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OGS538

Sigma-Aldrich

PSF-TPI1-URA3 - STRONG PROMOTER YEAST PLASMID

plasmid vector for molecular cloning

Synonym(e):

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-Gew.

size 7023 bp

Bakterienauswahl

kanamycin

Replikationsursprung

2Micron
pUC (500 copies)

Peptidspaltung

no cleavage

Promoter

Promoter name: TEF1
Promoter activity: constitutive
Promoter type: yeast

Reportergen

none

Versandbedingung

ambient

Lagertemp.

−20°C

Hefeauswahl

uracil

Allgemeine Beschreibung

A strong promoter (TPI1) Saccharomyces cerevisiae yeast expression plasmid for expressing proteins of interest. The vector can be selected using the URA3 metabolite cassette that allows the plasmid to be selected for in cells deficient this gene when they are grown on media that does not contain uracil.

Promoter Expression Level: This plasmid contains the strong yeast constitutive triosephosphate isomerase (TPI1) gene promoter. It demonstrates similar levels of expression to the translation elongation factor 1 promoter.

Anwendung

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. If this is not required and you wish to use a downstream site for gene cloning you can remove the NcoI site by cleaving the plasmid with KpnI.

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.

Sequenz

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

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Lagerklassenschlüssel

12 - Non Combustible Liquids

Flammpunkt (°F)

Not applicable

Flammpunkt (°C)

Not applicable


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