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Key Documents

Safety Information

OGS292

Sigma-Aldrich

PSF-CMV-FMDV-HYGRO - FMDV IRES HYGROMYCIN EXPRESSION 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

form

buffered aqueous solution

mol wt

size 3929 bp

bacteria selection

kanamycin

mammalian cells selection

hygromycin

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

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packaging

pkg of 1 kit

packaging

pkg of 1 kit

packaging

pkg of 1 kit

packaging

pkg of 1 kit

manufacturer/tradename

Cytiva RPN2109

manufacturer/tradename

Cytiva RPN2209

manufacturer/tradename

Cytiva RPN2134

manufacturer/tradename

Cytiva RPN2106

color

Detection reagent 1: Colorless Detection reagent 2: Colorless

color

Detection reagent 1: Colorless Detection reagent 2: Colorless

color

Detection reagent 1: Colorless Detection reagent 2: Colorless

color

Detection reagent 1: Colorless Detection reagent 2: Colorless

storage temp.

2-8°C

storage temp.

2-8°C

storage temp.

2-8°C

storage temp.

2-8°C

General description

This vector allows the expression of two genes from one vector where the second gene produced from the mRNA is the hygromycin antibiotic resistance gene (hygro) under the control of the internal ribosome entry site (IRES) from Foot and Mouth Disease Virus (FMDV). Transcription is driven by the CMV promoter. There is a multiple cloning site immediately downstream of the CMV promoter which is then followed by the IRES element driving the hygro gene.

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

Cloning in a gene: This plasmid has been designed to be compatible with a range of cloning techniques. The multiple cloning site contains a range of standard commonly used restriction sites for cloning. Using these sites genes can be inserted using standard cloning methods with DNA ligase. Other methods such as ligase independent cloning (LIC) Gibson Assembly InFusionHD or Seamless GeneArt can also be used and because all of our plasmids are based on the same backbone the same method can be used for cloning into all of our catalogue vectors.

Multiple cloning site notes: There are a few important sites within the MCS. These include the NcoI site the XbaI site and the BsgI and BseRI sites. The NcoI site contains a start codon that is immediately downstream of both a Kozak and Shine-Dalgarno ribosomal binding site. These allow for optimal positioning of genes when the start codon is placed in this location.

The XbaI site contains a stop codon. This stop codon is positioned in a specific position in relation to the BsgI and BseRI sites that are immediately downstream. When either BseRI or BsgI cleave the plasmid they produce a TA overhang from the stop codon in the XbaI site that is compatible with all of our peptide tag plasmids cut with the same sites. BseRI and BsgI sites are non-palindromic and cleave a defined number of bases away from their binding site.

Whenever we clone a gene into our multiple cloning site we always position the start and stop codon in the same positions in the MCS. If the start and ends of the genes are not compatible with NcoI and XbaI we extend the sequence to the nearest external sites but keep the start and stop codons locations consistent.

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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Storage Class Code

12 - Non Combustible Liquids

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Regulatory Information

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