MoClo Toolkit

The entire collection of plasmids, distributed as live cultures on two plates in a 96-well format, will be priced at $400 for academic/non-profit users and $800 for commercial/industry. Individual plasmids from the kit can also be ordered as bacterial cultures on agar plugs for standard plasmid pricing. You may view the individual list of plasmids in this collection here: https://www.chlamycollection.org/products/moclo-toolkit/. 

From Crozet et al. (2018): “… we have developed a Modular Cloning toolkit for the green microalga Chlamydomonas reinhardtii. It is based on Golden Gate cloning with standard syntax, and comprises 119 openly distributed genetic parts, most of which have been functionally validated in several strains. It contains promoters, UTRs, terminators, tags, reporters, antibiotic resistance genes, and introns cloned in various positions to allow maximum modularity. The toolkit enables rapid building of engineered cells for both fundamental research and algal biotechnology. This work will make Chlamydomonas the next chassis for sustainable synthetic biology.”

MoClo Kit Sequences

MoClo Kit Documents

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

We are sorry but we cannot ship the MoClo Toolkit to China because of long shipping and customs delays. Individual clones can still be shipped however. 

From Pablo Leivar, IQS School of Engineering-Barcelona Spain, December 2024

Host strain: DH5α
Selection in E. coli: Chloramphenicol (17µg/ml)
Position L0 (Patron et al): A1-B1
Brick name: PrbcL
Brick type: Promoter + 5’UTR
Brick function: Add rbcL promoter, 5’UTR and first 27 rbcL gene aminoacids
Source of Brick: Melero-Cobo et al., 2025
Acceptor plasmid: Twist vector_High copy, ChloroR
Designer / Builder: Melero-Cobo & Gallemí

Sequence

Melero-Cobo X, Gallemí M, Carnicer M, Monte E, Planas A, Leivar P. MoCloro: an extension of the Chlamydomonas reinhardtii modular cloning toolkit for microalgal chloroplast engineering. Physiol Plant. 2025 Jan-Feb;177(1):e70088. doi: 10.1111/ppl.70088. PMID: 39871102; PMCID: PMC11772913.

From Pablo Leivar, IQS School of Engineering-Barcelona Spain, December 2024

Host strain: DH5α
Selection in E. coli: Chloramphenicol (17µg/ml)
Position L0 (Patron et al): B6-C1
Brick name: TpsbA·T7
Brick type: 3’UTR + Terminator
Brick function: Add psbA terminator fused to T7 terminator
Source of Brick: Melero-Cobo et al., 2025
Acceptor plasmid: Twist vector_High copy, ChloroR
Designer / Builder: Melero-Cobo & Gallemí

Sequence

Melero-Cobo X, Gallemí M, Carnicer M, Monte E, Planas A, Leivar P. MoCloro: an extension of the Chlamydomonas reinhardtii modular cloning toolkit for microalgal chloroplast engineering. Physiol Plant. 2025 Jan-Feb;177(1):e70088. doi: 10.1111/ppl.70088. PMID: 39871102; PMCID: PMC11772913.

From Pablo Leivar, IQS School of Engineering-Barcelona Spain, December 2024

Host strain: DH5α
Selection in E. coli: Chloramphenicol (17µg/ml)
Position L0 (Patron et al): B6-C1
Brick name: TatpB
Brick type: 3’UTR + Terminator
Brick function: Add atpB terminator
Source of Brick: Melero-Cobo et al., 2025
Acceptor plasmid: Twist vector_High copy, ChloroR
Designer / Builder: Melero-Cobo & Gallemí

Sequence

Melero-Cobo X, Gallemí M, Carnicer M, Monte E, Planas A, Leivar P. MoCloro: an extension of the Chlamydomonas reinhardtii modular cloning toolkit for microalgal chloroplast engineering. Physiol Plant. 2025 Jan-Feb;177(1):e70088. doi: 10.1111/ppl.70088. PMID: 39871102; PMCID: PMC11772913.

From Pablo Leivar, IQS School of Engineering-Barcelona Spain, December 2024

Host strain: DH5α
Selection in E. coli: Chloramphenicol (17µg/ml)
Position L0 (Patron et al): B6-C1
Brick name: TpsaA
Brick type: 3’UTR + Terminator
Brick function: Add psaA terminator
Source of Brick: Melero-Cobo et al., 2025
Acceptor plasmid: Twist vector_High copy, ChloroR
Designer / Builder: Melero-Cobo & Gallemí

Sequence

Melero-Cobo X, Gallemí M, Carnicer M, Monte E, Planas A, Leivar P. MoCloro: an extension of the Chlamydomonas reinhardtii modular cloning toolkit for microalgal chloroplast engineering. Physiol Plant. 2025 Jan-Feb;177(1):e70088. doi: 10.1111/ppl.70088. PMID: 39871102; PMCID: PMC11772913.

