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OGS50

Sigma-Aldrich

PSF-OXB20 - STRONG BACTERIAL PROMOTER PLASMID

plasmid vector for molecular cloning

Synonym(s):
cloning vector, expression vector, molecular cloning vector, plasmid, plasmid vector, snapfast vector, vector
NACRES:
NA.85

form

buffered aqueous solution

mol wt

size 3857 bp

bacteria selection

kanamycin

Origin of replication

pUC (500 copies)

Peptide cleavage

no cleavage

Promoter

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

reporter gene

none

shipped in

ambient

storage temp.

−20°C

General description

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.

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. 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.

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


Regulatory Listings

Regulatory Listings are mainly provided for chemical products. Only limited information can be provided here for non-chemical products. No entry means none of the components are listed. It is the user’s obligation to ensure the safe and legal use of the product.

ISHL Indicated Name

Not applicable

ISHL Notified Names

Not applicable

Certificate of Analysis

Enter Lot Number to search for Certificate of Analysis (COA).

Certificate of Origin

Enter Lot Number to search for Certificate of Origin (COO).

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