MoClo Toolkit

From Pierre Crozet, Institute of Physico-Chemical Biology-Paris, December 2018

Plasmid used in the Modular Cloning Toolkit (MoClo) from Crozet et al. (2018).

Host strain: TOP10
Selection in E. coli: spectinomycin (50 μg/ ml)
Position L0 (Patron et al): B6-C1
Brick name: TRPL23 (i) (3´UTR + Ter)
Brick type: 3’UTR + Terminator
Brick function: 3’UTR (with intron) and Terminator for expression
Source of Brick: Cre04.g211800
Acceptor plasmid: pICH41276
Designer / Builder: B. Spaniol

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Crozet et al. (2018) Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii. ACS Synthetic Biology 7:2074–2086

From Félix de Carpentier, Institute of Physico-Chemical Biology-Paris, February 2020

Plasmid used in the Modular Cloning Toolkit (MoClo) from Crozet et al. (2018) and de Carpentier et al. (2020).

Host strain: NEB^® 10-beta
Selection in E. coli: spectinomycin (50 μg/ ml)
Position L0 (Patron et al): B3-B5
Brick name: BSR (CDS)
Brick type: Antibiotic Resistance gene (CDS)
Brick function: Resistance to Blasticidin in Chlamy
Source of Brick: Synthetized
Acceptor plasmid: pICH41308
Designer / Builder: F. de Carpentier and J. Le Peillet

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Crozet et al. (2018) Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii. ACS Synthetic Biology 7:2074–2086

de Carpentier et al. (2020) Blasticidin S deaminase: a new efficient selectable marker for Chlamydomonas reinhardtii. Frontiers in Plant Science 11:242

pCM0-120 sequence

de Carpentier F, Le Peillet J, Boisset ND, Crozet P, Lemaire SD, Danon A. Blasticidin S Deaminase: A New Efficient Selectable Marker for Chlamydomonas reinhardtii. Front Plant Sci. 2020 Mar 5;11:242. doi: 10.3389/fpls.2020.00242. PMID: 32211000; PMCID: PMC7066984.

From Alexander Einhaus and Dr. Thomas Baier, Bielefeld University-Germany, April 2022

Host strain: DH5a
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B6-C1
Brick name: FDX (3´UTR + Ter)
Brick type: 3’UTR + Terminator
Brick function: 3’UTR and Terminator for expression (Lopez-Paz et al. 2017)
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pICH41276
Designer / Builder: A. Einhaus and T. Baier

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Sequence

López-Paz C, Liu D, Geng S, Umen JG. Identification of Chlamydomonas reinhardtii endogenous genic flanking sequences for improved transgene expression. Plant J. 2017 Dec;92(6):1232-1244. doi: 10.1111/tpj.13731. Epub 2017 Nov 18. PMID: 28980350; PMCID: PMC5718938.

Crozet P, Navarro FJ, Willmund F, Mehrshahi P, Bakowski K, Lauersen KJ, Pérez-Pérez ME, Auroy P, Gorchs Rovira A, Sauret-Gueto S, Niemeyer J, Spaniol B, Theis J, Trösch R, Westrich LD, Vavitsas K, Baier T, Hübner W, de Carpentier F, Cassarini M, Danon A, Henri J, Marchand CH, de Mia M, Sarkissian K, Baulcombe DC, Peltier G, Crespo JL, Kruse O, Jensen PE, Schroda M, Smith AG, Lemaire SD. Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii. ACS Synth Biol. 2018 Sep 21;7(9):2074-2086. doi: 10.1021/acssynbio.8b00251. Epub 2018 Sep 5. PMID: 30165733.

Einhaus A, Baier T, Rosenstengel M, Freudenberg RA, Kruse O. Rational Promoter Engineering Enables Robust Terpene Production in Microalgae. ACS Synth Biol. 2021 Apr 16;10(4):847-856. doi: 10.1021/acssynbio.0c00632. Epub 2021 Mar 25. PMID: 33764741.