From Pablo Leivar, IQS School of Engineering-Barcelona Spain, December 2024

Host strain: DH5α
Selection in E. coli: Chloramphenicol (17µg/ml)
Position L0 (Patron et al): A1-B1
Brick name: PpsbD
Brick type: Promoter + 5’UTR
Brick function: Add psbD promoter and 5’UTR
Source of Brick: Melero-Cobo et al., 2025
Acceptor plasmid: Twist vector_High copy, ChloroR
Designer / Builder: Melero-Cobo & Gallemí

Sequence

Melero-Cobo X, Gallemí M, Carnicer M, Monte E, Planas A, Leivar P. MoCloro: an extension of the Chlamydomonas reinhardtii modular cloning toolkit for microalgal chloroplast engineering. Physiol Plant. 2025 Jan-Feb;177(1):e70088. doi: 10.1111/ppl.70088. PMID: 39871102; PMCID: PMC11772913.

From Pablo Leivar, IQS School of Engineering-Barcelona Spain, December 2024

Host strain: DH5α
Selection in E. coli: Chloramphenicol (17µg/ml)
Position L0 (Patron et al): A1-B1
Brick name: P16SrRNA
Brick type: Promoter + 5’UTR
Brick function: Add 16SrRNA promoter and rbcL 5’UTR
Source of Brick: Melero-Cobo et al., 2025
Acceptor plasmid: Twist vector_High copy, ChloroR
Designer / Builder: Melero-Cobo & Gallemí

Sequence

Melero-Cobo X, Gallemí M, Carnicer M, Monte E, Planas A, Leivar P. MoCloro: an extension of the Chlamydomonas reinhardtii modular cloning toolkit for microalgal chloroplast engineering. Physiol Plant. 2025 Jan-Feb;177(1):e70088. doi: 10.1111/ppl.70088. PMID: 39871102; PMCID: PMC11772913.

From Pablo Leivar, IQS School of Engineering-Barcelona Spain, December 2024

Host strain: DH5α
Selection in E. coli: Chloramphenicol (17µg/ml)
Position L0 (Patron et al): B6-C1
Brick name: TsrbcL
Brick type: 3’UTR + Terminator
Brick function: Add a short version of the rbcL terminator
Source of Brick: Melero-Cobo et al., 2025
Acceptor plasmid: Twist vector_High copy, ChloroR
Designer / Builder: Melero-Cobo & Gallemí

Sequence

Melero-Cobo X, Gallemí M, Carnicer M, Monte E, Planas A, Leivar P. MoCloro: an extension of the Chlamydomonas reinhardtii modular cloning toolkit for microalgal chloroplast engineering. Physiol Plant. 2025 Jan-Feb;177(1):e70088. doi: 10.1111/ppl.70088. PMID: 39871102; PMCID: PMC11772913.

From Pablo Leivar, IQS School of Engineering-Barcelona Spain, December 2024

Host strain: DH5α
Selection in E. coli: Chloramphenicol (17µg/ml)
Position L0 (Patron et al): A1-B1
Brick name: PatpB
Brick type: Promoter + 5’UTR
Brick function: Add atpB promoter and 5’UTR
Source of Brick: Melero-Cobo et al., 2025
Acceptor plasmid: Twist vector_High copy, ChloroR
Designer / Builder: Melero-Cobo & Gallemí

Sequence

Melero-Cobo X, Gallemí M, Carnicer M, Monte E, Planas A, Leivar P. MoCloro: an extension of the Chlamydomonas reinhardtii modular cloning toolkit for microalgal chloroplast engineering. Physiol Plant. 2025 Jan-Feb;177(1):e70088. doi: 10.1111/ppl.70088. PMID: 39871102; PMCID: PMC11772913.

From Pablo Leivar, IQS School of Engineering-Barcelona Spain, December 2024

Host strain: DH5α
Selection in E. coli: Chloramphenicol (17µg/ml)
Position L0 (Patron et al): A1-B1
Brick name: PpsaA
Brick type: Promoter + 5’UTR
Brick function: Add psaA promoter and 5’UTR
Source of Brick: Melero-Cobo et al., 2025
Acceptor plasmid: Twist vector_High copy, ChloroR
Designer / Builder: Melero-Cobo & Gallemí

Sequence

Melero-Cobo X, Gallemí M, Carnicer M, Monte E, Planas A, Leivar P. MoCloro: an extension of the Chlamydomonas reinhardtii modular cloning toolkit for microalgal chloroplast engineering. Physiol Plant. 2025 Jan-Feb;177(1):e70088. doi: 10.1111/ppl.70088. PMID: 39871102; PMCID: PMC11772913.

