|Products||FRT flanked, Pro and Euk. Neomycin Selection Cassette plus loxP site(FRT-PGK-gb2-neo-FRT-loxP) from Gene Bridges GmbH|
|Company||Gene Bridges GmbH|
|Item||FRT flanked, Pro and Euk. Neomycin Selection Cassette plus loxP site(FRT-PGK-gb2-neo-FRT-loxP)|
|Description|| The FRT-PGK-gb2-neo-FRT-loxP template is designed to allow kanamycin/neomycin selection in prokaryotic and eukaryotic cells, respectively. |
It 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 generally used Tn5 promoter. The promoter of the mouse Phosphoglucokinase gene (PGK) is used as the eukaryotic promoter. A synthetic polyadenylation signal terminates the kanamycin/neomycin expression. The cassette is flanked by FRT sites for later excision by Flp recombinase.
An additional single loxP site is located at the 3 end of the cassette. Unique NotI, XhoI and HindIII 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-loxP cassette flanked by any other 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 Red/ET Recombination.
The FRT-PGK-gb2-neo-FRT-loxP template is not linear but plasmid based (3485 bp in size). Due to its R6K origin the plasmid cannot 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