From Alexander Einhaus and Dr. Thomas Baier, Bielefeld University-Germany, April 2022

Host strain: DH5a
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B5
Brick name: StrepII (i2)  (C-ter, CDS)
Brick type: Immuno- and purification tag
Brick function: Add a StrepII tag (with RBCS2 intron 2) + STOP in C-terminal
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pAGM1301
Designer / Builder: A. Einhaus and T. Baier

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Sequence

Crozet P, Navarro FJ, Willmund F, Mehrshahi P, Bakowski K, Lauersen KJ, Pérez-Pérez ME, Auroy P, Gorchs Rovira A, Sauret-Gueto S, Niemeyer J, Spaniol B, Theis J, Trösch R, Westrich LD, Vavitsas K, Baier T, Hübner W, de Carpentier F, Cassarini M, Danon A, Henri J, Marchand CH, de Mia M, Sarkissian K, Baulcombe DC, Peltier G, Crespo JL, Kruse O, Jensen PE, Schroda M, Smith AG, Lemaire SD. Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii. ACS Synth Biol. 2018 Sep 21;7(9):2074-2086. doi: 10.1021/acssynbio.8b00251. Epub 2018 Sep 5. PMID: 30165733.

Einhaus A, Steube J, Freudenberg RA, Barczyk J, Baier T, Kruse O. Engineering a powerful green cell factory for robust photoautotrophic diterpenoid production. Metab Eng. 2022 Jun 16;73:82-90. doi: 10.1016/j.ymben.2022.06.002. Epub ahead of print. PMID: 35717002.

From Alexander Einhaus and Dr. Thomas Baier, Bielefeld University-Germany, April 2022

Host strain: TOP10
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B3
Brick name: MvELS (CDS_B3)
Brick type: Gene of interest (CDS)
Sequence reference: Uniprot: A0A075FBG7
Brick function: functional diterpene synthase from Marrubium vulgare, sequence optimized according to Baier et al. 2018
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pICH41258
Designer / Builder: A. Einhaus and T. Baier

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Sequence

Baier T, Wichmann J, Kruse O, Lauersen KJ. Intron-containing algal transgenes mediate efficient recombinant gene expression in the green microalga Chlamydomonas reinhardtii. Nucleic Acids Res. 2018 Jul 27;46(13):6909-6919. doi: 10.1093/nar/gky532. PMID: 30053227; PMCID: PMC6061784.

Einhaus A, Steube J, Freudenberg RA, Barczyk J, Baier T, Kruse O. Engineering a powerful green cell factory for robust photoautotrophic diterpenoid production. Metab Eng. 2022 Jun 16;73:82-90. doi: 10.1016/j.ymben.2022.06.002. Epub ahead of print. PMID: 35717002.

From Alexander Einhaus and Dr. Thomas Baier, Bielefeld University-Germany, April 2022

Host strain: TOP10
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B3
Brick name: SsSCS (CDS_B3)
Brick type: Gene of interest (CDS)
Sequence reference: Uniprot: K4HYB0
Brick function: functional diterpene synthase from Salvia sclarea, sequence optimized according to Baier et al. 2018
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pICH41258
Designer / Builder: A. Einhaus and T. Baier

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Sequence

Baier T, Wichmann J, Kruse O, Lauersen KJ. Intron-containing algal transgenes mediate efficient recombinant gene expression in the green microalga Chlamydomonas reinhardtii. Nucleic Acids Res. 2018 Jul 27;46(13):6909-6919. doi: 10.1093/nar/gky532. PMID: 30053227; PMCID: PMC6061784.

Einhaus A, Steube J, Freudenberg RA, Barczyk J, Baier T, Kruse O. Engineering a powerful green cell factory for robust photoautotrophic diterpenoid production. Metab Eng. 2022 Jun 16;73:82-90. doi: 10.1016/j.ymben.2022.06.002. Epub ahead of print. PMID: 35717002.

