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Marionette Sensor Collection
(Kit # 1000000137 )

Depositing Lab:   Christopher Voigt

The Marionette Sensor Collection enables the modular control of up to 12 genes, simply by placing each one under the control of a small (50 to 289 base pair) inducible promoter. This means that a single construct can be built and then the expression levels perturbed in many ways through the combination of different small molecules.

This kit will be sent as bacterial glycerol stocks in 96-well plate format.

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$300 USD + shipping

Available to academics and nonprofits only.

Original Publication

Escherichia coli “Marionette” strains with 12 highly optimized small-molecule sensors. Meyer AJ, Segall-Shapiro TH, Glassey E, Zhang J, and Voigt CA. Nat Chem Biol. 2019; doi: 10.1038/s41589-018-0168-3. PubMed PMID 30478458.

Description

The Marionette Sensor Collection consists of three strains and 26 plasmids:

  • The three new “Marionette” strains (sAJM.1504, sAJM.1505, and sAJM.1506) contain the Marionette cluster, which expresses 12 inducible transcription factors, from the genome. These “Marionette” strains allow for the independent control of gene expression using 2,4-diacetylphophloroglucinol (DAPG), cuminic acid (Cuma), 3-oxohexanoyl-homoserine lactone (OC6), vanillic acid (Van), isopropyl beta-D-1-thiogalactopyranoside (IPTG), anhydrotetracycline (aTc), L-arabinose (Ara), choline chloride (Cho),naringenin (Nar), 3,4-dihydroxybenzoic acid (DHBA), sodium salicylate (Sal), and 3-hydroxytetradecanoyl-homoserine lactone (OHC14).

  • Twelve plasmids (pAJM.711, pAJM.712, pAJM.713, pAJM.714, pAJM.715, pAJM.717, pAJM.716, pAJM.718, pAJM.719, pAJM.1459, pAJM.721, and pAJM.944) are reporters for those 12 transcription factors. For example: TetR is being expressed from the genome of the Marionette strains, and pAJM.717 has a promoter that is repressed by TetR (P Tet* ) driving YFP. pAJM.717 should not produce YFP in a Marionette strain, until the cell is exposed to anhydrotetracycline (aTc).

  • The other 14 plasmids (pAJM.847, pAJM.657, pAJM.474, pAJM.773,pAJM.336,pAJM.011,pAJM.677, pAJM. 683, pAJM.661, pAJM.690, pAJM.771, pAJM.1642, pAJM.884, and pAJM.969) encode both the reporter and the transcription factor, and should be used in a regular (non-Marionette) E. coli strain. For example: pAJM.011 expresses TetR and contains a P Tet*-YFP reporter module.

Marionette_Kit,_Voigt_Lab;_non_copyrighted_image.png

Figure Legend: A 12.6 kb insert expressing 12 small-molecule inducible transcription factors ("sensors") has been inserted into the glvC locus of three strains of E. coli (MG1655, DH10B, and BL21). Each sensor controls transcription from its own output promoter, which can be introduced on a medium copy plasmid. Up to 12 promoters can be independently controlled by the addition of up to 12 small-molecule inducers to the growth media.

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 Marionette Sensor Collection was a gift from Christopher Voigt (Addgene Kit #1000000137).”

For your Reference section:

Escherichia coli “Marionette” strains with 12 highly optimized small-molecule sensors. Meyer AJ, Segall-Shapiro TH, Glassey E, Zhang J, and Voigt CA. Nat Chem Biol. 2019; doi: 10.1038/s41589-018-0168-3. PubMed PMID 30478458.

Marionette Sensor Collection - #1000000137

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

Kanamycin
Low chloramphenicol (5 μg/ml)

Inventory

Well Plasmid Resistance
A / 1 pAJM.711
Kanamycin
A / 2 pAJM.712
Kanamycin
A / 3 pAJM.713
Kanamycin
A / 4 pAJM.714
Kanamycin
A / 5 pAJM.715
Kanamycin
A / 6 pAJM.717
Kanamycin
A / 7 pAJM.716
Kanamycin
A / 8 pAJM.718
Kanamycin
A / 9 pAJM.719
Kanamycin
A / 10 pAJM.1459
Kanamycin
A / 11 pAJM.721
Kanamycin
A / 12 pAJM.944
Kanamycin
B / 1 pAJM.847
Kanamycin
B / 2 pAJM.657
Kanamycin
B / 3 pAJM.474
Kanamycin
B / 4 pAJM.773
Kanamycin
B / 5 pAJM.336
Kanamycin
B / 6 pAJM.011
Kanamycin
B / 7 pAJM.677
Kanamycin
B / 8 pAJM.683
Kanamycin
B / 9 pAJM.661
Kanamycin
B / 10 pAJM.690
Kanamycin
B / 11 pAJM.771
Kanamycin
B / 12 pAJM.1642
Kanamycin
C / 1 pAJM.884
Kanamycin
C / 2 pAJM.969
Kanamycin
C / 3 sAJM.1504
Low chloramphenicol (5 μg/ml)
C / 4 sAJM.1505
Low chloramphenicol (5 μg/ml)
C / 5 sAJM.1506
Low chloramphenicol (5 μg/ml)
Data calculated @ 2019-07-18

Kit Plate Map - #1000000137

Plate #1

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