MoClo Level 0 Parts

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: mVenus (2xi1) (CDS)
Brick type: Reporter gene (CDS)
Brick function: mVenus 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: https://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: DH5a
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B4
Brick name: Clover (2xi1) (CDS)
Brick type: Reporter gene (CDS)
Brick function: Clover 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: https://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: DH5a
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B4
Brick name: mRuby2 (2xi1) (CDS)
Brick type: Reporter gene (CDS)
Brick function: mRuby2 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: https://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: DH5a
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B3-B5
Brick name: aada (2xi1) (SpecR, CDS)
Brick type: Antibiotic Resistance gene (CDS)
Brick function: aadA confering resistance to Spectinomycine in Chlamy, codon optimized for Chlamy, including 2 copies of the rbcS2i1
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pICH41308
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: https://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: DH5a
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B4
Brick name: aada (2xi1) (SpecR, CDS)
Brick type: Antibiotic Resistance gene (CDS)
Brick function: aadA confering resistance to Spectinomycine in Chlamy, codon optimized for Chlamy, including 2 copies of the rbcS2i1
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: https://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: DH5a
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): A1-B1
Brick name: PSAD_mut-SspI (Pro + 5’UTR)
Brick type: promoter + 5’UTR
Brick function: PSAD promoter with mutated SspI site for L2 usage
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pAGM1251
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: https://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: DH5a
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B5
Brick name: 8xHis-Tag  (i2) (C-ter, CDS)
Brick type: Immuno- and purification tag
Brick function: Add a 8xHis 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: https://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: DH5a
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B5
Brick name: HA-Tag  (i2) (C-ter, CDS)
Brick type: Immuno- and purification tag
Brick function: Add a HA 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: https://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: DH5a
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B5
Brick name: FLAG-Tag  (i2) (C-ter, CDS)
Brick type: Immuno- and purification tag
Brick function: Add a FLAG 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: https://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 Thomas Baier, Olaf Kruse lab, Bielefeld University, August 2021

Host strain: DH5α
Selection in E. coli: spectinomycin (50 μg/ ml)
Position L0 (Patron et al): A1_B2
Brick name: PAßSAP(i) (Pro + 5’UTR)
Brick type: promoter + 5’UTR
Brick function: Synthetic HSP70A/ßtub promoter, which drives 4xstronger expression compared to pAR when transgene is expressed to the cytosol
Source of Brick: Einhaus et al. 2021
Acceptor plasmid: pICH41295
Designer / Builder: A. Einhaus and T. Baier

Sequence

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: https://www.chlamycollection.org/product/moclo-toolkit/

 

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): B3-B5
Brick name: NAT (NTCR, CDS)
Brick type: Antibiotic Resistance gene (CDS)
Brick function: Nourseothricin N-acetyl transferase (Yang et al. 2019) confering resistance to Noruseothricin in Chlamy, codon optimized for Chlamy
Source of Brick: Einhaus et al. 2022
Acceptor plasmid: pICH41308
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: https://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: DH5a
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al): B4
Brick name: shBle (i1) (shBle, CDS)
Brick type: Antibiotic Resistance gene (CDS)
Brick function: Bleomycine resistance gene, codon optimized for Chlamy, including RBCS2 intron 1, in B4 for translational fusions
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: https://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 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): A1-A3
Brick name: PPSAD (Pro)
Brick type: promoter
Brick function: Add PSAD promoter only (without 5’UTR)
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pICH41233
Designer / Builder: F. Navarro

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: https://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 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): A1-A3
Brick name: PAR (Pro)
Brick type: promoter
Brick function: Add HSP70A-RBSC2 tandem promoter
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pICH41233
Designer / Builder: F. Navarro

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: https://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 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): A1-A3
Brick name: PNIT1 (Pro)
Brick type: promoter
Brick function: Add NIT1 promoter
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pICH41233
Designer / Builder: F. Navarro

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: https://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 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): A1-A3
Brick name: PMETE (Pro)
Brick type: promoter
Brick function: METE promoter for inducible regulation of transgene expression with B12
Source of Brick: Cre03.g180750 (Helliwell et al, 2014)
Acceptor plasmid: pICH41233
Designer / Builder: A. Gorchs

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: https://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