OGS568
PSF-CMV-MUKAPPA - MOUSE KAPPA LIGHT CHAIN ANTIBODY 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
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form
buffered aqueous solution
mol wt
size 4576 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-MUKAPPA – mouse kappa light chain antibody plasmid is a murine kappa light chain constant region expression plasmid. Variable antibody fragments can be fused in this plasmid to create full length antibody light chains. PSF-CMV-MUKAPPA – mouse kappa light chain antibody plasmid encodes the constant region from the mouse kappa light chain antibody gene and is designed to allow the fusion of variable antibody gene segments seamlessly to create light chain cassettes. This is possible because there is a BseRI restriction enzyme site upstream of the kappa coding sequence that when cleaved produces an overhang from the first two nucleotides of the first codon of the constant region. By designing the variable regions of antibodies to have a BseRI site at the end in an opposing direction it is possible to seamlessly fuse the variable and constant domains together.
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-MUKAPPA – mouse kappa light chain antibody 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.
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
related product
Product No.
Description
Pricing
Storage Class Code
12 - Non Combustible Liquids
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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