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OGS50

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PSF-OXB20 - STRONG BACTERIAL PROMOTER 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 3857 bp

Bakterienauswahl

kanamycin

Replikationsursprung

pUC (500 copies)

Peptidspaltung

no cleavage

Promoter

Promoter name: OXB20
Promoter activity: constitutive
Promoter type: bacterial

Reportergen

none

Versandbedingung

ambient

Lagertemp.

−20°C

Allgemeine Beschreibung

Expression in bacterial cells. The RecA promoter is a very strong constitutive promoter. It is approximately 650 x stronger than the constitutive AraBAD promoter that we also sell. The RecA promoter is normally regulated by the repressor LexA. In this promoter this binding site has been ablated to enable constitutive expression.

Promoter Expression Level: This plasmid contains a constitutive bacterial promoter that does not require induction. It is the strongest bacterial promoter we sell and this can cause solubility and expression problems with some proteins. We also offer a range of other bacterial promoters that are compatible with this plasmid and are available on request.

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

Hinweis zur Analyse

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

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Lagerklassenschlüssel

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Flammpunkt (°F)

Not applicable

Flammpunkt (°C)

Not applicable


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PLoS genetics, 10(10), e1004751-e1004751 (2014-10-31)
The Notch3 signaling pathway is thought to play a critical role in cancer development, as evidenced by the Notch3 amplification and rearrangement observed in human cancers. However, the molecular mechanism by which Notch3 signaling contributes to tumorigenesis is largely unknown.
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Carcinogenesis, 37(1), 18-29 (2015-10-28)
Dickkopf-3 (Dkk-3) is a secreted protein whose expression is downregulated in many types of cancer. Endogenous Dkk-3 is required for formation of acini in 3D cultures of prostate epithelial cells, where it inhibits transforming growth factor (TGF)-β/Smad signaling. Here, we
Alexander C Cerny et al.
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Artikel

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.

A range of forward and reverse sequencing primers that allow you to sequence any insert that you make into a particular position within any plasmid. Where possible, the binding sites for each of these primers is conserved.

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