This website uses cookies to ensure you get the best experience. By continuing the use this site, you agree to the use of cookies.

Please note: Your browser does not support the features used on Addgene's website. You may not be able to create an account or request plasmids through this website until you upgrade your browser. Learn more

Please note: Your browser does not fully support some of the features used on Addgene's website. If you run into any problems registering, depositing, or ordering please contact us at [email protected] Learn more

plasmid kit icon

Yeast Prototrophy Kit
(Kit # 1000000085 )

Depositing Lab:   Markus Ralser

This kit consists of 23 centromeric plasmids, derived from the pHLUM mini-chromosome, designed to restore prototrophy in common S. cerevisiae laboratory strains.

The 23 single-copy plasmids complement for deficiencies in HIS3, LEU2, URA3, MET17 or LYS2 genes and in their various combinations, to match the auxotrophic background of the popular functional-genomic yeast libraries that are based on the S288c strain.

This kit consists of a 96-well plate, and will be shipped as bacterial glycerol stocks on dry ice. Samples should be frozen at -80°C immediately upon arrival.

Add to Cart
$330 USD + shipping

Available to academics and nonprofits only.

Original Publication

Saccharomyces cerevisiae single-copy plasmids for auxotrophy compensation, multiple marker selection, and for designing metabolically cooperating communities. Mülleder M, Campbell K, Matsarskaia O, Eckerstorfer F, Ralser M. F1000Research. 2016 doi: 10.12688/f1000research.9606.1. F1000.

Description

Auxotrophic markers are useful tools in yeast cloning and genome editing, enable a large spectrum of genetic techniques, as well as facilitate the study of metabolite exchange interactions in microbial communities. If unused background auxotrophies are left uncomplemented however, the cells need to be grown in nutrient supplemented or rich growth media compositions, which precludes the analysis of biosynthetic metabolism, and which leads to a profound impact on physiology and gene expression.

The Ralser Lab created a series of single copy plasmids derived from the pHLUM minichromosome. These plasmids have a native S. cerevisiae promoter for expression of the marker genes as well as a centromeric origin. Additionally, plasmids in the pHLUM series have a fragment of the E. coli lacZ for blue-white selection.

The 23 single-copy plasmids complement for deficiencies in HIS3, LEU2, URA3, MET17 or LYS2 genes and in their various combinations, to match the auxotrophic background of the popular functional-genomic yeast libraries that are based on the S288c strain. The plasmids are further suitable for designing self-establishing metabolically cooperating (SeMeCo) communities, and possess a uniform multiple cloning site to exploit multiple parallel selection markers in protein expression experiments.

Minichromosome maps

pHLUM and pHLUK minichromosome maps for yeast vector series.

The pHLUM series expresses HIS3, LEU2, URA3, and MET17.

The pHLUK series expresses HIS3, LEU2, URA3, AND LYS2.

Series of plasmid maps in kit
Maps of the plasmids generated from pHLUM and pHLUK vector parents. If unique restriction sites were gained or lost, it is indicated in the individual plasmid map.
yeast growth assay
Yeast growth assay demonstrating prototrophy restoration with each plasmid from the pHLUM and pHLUK series.

How to Cite this Kit

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.

For your Materials and Methods section:

“The Yeast Prototrophy Kit was a gift from Markus Ralser (Addgene kit # 1000000085).”

For your Reference section:

Saccharomyces cerevisiae single-copy plasmids for auxotrophy compensation, multiple marker selection, and for designing metabolically cooperating communities. Mülleder M, Campbell K, Matsarskaia O, Eckerstorfer F, Ralser M. F1000Research 2016 doi: 10.12688/f1000research.9606.1.

Yeast Prototrophy Kit - #1000000085

Resistance Color Key

Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.

Inventory

Searchable and sortable table of all plasmids in kit. The Well column links to its kit plate map on the right. The Plasmid column links to a plasmid's individual web page.

Kit Plate Map

96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.

Resistance Color Key

Ampicillin

Inventory

Well Plasmid Resistance
A / 1 pHLUM (version 2)
Ampicillin
A / 2 pHLU
Ampicillin
A / 3 pHLM
Ampicillin
A / 4 pLUM
Ampicillin
A / 5 pHUM
Ampicillin
A / 6 pHL
Ampicillin
A / 7 pHU
Ampicillin
A / 8 pHM
Ampicillin
A / 9 pLU
Ampicillin
A / 10 pLM
Ampicillin
A / 11 pUM
Ampicillin
A / 12 pL
Ampicillin
B / 1 pH
Ampicillin
B / 2 pM
Ampicillin
B / 3 pU
Ampicillin
B / 4 pHLUK
Ampicillin
B / 5 pHLK
Ampicillin
B / 6 pHUK
Ampicillin
B / 7 pLUK
Ampicillin
B / 8 pUK
Ampicillin
B / 9 pLK
Ampicillin
B / 10 pHK
Ampicillin
B / 11 pK
Ampicillin
Data calculated @ 2017-03-25

Kit Plate Map - #1000000085

Plate #1

  1 2 3 4 5 6 7 8 9 10 11 12
A
B
C
D
E
F
G
H
Content blocked, you may need to disable your ad-blocker.