PSF-CMV-RAT-LAMBDA LC - RAT LAMBDA LIGHT CHAIN ANTIBODY VECTOR OGS526-5UG

NACRES:

NA.85

This cloning plasmid is designed for the production and expression of rat (Rattus rattus) lambda light chain antibody genes. The constant region of the antibody lambda gene has been positioned adjacent to a BseRI restriction site in this plasmid which when cleaved produces an overhang within the first codon of the antibody gene. This allows variable regions of antibodies to be cloned in creating seamless fusion between the the variable and constant regions. As part of this product range we can also provide expression plasmids for both human and mouse antibodies as well as cloning vectors for rat IgG heavy chain and Kappa light chain (pSF-CMV-Rat-IgG HC and pSF-CMV-Rat-Kappa respectively).
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.
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 variable region from any antibody. This allows you to create full length antibody genes with no cloning scars.
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 variable 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 variable region must be PCR amplified to contain any of the following sites at its 3 prime end: BseRI BsgI BtsI or BsrDI. By using this system it allows antibody variable 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.
Sequence
Analysis Note

form	buffered aqueous solution
mol wt	size 4570 bp
Origin of replication	pUC (500 copies)
Peptide cleavage	no cleavage
Promoter	Promoter name: CMV Promoter activity: constitutive Promoter type: mammalian
bacteria selection	kanamycin
reporter gene	none
shipped in	ambient
storage temp.	20°C

RIDADR	NONH for all modes of transport
Flash Point F	Not applicable
Flash Point C	Not applicable