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Merck

OGS528

PSF-CMV-HUKAPPA LC - HUMAN KAPPA LIGHT CHAIN ANTIBODY VECTOR

plasmid vector for molecular cloning

동의어(들):

cloning vector, expression vector, molecular cloning vector, plasmid, plasmid vector, snapfast vector, vector

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제품정보 (DICE 배송 시 비용 별도)

NACRES:
NA.85
UNSPSC Code:
12352200
Promoter:
Promoter name: CMV
Promoter activity: constitutive
Promoter type: mammalian
Origin of replication:
pUC (500 copies)
Bacteria selection:
kanamycin
Reporter gene:
none
Peptide cleavage:
no cleavage
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form

buffered aqueous solution

mol wt

size 4579 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

This is an antibody expression plasmid designed for the generation of full length human kappa light chain expression cassettes. The vector has been designed to contain a non-palindomic resitriction site called BseRI upstream from the light chain coding region. When this enzyme is used it produces an overhang from the first two two nucleotides in the first codon of the constant region. This allows seamless fusion to be made between variable regions carrying the same overhang. This can be achieved by creating a variable region PCR fragment that contains a BserRI site at the end in the opposing orientation.

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

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.

Analysis Note

To view the Certificate of Analysis for this product, please visit www.oxgene.com

Other Notes

To view sequence information for this product, please visit the product page


저장 등급

12 - Non Combustible Liquids

인화점 (F)

Not applicable

인화점 (°C)

Not applicable



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