|Products||FRT flanked, Pro and Euk. Neomycin Selection Cassette (FRT-PGK-gb2-neo-FRT) from Gene Bridges GmbH|
|Company||Gene Bridges GmbH|
|Item||FRT flanked, Pro and Euk. Neomycin Selection Cassette (FRT-PGK-gb2-neo-FRT)|
|Description|| The FRT-PGK-gb2-neo-FRT template is designed to allow neomycin/kanamycin selection in prokaryotic and eukaryotic cells. |
The FRT-PGK-gb2-neo-FRT template encodes the neomycin/kanamycin resistance gene which combines a prokaryotic promoter (gb2) for expression of kanamycin resistance in E.coli with a eukaryotic promoter (PGK) for expression of neomycin resistance in mammalian cells.
The prokaryotic promoter gb2 is a slightly modified version of the Em7 promoter. It mediates higher transcription efficiency than the normally used Tn5 promoter. The promoter of the mouse Phosphoglucokinase gene (PGK) is used as eukaryotic promoter. A synthetic polyadenylation signal terminates the kanamycin/neomycin expression. The cassette is flanked by FRT sites for later excision by Flp recombinase. Unique NotI and XhoI sites flank the cassette for convenient cloning with restriction sites.
Using the provided PCR template one can easily create a FRT-PGK-gb2-neo-FRT cassette flanked by any restriction sites to clone the cassette into the vector of choice. The restriction sites can be introduced by adding the corresponding sequence in the PCR primer. The template can easily be used to generate targeting constructs mediated by a single Red/ET Recombination step.
The FRT-PGK-gb2-neo-FRT template is not linear but plasmid based (3446bp in size). Due to its R6K origin it can not replicate in most E. coli strains. The PCR product can therefore be used directly for downstream applications without any further purification.
|Info|| Gene Bridges GmbH|
Im Neuenheimer Feld 584
69120 Heidekberg, Germany
Customer Service: +49 (0)6221 13708 10
Fax Number: +49 (0)6221 13708 29
Web Site: http://www.Genebridges.com