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  • OGS1658 - PSF-TEF1-NH2-FLAG®-6HIS-EKT - N-TERMINAL FLAG® AND 6 HIS DUAL TAG YEAST PLASMID

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OGS1658 Sigma-Aldrich

PSF-TEF1-NH2-FLAG®-6HIS-EKT - N-TERMINAL FLAG® AND 6 HIS DUAL TAG YEAST PLASMID

plasmid vector for molecular cloning

Synonym: cloning vector, expression vector, molecular cloning vector, plasmid vector, plasmid, snapfast vector, vector

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Properties

Related Categories Cloning and Expression, Molecular Biology, SnapFast Cloning Vectors, SnapFast Vectors for Yeast Host
form   buffered aqueous solution
mol wt   size 6772 bp
conjugate   6-His tagged
  FLAG® tagged
Origin of replication   2Micron
  pUC (500 copies)
Peptide cleavage   EKT
Peptide tag location   N-terminal
Promoter   Promoter name: TEF1
Promoter activity: Constitutive
Promoter type: yeast
bacteria selection   kanamycin
reporter gene   none
shipped in   ambient
storage temp.   −20°C

Description

Analysis Note

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

General description

This plasmid is designed to express tagged proteins in yeast cells (Saccharomyces cerevisiae). The plasmid contains an auxotrophic Uracil selection expression cassette (URA3) that allows for the positive selection of yeast that are deficient in the URA3 gene (YEL021W). This is typically achieved by growing the yeast in minimal media that is reconstituted with the essential amino acids and nucleotides but excluding Uracil.

About the Peptide Tag:This plasmid contains an n-terminal Hexa-Histidine (6His) affinity tag that can be fused to a gene of interest to allow protein detection and/or purification. The sequence of the tag is: HHHHHH. This plasmid also contains a secondary Flag protein tag. The sequence of this tag is: DYKDDDDKWe provide a range of dual peptide tag plasmids. This is because some peptide tags provide specific biological properties (e. g., small molecule affinity new epitopes solubility or protein secretion) that are not provided by others.

About the Cleavage Tag: This plasmid also encodes a protease cleavage site that is designed to be positioned between your gene of interest and the tag to allow the removal of the tag following protein purification or isolation. This plasmid contains a EKT cleavage tag. The protein sequence of the cleavage tag is: DDDDK. Enterokinase (EKT) protease cleaves after the Lysine residue. It can cleave at other basic residues but this is dependent on protein confirmation. If a proline follows the site it will not cut. None of our products contain a proline after the site.

Promoter Expression Level: This plasmid contains the Yeast Elongation Factor Alpha promoter (TEF1). This is the strongest of the yeast promoters that we sell. It is a constitutive promoter and requires no induction. If you are interested in weaker promoters levels than we also stock plasmids that contain the following promoters in order of decreasing strength TPI (strong), ADHI (medium), STE5 (weak). We also stock Galactose inducible promoter plasmids if inducible expression is required. Please contact us for further information.

Legal Information

FLAG is a registered trademark of Sigma-Aldrich Co. LLC

Oxford Genetics is a trademark of Oxford Genetics Ltd

Other Notes

Looking for more vector options to move your experiments forward faster? Consider a custom cloning vector designed and built by Oxford Genetics. Find out more at Oxford Genetics - Sigma's partner for cloning and expression vectors for molecular biology and synthetic biology applications.

Sequence

Quick-reference Plasmid Map

Please select the file type you require. For reference most cloning programs will import a .gb (Genbank) file and will show all of the plasmids features automatically when downloaded and imported.

Genebank Vector Sequence File

FASTA Vector Sequence File

Full Plasmid Map

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis

Protocols & Articles

Articles

Genome Compiler

Sigma® Life Science is using our strengths and building on the strengths of others to provide the best tools to synthetic biology researchers. We are happy to participate in Genome Compiler's free, w...
Keywords: Cloning

How to Choose a SnapFast™ Expression Vector

With >1600 vectors to choose from, you have many options when planning your cloning constructs. Finding the perfect expression vector starts with understanding what you need, and what the options are.
Keywords: Cloning, Gene expression, Genetics, Immunoprecipitation, Individual protein Immunoprecipitation, Methylations, Purification, Western blot

Plasmid Product Nomenclature

The SnapFast system is a versatile plasmid cloning platform that provides a range of functional DNA sequences in an easy to clone format. We currently have hundreds of pre-designed DNA sections that ...
Keywords: Catalysis, Cloning, Digestions, PAGE