From Alexander Einhaus and Dr. Thomas Baier, Bielefeld University-Germany, April 2022

Host strain: TOP10
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B3
Brick name: SsLPPS (CDS_B3)
Brick type: Gene of interest (CDS)
Sequence reference: Uniprot: K4HZA9
Brick function: functional diterpene synthase from Salvia sclarea, sequence optimized according to Baier et al. 2018
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pICH41258
Designer / Builder: A. Einhaus and T. Baier

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Sequence

Baier T, Wichmann J, Kruse O, Lauersen KJ. Intron-containing algal transgenes mediate efficient recombinant gene expression in the green microalga Chlamydomonas reinhardtii. Nucleic Acids Res. 2018 Jul 27;46(13):6909-6919. doi: 10.1093/nar/gky532. PMID: 30053227; PMCID: PMC6061784.

Einhaus A, Steube J, Freudenberg RA, Barczyk J, Baier T, Kruse O. Engineering a powerful green cell factory for robust photoautotrophic diterpenoid production. Metab Eng. 2022 Jun 16;73:82-90. doi: 10.1016/j.ymben.2022.06.002. Epub ahead of print. PMID: 35717002.

From Alexander Einhaus and Dr. Thomas Baier, Bielefeld University-Germany, April 2022

Host strain: TOP10
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B3
Brick name: CfTPS1 (CDS_B3)
Brick type: Gene of interest (CDS)
Sequence reference: Uniprot: X5A4D6
Brick function: functional diterpene synthase from Coleus forskohlii, sequence optimized according to Baier et al. 2018
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pICH41258
Designer / Builder: A. Einhaus and T. Baier

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Sequence

Baier T, Wichmann J, Kruse O, Lauersen KJ. Intron-containing algal transgenes mediate efficient recombinant gene expression in the green microalga Chlamydomonas reinhardtii. Nucleic Acids Res. 2018 Jul 27;46(13):6909-6919. doi: 10.1093/nar/gky532. PMID: 30053227; PMCID: PMC6061784.

Einhaus A, Steube J, Freudenberg RA, Barczyk J, Baier T, Kruse O. Engineering a powerful green cell factory for robust photoautotrophic diterpenoid production. Metab Eng. 2022 Jun 16;73:82-90. doi: 10.1016/j.ymben.2022.06.002. Epub ahead of print. PMID: 35717002.

From Alexander Einhaus and Dr. Thomas Baier, Bielefeld University-Germany, April 2022

Host strain: TOP10
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B3
Brick name: TwTPS14 (CDS_B3)
Brick type: Gene of interest (CDS)
Sequence reference: Uniprot: A0A0M5L832
Brick function: functional diterpene synthase from Tripterygium wilfordii, sequence optimized according to Baier et al. 2018
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pICH41258
Designer / Builder: A. Einhaus and T. Baier

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Sequence

Baier T, Wichmann J, Kruse O, Lauersen KJ. Intron-containing algal transgenes mediate efficient recombinant gene expression in the green microalga Chlamydomonas reinhardtii. Nucleic Acids Res. 2018 Jul 27;46(13):6909-6919. doi: 10.1093/nar/gky532. PMID: 30053227; PMCID: PMC6061784.

Einhaus A, Steube J, Freudenberg RA, Barczyk J, Baier T, Kruse O. Engineering a powerful green cell factory for robust photoautotrophic diterpenoid production. Metab Eng. 2022 Jun 16;73:82-90. doi: 10.1016/j.ymben.2022.06.002. Epub ahead of print. PMID: 35717002.

From Alexander Einhaus and Dr. Thomas Baier, Bielefeld University-Germany, April 2022

Host strain: TOP10
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B3
Brick name: CfTPS3 (CDS_B3)
Brick type: Gene of interest (CDS)
Sequence reference: Uniprot: X5A2Z7
Brick function: functional diterpene synthase from Coleus forskohlii, sequence optimized according to Baier et al. 2018
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pICH41258
Designer / Builder: A. Einhaus and T. Baier

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Sequence

Baier T, Wichmann J, Kruse O, Lauersen KJ. Intron-containing algal transgenes mediate efficient recombinant gene expression in the green microalga Chlamydomonas reinhardtii. Nucleic Acids Res. 2018 Jul 27;46(13):6909-6919. doi: 10.1093/nar/gky532. PMID: 30053227; PMCID: PMC6061784.

