OGS527
PSF-CMV-HUIGG1 HC - HUMAN IGG HEAVY CHAIN PLASMID
plasmid vector for molecular cloning
Synonym(s):
cloning vector, expression vector, molecular cloning vector, plasmid, plasmid vector, snapfast vector, vector
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About This Item
UNSPSC Code:
12352200
NACRES:
NA.85
form
buffered aqueous solution
mol wt
size 5248 bp
bacteria selection
kanamycin
Origin of replication
pUC (500 copies)
Peptide cleavage
no cleavage
Promoter
Promoter name: CMV
Promoter activity: constitutive
Promoter type: mammalian
reporter gene
none
shipped in
ambient
storage temp.
−20°C
General description
PSF-CMV-HUIGG1 HC - human IgG heavy chain expression plasmid contains the human IgG heavy chain constant region and has been designed to allow the seamless fusion of variable antibody fragments to create full length heavy chain expression constructs. The plasmid vector has been engineered to contain a BseRI restriction enzyme site upstream of the constant domain that when cleaved produces an overhang of the first two bases of the first codon of the constant region. This allows variable antibody fragments to be cloned upstream to create full length antibody genes. We also provide plasmids for human Kappa and Lambda light chain expression (termed pSF-CMV-HuKappa LC and pSF-CMV-HuLambda LC repectively). We also have a range of antibody expression plasmids for mouse and rat IgG heavy and light chains.
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.
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.
Application
PSF-CMV-HUIGG1 HC - human IgG heavy chain plasmid encodes a heavy chain constant region in the main multiple cloning site positioned so that it can be cleaved to produce an overhang that allows seamless fusion with a heavy chain variable region from any antibody. This allows you to create full length heavy chain 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.
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
To view sequence information for this product, please visit the product page
Analysis Note
To view the Certificate of Analysis for this product, please visit www.oxgene.com
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