blasticidin, and 5g of tetracycline. All vectors are supercoiled and provided lyophilized.

Regulated mammalian expression system yields exceptionally high levels of induced expression

 

• Uses complete CMV promoter and adds control elements from bacterial tetracycline resistance operon to effectively repress and derepress transcription from one of strongest mammalian promoter sequences known
• Specific Activation: uses a repressor mechanism that blocks transcription from powerful CMV promoter in absence of tetracycline
• Because elements do not use viral transactivators, you can achieve high-level expression from complete CMV promoter without secondary, non-specific activation of host genes
• T-REx transcriptional control elements are provided through two tetracycline operator sequences (TetO₂) that have been inserted between TATA box of CMV promoter and transcriptional start site
• TetO₂ sequence itself has no effect on expression
• When the tetracycline repressor protein (TR) is present, it effectively binds TetO₂ sites and blocks transcription initiation
• Tetracycline added to culture medium binds to, and changes conformation of, TR protein; This change causes TR protein to release TetO₂ sites, derepressing transcription from CMV promoter
• Result is high-level expression of gene of interest
• Expression levels can be modulated based on tetracycline concentration and can be induced to levels that are achieved with constitutive CMV expression vectors
• T-REx is a powerful inducible mammalian expression system that allows to regulate expression from complete human cytomegalovirus (CMV) enhancer-promoter
• Complete CMV enhancer-promoter sequence containing two copies of tetracycline operator TetO₂ sequence for high-level regulated expression
• Zeocinor hygromycin resistance gene for effective selection of stable mammalian cell lines
• Large multiple cloning site to simplify cloning
• In addition, pcDNA™4/TO/myc-His offers a c-myc epitope for rapid detection of the recombinant protein with an Anti-myc Antibody and a polyhistidine (6xHis) sequence for simple purification of recombinant protein with nickel-chelating resin and detection with Anti- His(C-term) Antibody
• Regulatory vector, pcDNA™6/TR, is provided for high-level expression of the tetracycline repressor (TR) protein
• Expresses Blasticidin resistance gene for rapid selection of mammalian cell lines that stably express the TR protein

 

Mammalian Expression, Protein Expression, Proteins, Expression, Isolation and Analysis, Regulated Expression

Specifications

• Constitutive or Inducible System: Inducible
• Delivery Type: Transfection
• Inducing Agent: Tetracycline
• Promoter: CMV/TO
• Product Type: T-Rex Complete Cloning Kit with Vector
• Selection Agent (Eukaryotic): Zeocin™, Blasticidin
• Content And Storage: The T-REx™ Core Kit includes 20 μg of pcDNA™4/TO, a positive control vector, pcDNA™6/TR, and forward and reverse sequencing primers.
  
  The T-REx™ Complete Kit includes the Core Kit plus 1 g Zeocin™ (100 mg/ml), 50 mg of Blasticidin, and 5 g of tetracycline.
  
  All vectors are supercoiled and provided lyophilized.
  
  Store all components at -20°C. Zeocin™ and tetracycline should be protected from exposure to light. All components are guaranteed stable for 6 months when properly stored.
• Protein Tag: Untagged
• Cloning Method: Restriction Enzyme/MCS
• Quantity: 1 kit
• Vector: pcDNA
• Product Line: T-REx™, pcDNA™
• For Use With (Application): Regulated Expression

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