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Addgene

ScEnSor Kit
(Kit # 1000000215 )

Depositing Lab:   Lisbeth Olsson

The 36 plasmid ScEnSor kit allows the user to either: (1) use just the Biosensor Module in an already-developed Saccharomyces cerevisiae strain whose intracellular environment has to be further characterized; (2) use just the Genome-Integration Module to develop a new strain introducing new expression cassettes into the genome; (3) use both modules to develop a new strain and characterize its intracellular environment.

This kit consists of one 96-well plate, and will be shipped as bacterial glycerol stocks. Individual plasmids can be ordered from each plasmid page and will be shipped as bacterial stabs.

Add to Cart
$400 USD + shipping
Available to academics and nonprofits only.

Original Publication

ScEnSor Kit for Saccharomyces cerevisiae Engineering and Biosensor-Driven Investigation of the Intracellular Environment Torello Pianale L, Olsson L. ACS Synth Biol. 2023 Aug 8. doi: 10.1021/acssynbio.3c00124. PMID: 37552581. Article

Description

The three-step (Build - Transform - Assess) ScEnSor (Saccharomyces cerevisiae Engineering + Biosensor) Kit is comprised of two modules that can be used independently or in combination.

The Genome-Integration Module comprises a set of backbone plasmids for the assembly of 1–6 transcriptional units (each consisting of a promoter, coding sequence, and terminator). The final Multi-Cassette plasmids are ready for efficient, marker-free and single-locus genome integration (in HO and/or X2 loci) using the CRISPR-Cas9 technology. Therefore, a total of 12 transcriptional units can be integrated into the genome (6 TUs per locus).

The Biosensor Module includes eight fluorescent reporters to investigate the intracellular environment of Saccharomyces cerevisiae (biosensors for ATP concentration, intracellular pH, oxidative stress, glycolytic flux, ribosome abundance, unfolded protein response, pyruvate metabolism, and ethanol consumption). The biosensor plasmids are ready to be integrated into the genome (X2 locus) using the same tools from the Genome Integration Module.

The figure consists of three columns labelled biosensor module, genome-integration module, and modules combination. In the biosensor module column there is a plasmid, plus sign, and cell, with an arrow from these to a cell with DNA inside. The genome-integration column shows four interlinked plasmids. The left, right, and top plasmids have arrows to and from a linear piece of DNA (a gene) to the left, right, and top respectively. The bottom plasmid contains a blue gene. An arrow, labelled build, points down to a single plasmid with the three genes incorporated consecutively into the bottom plasmid with the blue gene. Similar to the first column, there is a plus sign, a cell, and an arrow labelled transform to a cell containing the DNA with the three genes incorporated into one of three pieces of DNA shown in the cell. The Modules combination column is a repeat of column two with the addition of the plasmid from the first column next to the cell so that at the bottom of the column after transformation we have a cell with the blue gene on the longest strand of DNA present from the first plasmid from column one, and the three genes on the middle strand of DNA present from the built plasmid in column two, and a third shorter piece of DNA without labelled/colored genes. An arrow from all three columns labelled Assess leads down to a plot and bar graph. The plot contains four lines rising from bottom left to top right at various rates. The bar graph shows four bars in the same four colors as the plotted lines with error bars.
  • The ScEnSor Kit allows the user to either: (1) use just the Biosensor Module in an already-developed Saccharomyces cerevisiae strain whose intracellular environment has to be further characterized; (2) use just the Genome-Integration Module to develop a new strain introducing new expression cassettes into the genome; (3) use both modules to develop a new strain and characterize its intracellular environment.

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 ScEnSor Kit was a gift from Lisbeth Olsson (Addgene kit #1000000215).”

For your Reference section:

ScEnSor Kit for Saccharomyces cerevisiae Engineering and Biosensor-Driven Investigation of the Intracellular Environment Torello Pianale L, Olsson L. ACS Synth Biol. 2023 Aug 8. doi: 10.1021/acssynbio.3c00124. PMID: 37552581.

ScEnSor Kit - #1000000215

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 lists the plasmid well location in its plate. 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
Kanamycin

Inventory

Well Plasmid Resistance
A / 1 YN2_1_LT58_X2site
Ampicillin
A / 2 LT1_33_pTEFmut8-mCherry
Ampicillin
A / 3 LT1_34_sfpHluorin
Ampicillin
A / 4 LT1_36_QUEEN-2m
Ampicillin
A / 5 YN2_1_LT84_RPL13A
Ampicillin
A / 6 LT1_63_TU1of2-6_X2homology
Ampicillin
A / 7 LT1_64_TU2of2_X2homology
Ampicillin
A / 8 LT1_65_TU2of3-6_X2homology
Ampicillin
A / 9 LT1_66_TU3of3_X2homology
Ampicillin
A / 10 LT1_67_TU3of4-6_X2homology
Ampicillin
A / 11 LT1_68_TU4of4_X2homology
Ampicillin
A / 12 LT1_69_TU4of5-6_X2homology
Ampicillin
B / 1 LT1_70_TU5of5_X2homology
Ampicillin
B / 2 LT1_71_TU5of6_X2homology
Ampicillin
B / 3 LT1_72_TU6of6_X2homology
Ampicillin
B / 4 LT1_75_Donor_Tag_RPL13A-mTurq2
Ampicillin
B / 5 LT1_77_TU1of2-6_HOhomology
Ampicillin
B / 6 LT1_78_TU2of2_HOhomology
Ampicillin
B / 7 LT1_79_TU2of3-6_HOhomology
Ampicillin
B / 8 LT1_80_TU3of3_HOhomology
Ampicillin
B / 9 LT1_81_TU3of4-6_HOhomology
Ampicillin
B / 10 LT1_82_TU4of4_HOhomology
Ampicillin
B / 11 LT1_83_TU4of5-6_HOhomology
Ampicillin
B / 12 LT1_84_TU5of5_HOhomology
Ampicillin
C / 1 LT1_85_TU5of6_HOhomology
Ampicillin
C / 2 LT1_86_TU6of6_HOhomology
Ampicillin
C / 3 YN2_1_IL50_HOlocus
Ampicillin
C / 4 LT2_15_UPRpro_X2site
Kanamycin
C / 5 LT2_32_PyruEth_X2site
Kanamycin
C / 6 LT2_33_PyruPro_X2site
Kanamycin
C / 7 LT2_34_EthPro_X2site
Kanamycin
C / 8 LT1_30_backbone_X2_integration
Kanamycin
C / 9 LT1_31_backbone_HO_integration
Kanamycin
C / 10 LT2_7_OxPro
Kanamycin
C / 11 LT2_12_GlyRNA
Kanamycin
C / 12 LT2_14_GlyOx
Kanamycin
Data calculated @ 2024-11-04

Kit Plate Map - #1000000215

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