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OGS588

Sigma-Aldrich

PSF-CMV-BLAST - CMV PROMOTER BLASTICIDIN RESISTANT 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 4639 bp

bacteria selection

kanamycin

mammalian cells selection

blasticidin

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

Here the CMV promoter regulates expression of blasticidin resistance allowing simple selection of transfected mammalian cells by blasticidin selection.

Promoter Expression Level: PSF-CMV-BLAST - CMV promoter blasticidin resistant 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.

PSF-CMV-BLAST - CMV promoter blasticidin resistant plasmid is a blasticidin resistant plasmid where expression is controlled by the CMV promoter. It allows the production of mammalian cell lines carrying the plasmid.

Application

Cloning in a gene: PSF-CMV-BLAST - CMV promoter blasticidin resistant plasmid contains a gene within the main multiple cloning site (NotI-ClaI). Any plasmid that we sell where the gene is this configuration will be located in the exact same position in relation to the start and stop codon of the gene. The only exceptions to this rule are fusions proteins where the fusion gene may be positioned at the front or end of the MCS to allow gene fusion.

By positioning all of our genes in the same location it allows them to be transferred between plasmids using the same cloning method and restriction sites regardless of the plasmid being used from our product range. Inserting a new gene into this plasmid should be easily possible using a range of standard restriction enzyme sites that flank the gene currently in the molecular cloning vector.

Multiple cloning site notes: In the multiple cloning site there are two important restriction sites called BsgI and BseRI sites. These sites both cut the DNA at the same position and cleave the stop codon of the gene in the multiple cloning site in this plasmid thereby producing a TA overhang. This overhang is compatible with any of our peptide or reporter fusion tag plasmids also cut with either of these enzymes. This allows seamless C-terminal fusions to be made with the gene in this multiple cloning site using a single cloning step from our C-terminal peptide and reporter tag product range. Normally the easiest method is to clone the C-terminal tag from our other plasmid products into this plasmid using BsgI or BseRI and the downstream ClaI restriction site.

BseRI and BsgI sites are non-palindromic and cleave a defined number of bases away from their binding sites. This allows them to cut the upstream stop codon in the gene in this plasmid regardless of the gene sequence.

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

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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Articles

Learn more about relevant restriction site functions in the SnapFast™ plasmid system. All DNA sections are pre-screened, and where possible modified, to remove any of the restriction sites found within the core SnapFast plasmids to maintain their flexibility.

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