pcDNA™5/FRT/TO TOPO™ TA Expression Kit

Catalog number:  K651020

Invitrogen™  Related applications: Protein Expression

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pcDNA™5/FRT/TO, pcDNA5/FRT/TO-TOPO® and pcDNA™5/FRT/TO-E (Figure 1) are designed for targeted Flp-In™ integration followed by tetracycline-regulated expression. Each vector contains a FRT site for efficient targeted integration and a hybrid CMV/TetO2 promoter for regulated expression. In addition, pcDNA5/FRT/TO-TOPO® and pcDNA™5/FRT/TO-E are adapted for TOPO®-and Echo™ Cloning, respectively. TOPO® Cloning enables rapid ligation of PCR products into pcDNA5/FRT/TO-TOPO® in just five minutes on your bench top. Echo™ Cloning allows you to quickly shuttle your gene of interest into multiple expression vectors.
The Flp-In™ T-REx™ 293 Cell Line is available to save time in expression experiments. This cell line contains pFRT/lacZeo and pcDNA™6/TR stably integrated and is tested for its ability to regulate expression (Figure 2).
For Research Use Only. Not for use in diagnostic procedures.


Promoter: CMV⁄TO
Key Function: Regulated Expression
Protein Tag or Fusion: Untagged
Delivery Method: Transfection
Cloning Method: TOPO®-TA
Selection Agent (Eukaryotic): Hygromycin
Vector Type: TOPO-TA Vectors
Constitutive or Inducible System: Inducible
Inducing Agent: Tetracycline
Product Line: Flp-In™, T-REx™, TOPO®, pcDNA™
Product Size: 20 reactions

Contents & storage

The Flp-In™ T-REx™ Core Kit contains 20 µg each of pcDNA™5/FRT/TO, pOG44, pFRT/lacZeo, pcDNA™5/FRT/TO/CAT, and pcDNA™6/TR, 2 µg each of the CMV Forward Primer and BGH Reverse Primer, and 5 g tetracycline. Store at -20°C. The pcDNA™5/FRT/TO TOPO® TA Expression Kit contains two boxes. The TOPO® box contains 200 ng of linearized and topoisomerase I-activated vector, dNTPs, control PCR template and primers, forward and reverse primers for sequencing, and a CAT expression control. Store this box at -20°C. The One Shot® box contains transformation reagents including twenty-one 50-µl aliquots of One Shot® TOP10 Chemically Competent E. coli, S.O.C. medium, and a pUC19 supercoiled control plasmid. This box should be stored at -80°C. The pcDNA™5/FRT/TO-E Echo™ Expression Vector Kit contains 20 flg each of pcDNA™5/FRT/TO-E and pcDNA™5/FRT/TO/Uni/CAT as a positive control, 2 µg of the CMV Forward Primer, Cre recombinase, and 10X Cre recombinase buffer. Store at -20°C. All reagents are guaranteed stable for 6 months when properly stored.


Manuals & protocols