Einhaus A, Steube J, Freudenberg RA, Barczyk J, Baier T, Kruse O. Engineering a powerful green cell factory for robust photoautotrophic diterpenoid production. Metab Eng. 2022 Jun 16;73:82-90. doi: 10.1016/j.ymben.2022.06.002. Epub ahead of print. PMID: 35717002.

From Alexander Einhaus and Dr. Thomas Baier, Bielefeld University-Germany, April 2022

Host strain: DH5a
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B4
Brick name: mCerulean3 (2xi1) (CDS)
Brick type: Reporter gene (CDS)
Brick function: mCerulean3 reporter (with 2 copies of the RBCS2 intron 1) + GSG-Linkers in B4
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pAGM1299
Designer / Builder: A. Einhaus and T. Baier

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Sequence

Crozet P, Navarro FJ, Willmund F, Mehrshahi P, Bakowski K, Lauersen KJ, Pérez-Pérez ME, Auroy P, Gorchs Rovira A, Sauret-Gueto S, Niemeyer J, Spaniol B, Theis J, Trösch R, Westrich LD, Vavitsas K, Baier T, Hübner W, de Carpentier F, Cassarini M, Danon A, Henri J, Marchand CH, de Mia M, Sarkissian K, Baulcombe DC, Peltier G, Crespo JL, Kruse O, Jensen PE, Schroda M, Smith AG, Lemaire SD. Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii. ACS Synth Biol. 2018 Sep 21;7(9):2074-2086. doi: 10.1021/acssynbio.8b00251. Epub 2018 Sep 5. PMID: 30165733.

Einhaus A, Steube J, Freudenberg RA, Barczyk J, Baier T, Kruse O. Engineering a powerful green cell factory for robust photoautotrophic diterpenoid production. Metab Eng. 2022 Jun 16;73:82-90. doi: 10.1016/j.ymben.2022.06.002. Epub ahead of print. PMID: 35717002.

From Alexander Einhaus and Dr. Thomas Baier, Bielefeld University-Germany, April 2022

Host strain: TOP10
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B3
Brick name: SpDXS_K284N (CDS_B3)
Brick type: Gene of interest (CDS)
Sequence reference: Uniprot: A0A4V1ES21
Brick function: functional diterpene synthase from salvia pomifera, mutated K284N, sequence optimized according to Baier et al. 2018
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pICH41258
Designer / Builder: A. Einhaus and T. Baier

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Sequence

Baier T, Wichmann J, Kruse O, Lauersen KJ. Intron-containing algal transgenes mediate efficient recombinant gene expression in the green microalga Chlamydomonas reinhardtii. Nucleic Acids Res. 2018 Jul 27;46(13):6909-6919. doi: 10.1093/nar/gky532. PMID: 30053227; PMCID: PMC6061784.

Einhaus A, Steube J, Freudenberg RA, Barczyk J, Baier T, Kruse O. Engineering a powerful green cell factory for robust photoautotrophic diterpenoid production. Metab Eng. 2022 Jun 16;73:82-90. doi: 10.1016/j.ymben.2022.06.002. Epub ahead of print. PMID: 35717002.

From Alexander Einhaus and Dr. Thomas Baier, Bielefeld University-Germany, April 2022

Host strain: TOP10
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B5
Brick name: CrGGPPS (CDS_B5)
Brick type: Gene of interest (CDS)
Sequence reference: Uniprot: A8JHU6
Brick function: functional geranylgeranyl pyrophosphate synthase from Chlamydomonas reinhardtii, sequence optimized according to Baier et al. 2018
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pAGM1301
Designer / Builder: A. Einhaus and T. Baier

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Sequence

Baier T, Wichmann J, Kruse O, Lauersen KJ. Intron-containing algal transgenes mediate efficient recombinant gene expression in the green microalga Chlamydomonas reinhardtii. Nucleic Acids Res. 2018 Jul 27;46(13):6909-6919. doi: 10.1093/nar/gky532. PMID: 30053227; PMCID: PMC6061784.

