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): B5
Brick name: Clover (i2)-Strep (C-ter, CDS)
Brick type: Reporter gene (CDS)
Brick function: Add a Clover (with RBCS2 intron 2) + StrepII tag in C-terminal
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pAGM1301
Designer / Builder: KJ. Lauersen

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): B5
Brick name: mCerulean3 (i2)-Strep (C-ter, CDS)
Brick type: Reporter gene (CDS)
Brick function: Add an mCerulean3 (with RBCS2 intron 2) + StrepII tag in C-terminal
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pAGM1301
Designer / Builder: KJ. Lauersen

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): B5
Brick name: gLuc (i2)-Strep (C-ter, CDS)
Brick type: Reporter gene (CDS)
Brick function: Add a gLuc (with RBCS2 intron 2) + StrepII tag in C-terminal
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pAGM1301
Designer / Builder: KJ. Lauersen

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): B5
Brick name: mCherry (i1) (C-ter, CDS)
Brick type: Reporter gene (CDS)
Brick function: red FP (RFP) with the intron 1 of RBCS2
Source of Brick: Nelson et al, 2007
Acceptor plasmid: pAGM1301
Designer / Builder: L. Westrich

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): B5
Brick name: NLS SV40 (C-ter, CDS)
Brick type: Signal and Targeting peptides (CDS)
Brick function: Allows the targetting to the nucleus
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pAGM1301
Designer / Builder: KJ. Lauersen

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): B5
Brick name: CrMSPTS1 (C-ter, CDS)
Brick type: Signal and Targeting peptides (CDS)
Brick function: Allows the targetting to the microbodies
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pAGM1301
Designer / Builder: KJ. Lauersen

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): B5
Brick name: ER ret (C-ter, CDS)
Brick type: Signal and Targeting peptides (CDS)
Brick function: Allows retention in Endoplasmic Reticulum
Source of Brick: pBR30 (Rasala et al, 2013)
Acceptor plasmid: pAGM1301
Designer / Builder: P. Auroy

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): B5
Brick name: mRuby2 (i2)-Strep (C-ter, CDS)
Brick type: Reporter gene (CDS)
Brick function: Add an mRuby2 (with RBCS2 intron 2) + StrepII tag in C-terminal
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pAGM1301
Designer / Builder: KJ. Lauersen

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): B5
Brick name: mVenus (i2)-Strep (C-ter, CDS)
Brick type: Reporter gene (CDS)
Brick function: Add an mVenus (with RBCS2 intron 2) + StrepII tag in C-terminal
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pAGM1301
Designer / Builder: KJ. Lauersen

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): B6-C1
Brick name: TPSAD (3´UTR + Ter)
Brick type: 3’UTR + Terminator
Brick function: 3’UTR and Terminator for expression
Source of Brick: Cre05.g238332
Acceptor plasmid: pICH41276
Designer / Builder: P. Crozet

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): B6-C1
Brick name: TRBCS2 (3´UTR + Ter)
Brick type: 3’UTR + Terminator
Brick function: 3’UTR and Terminator for expression
Source of Brick: Cre02.g120150
Acceptor plasmid: pICH41276
Designer / Builder: P. Crozet

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): B6-C1
Brick name: TRPS29 (3´UTR + Ter)
Brick type: 3’UTR + Terminator
Brick function: 3’UTR and Terminator for expression
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pICH41276
Designer / Builder: D. Gangl

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): B6-C1
Brick name: TCA1 (3´UTR + Ter)
Brick type: 3’UTR + Terminator
Brick function: 3’UTR and Terminator for expression
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pICH41276
Designer / Builder: P. Mehrshahi

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): B6-C1
Brick name: TβTUB2 (3’UTR + Ter)
Brick type: 3’UTR + Terminator
Brick function: 3’UTR and Terminator for strong expression
Source of Brick: Cre12.g549550
Acceptor plasmid: pICH41276
Designer / Builder: L. Westrich

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): 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: 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 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: 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

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.