Primers for Plasmid Sequencing

We have designed a range of forward and reverse sequencing primers that allow you to sequence any insert that you make into a particular position within any of our SnapFast™ expression vectors. Other...
Keywords: Cloning, Gas chromatography, Gene expression, Melting, Sequencing

Restriction Site Positions and Functions

Click on the relevant restriction site to learn more about its function in the SnapFast plasmid system.
Keywords: Antibiotics, Cloning, Diagnostic, Gene expression, PAGE, Peptide synthesis, Phosphorylations, Polymerase chain reaction, Purification, Recombination, Sequencing

SnapFast™ Cloning & Expression Vectors Selection Charts

With over 1600 vectors to choose from, your options may seem endless. For help choosing the right vector for your research needs, consult the tables below.
Keywords: Cloning, Gene expression, Genetics, Molecular biology, Transcription

What is a DNA Component?

Our plasmids are composed of both a core vector backbone and a series of DNA components. Our plasmid platform is all built around the same core backbone, which means that the DNA components within ea...
Keywords: Adsorption, Antibiotics, Apoptosis, Cell culture, Cell disruption, Centrifugation, Chromatography, Cloning, Dialysis, Enzyme activity, Flow cytometry, Gene expression, Genetics, Immobilization, Immunoprecipitation, Nucleic acid denaturation, Precipitation, Purification, Transcription, Transfection, Western blot

Protocols

Cloning the Gene-of-Interest into a Plasmid Vector

Genetic engineering is used in thousands of laboratories around the world. Given its importance it is remarkable that cloning strategies for many of the popular DNA components are not standardised. C...
Keywords: Antibiotics, Buffers, Cloning, Degradations, Dialysis, Digestions, Gene expression, Genetic, Genetics, Individual protein Immunoprecipitation, Molecular biology, Nucleic acid annealing, Peptide synthesis, Phosphorylations, Polymerase chain reaction, Precipitation, Purification, Sequencing, Substitutions, transformation

Quick Start Plasmid Vector Guide

SnapFast™ Vectors are supplied in solution as 5µg plasmid DNA in TE buffer, at ambient temperature. If you are not ready to use your DNA right away, store this in the -20 °C for long-term storage, or...
Keywords: Cloning, Genetics, PAGE, Purification, Transfection, transformation

Restriction Enzyme Cloning Manual

Molecular Cloning is a set of techniques that are used by molecular biologists to insert a gene-of-interest into a vector capable of replication within the target cell. Once the gene is being express...
Keywords: AGE, Cloning, Digestions, Electrophoresis, Gel electrophoresis, Gene expression, Genetic, Genetics, Molecular biology, Molecular biology techniques, Nucleic acid annealing, Peptide synthesis, Phase transitions, Phosphorylations, Polymerase chain reaction, Precipitation, transformation

Selecting Correctly Expressing Recombinants

Blue / White colony screening is a strategy to quickly and easily distinguish between recombinant and non-recombinant colonies. It requires a special vector and a special strain of E. coli. It is par...
Keywords: AGE, Amplification, Antibiotics, Cell disruption, Centrifugation, Cloning, Degradations, Detergents, Diagnostic, Digestions, Electrophoresis, Gel electrophoresis, Molecular biology, Peptide synthesis, Phase transitions, Polymerase chain reaction, Precipitation, Purification, Sequencing, Transfection, transformation

SnapFast™ Cloning & Expression Vectors FAQs

Where do I find the sequences and maps of a SnapFast™ plasmid and insert? How can I confirm the orientation of my insert if I use only one insertion site? How have you removed unwanted restriction si...
Keywords: Buffers, Cloning, Diagnostic, Digestions, Gene expression, Genetics, PAGE, Peptide synthesis, Polymerase chain reaction, Sequencing, transformation

Transforming E. coli with Engineered Plasmid

Inoue and colleagues developed this method in 1990. It works well for many strains commonly used in cloning. The original method calls for growing the overnight E. coli cultures at 18°C. We find that...
Keywords: Antibiotics, Centrifugation, Cloning, Peptide synthesis, Phase transitions, transformation

Related Content

Synthetic Biology Resources

We are using our stregnths to help build the field of Synthetic Biology. Below is a collection of links to our favorite resources. In addtion to these resources, we also offer thousands of products f...
Keywords: Cell culture, Cloning, Gene expression, Genetic, Genetics, Metabolic Pathways, Metabolites, Molecular biology, Molecular biology techniques, Photosynthesis, Poisons, Polymerase chain reaction - quantitative, Transcription, Transfection

Peer-Reviewed Papers
15

References

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