Einhaus A, Steube J, Freudenberg RA, Barczyk J, Baier T, Kruse O. Engineering a powerful green cell factory for robust photoautotrophic diterpenoid production. Metab Eng. 2022 Jun 16;73:82-90. doi: 10.1016/j.ymben.2022.06.002. Epub ahead of print. PMID: 35717002.

From Alexander Einhaus and Dr. Thomas Baier, Bielefeld University-Germany, April 2022

Host strain: TOP10
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B3
Brick name: HlDXR (CDS_B3)
Brick type: Gene of interest (CDS)
Sequence reference: Uniprot: H2DQ70
Brick function: functional 1-deoxy-d-xylulose 5-phosphate reductoisomerase from Haematococcus lacustris, sequence optimized according to Baier et al. 2018
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pICH41258
Designer / Builder: A. Einhaus and T. Baier

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Sequence

Baier T, Wichmann J, Kruse O, Lauersen KJ. Intron-containing algal transgenes mediate efficient recombinant gene expression in the green microalga Chlamydomonas reinhardtii. Nucleic Acids Res. 2018 Jul 27;46(13):6909-6919. doi: 10.1093/nar/gky532. PMID: 30053227; PMCID: PMC6061784.

Einhaus A, Steube J, Freudenberg RA, Barczyk J, Baier T, Kruse O. Engineering a powerful green cell factory for robust photoautotrophic diterpenoid production. Metab Eng. 2022 Jun 16;73:82-90. doi: 10.1016/j.ymben.2022.06.002. Epub ahead of print. PMID: 35717002.

From Alexander Einhaus and Dr. Thomas Baier, Bielefeld University-Germany, April 2022

Host strain: TOP10
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B3
Brick name: CfTPS2 (CDS_B3)
Brick type: Gene of interest (CDS)
Sequence reference: Uniprot: X4ZWN5
Brick function: functional diterpene synthase from Coleus forskohlii, sequence optimized according to Baier et al. 2018
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pICH41258
Designer / Builder: A. Einhaus and T. Baier

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Sequence

Baier T, Wichmann J, Kruse O, Lauersen KJ. Intron-containing algal transgenes mediate efficient recombinant gene expression in the green microalga Chlamydomonas reinhardtii. Nucleic Acids Res. 2018 Jul 27;46(13):6909-6919. doi: 10.1093/nar/gky532. PMID: 30053227; PMCID: PMC6061784.

Einhaus A, Steube J, Freudenberg RA, Barczyk J, Baier T, Kruse O. Engineering a powerful green cell factory for robust photoautotrophic diterpenoid production. Metab Eng. 2022 Jun 16;73:82-90. doi: 10.1016/j.ymben.2022.06.002. Epub ahead of print. PMID: 35717002.

From Alexander Einhaus and Dr. Thomas Baier, Bielefeld University-Germany, April 2022

Host strain: TOP10
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B3
Brick name: ScIDI (CDS_B3)
Brick type: Gene of interest (CDS)
Sequence reference: Uniprot: P15496
Brick function: functional isopentenyl diphosphate isomerase from Saccharomyces cerevisiae, sequence optimized according to Baier et al. 2018
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pICH41258
Designer / Builder: A. Einhaus and T. Baier

The entire collection of MoClo Toolkit plasmids, distributed as frozen glycerol stocks in a 96-well format, can also be ordered for $400 for academic/non-profit users and $800 for commercial/industry here: http://www.chlamycollection.org/product/moclo-toolkit/

Sequence

Baier T, Wichmann J, Kruse O, Lauersen KJ. Intron-containing algal transgenes mediate efficient recombinant gene expression in the green microalga Chlamydomonas reinhardtii. Nucleic Acids Res. 2018 Jul 27;46(13):6909-6919. doi: 10.1093/nar/gky532. PMID: 30053227; PMCID: PMC6061784.

Einhaus A, Steube J, Freudenberg RA, Barczyk J, Baier T, Kruse O. Engineering a powerful green cell factory for robust photoautotrophic diterpenoid production. Metab Eng. 2022 Jun 16;73:82-90. doi: 10.1016/j.ymben.2022.06.002. Epub ahead of print. PMID: 35717002.