Description_x000D_ General description_x000D_ This vector contains the constant antibody coding region of the IgG heavy chain from mice (Mus musculus). It is designed to allow the seamless fusion of antibody varI`Ble regions to the constant region to create full length heavy chain antidoy expression cassettes. This is made by possible by the inclusion of a BseRI restriction site upstream from the constant region coding sequence that when cleaved produces an overhang from the first two nucleotides of the first codon of the coding sequence. This allows varI`Ble regions to be inserted by placing a BseRI site after the varI`Ble region in the correct orientation and position to generate the same overhang._x000D_ Promoter Expression Level: This plasmid contains the mammalian CMV promoter to drive gene expression. We have tested all of our mammalian promoters in a range of cell types and CMV is consistently the strongest in those we have studied. However there are many reports of the CMV promoter demonstrating silencing by methylation in long-term culture. For this reason we stock a range of other promoters that are compatible with this plasmid and are available on request._x000D_ Application_x000D_ The plasmid encodes a constant region an antibody in the main multiple cloning site positioned so that it can be cleaved to produce an overhang that allows seamless fusion with a varI`Ble region from any antibody. This allows you to create full length antibody genes with no cloning scars._x000D_ To enable this immediately upstream of the constant region coding sequence there is a BseRI restriction site. This is a type-IIS restriction enzyme that binds in one position (CAGCAG) and then cleaves a specific number of nucleotides away from the binding site regardless of the sequence at the cleavage point. We use this site in all of our antibody expression cassettes in the same position. In this plasmid cutting with BseRI will result in an overhang consisting of the first two nucleotides of the first codon of the constant region. This means that any varI`Ble region with the same overhang at its 3 prime end can be ligated into this plasmid when used in conjunction with any 5 prime site (NotI-NcoI). To add this overhang the varI`Ble region must be PCR amplified to contain any of the following sIt`s at its 3 prime end: BseRI`BsgI`BtsI or BsrDI. By using this system it allows antibody varI`Ble regions to PCR amplified and fused to any of our constant region plasmids without having to re-synthesise the entire antibody expression cassette each time._x000D_ Sequence_x000D_ Quick-reference Plasmid Map_x000D_ Please select the file type you require. For reference most cloning programs will import a .gb (Genbank) file and will show all of the plasmids features automatically when downloaded and imported._x000D_ Genebank Vector Sequence File_x000D_ FASTA Vector Sequence File_x000D_ Full Plasmid Map_x000D_ Analysis Note_x000D_ To view the Certificate of Analysis for this product, please visit www.oxfordgenetics.com._x000D_ Other Notes_x000D_ Looking for more vector options to move your experiments forward faster? Consider a custom cloning vector designed and built by Oxford Genetics™. Find out more at Oxford Genetics - Sigma's partner for cloning and expression vectors for molecular biology and synthetic biology applications._x000D_ Legal Information_x000D_ Oxford Genetics is a trademark of Oxford Genetics Ltd