From Pablo Leivar, IQS School of Engineering-Barcelona Spain, December 2024

Host strain: DH5α
Selection in E. coli: Chloramphenicol (17µg/ml)
Position L0 (Patron et al): B2-B5
Brick name: CDS uidA
Brick type: Reporter gene (CDS)
Brick function: Add uidA reporter gene sequence from E. coli
Source of Brick: Melero-Cobo et al., 2025
Acceptor plasmid: Twist vector_High copy, ChloroR
Designer / Builder: Melero-Cobo & Gallemí

Sequence

Melero-Cobo X, Gallemí M, Carnicer M, Monte E, Planas A, Leivar P. MoCloro: an extension of the Chlamydomonas reinhardtii modular cloning toolkit for microalgal chloroplast engineering. Physiol Plant. 2025 Jan-Feb;177(1):e70088. doi: 10.1111/ppl.70088. PMID: 39871102; PMCID: PMC11772913.

From Pablo Leivar, IQS School of Engineering-Barcelona Spain, December 2024

TU short name: PatpA·aadA·TrbcL
TU full name: pL1-2F-atpApro::aadA::TrbcL
Host strain: DH5α
Selection in E. coli: Ampicillin (100µg/ml)
Type: 2F
Original pl1: pICH47742
Function: Resistance in Chlamy chloroplast to Spectinomycin
Source of the module: Melero-Cobo et al., 2025
Designer/ Builder: Melero-Cobo & Gallemí
Fusion 5′ lvl2: GCAA
Fusion 3′ lvl2: ACTA
Comment: This module was not obtained by Golden Gate. Refer to Melero-Cobo et al. for further details.

Sequence

Melero-Cobo X, Gallemí M, Carnicer M, Monte E, Planas A, Leivar P. MoCloro: an extension of the Chlamydomonas reinhardtii modular cloning toolkit for microalgal chloroplast engineering. Physiol Plant. 2025 Jan-Feb;177(1):e70088. doi: 10.1111/ppl.70088. PMID: 39871102; PMCID: PMC11772913.

From Pablo Leivar, IQS School of Engineering-Barcelona Spain, December 2024

TU short name: PatpA·aadA·TrbcL
TU full name: pL1-1F-atpApro::aadA::TrbcL
Host strain: DH5α
Selection in E. coli: Ampicillin (100µg/ml)
Type: 1F
Original pl1: pICH47732
Function: Resistance in Chlamy chloroplast to Spectinomycin
Source of the module: Melero-Cobo et al., 2025
Designer/ Builder: Melero-Cobo & Gallemí
Fusion 5′ lvl2: TGCC
Fusion 3′ lvl2: GCAA
Comment: This module was not obtained by Golden Gate. Refer to Melero-Cobo et al. for further details.

Sequence

Melero-Cobo X, Gallemí M, Carnicer M, Monte E, Planas A, Leivar P. MoCloro: an extension of the Chlamydomonas reinhardtii modular cloning toolkit for microalgal chloroplast engineering. Physiol Plant. 2025 Jan-Feb;177(1):e70088. doi: 10.1111/ppl.70088. PMID: 39871102; PMCID: PMC11772913.

From Pablo Leivar, IQS School of Engineering-Barcelona Spain, December 2024

TU short name: PatpA·aadA·TrbcL
TU full name: pL1-3F-atpApro::aadA::TrbcL
Host strain: DH5α
Selection in E. coli: Ampicillin (100µg/ml)
Type: 3F
Original pl1: pICH47751
Function: Resistance in Chlamy chloroplast to Spectinomycin
Source of the module: Melero-Cobo et al., 2025
Designer/ Builder: Melero-Cobo & Gallemí
Fusion 5′ lvl2: ACTA
Fusion 3′ lvl2: TTAC
Comment: This module was not obtained by Golden Gate. Refer to Melero-Cobo et al. for further details.

Sequence

Melero-Cobo X, Gallemí M, Carnicer M, Monte E, Planas A, Leivar P. MoCloro: an extension of the Chlamydomonas reinhardtii modular cloning toolkit for microalgal chloroplast engineering. Physiol Plant. 2025 Jan-Feb;177(1):e70088. doi: 10.1111/ppl.70088. PMID: 39871102; PMCID: PMC11772913.

From Pablo Leivar, IQS School of Engineering-Barcelona Spain, December 2024

TU short name: PatpA·aadA·TrbcL
TU full name: pL1-4F-atpApro::aadA::TrbcL
Host strain: DH5α
Selection in E. coli: Ampicillin (100µg/ml)
Type: 4F
Original pl1: pICH47761
Function: Resistance in Chlamy chloroplast to Spectinomycin
Source of the module: Melero-Cobo et al., 2025
Designer/ Builder: Melero-Cobo & Gallemí
Fusion 5′ lvl2: TTAC
Fusion 3′ lvl2: CAGA
Comment: This module was not obtained by Golden Gate. Refer to Melero-Cobo et al. for further details.

Sequence

Melero-Cobo X, Gallemí M, Carnicer M, Monte E, Planas A, Leivar P. MoCloro: an extension of the Chlamydomonas reinhardtii modular cloning toolkit for microalgal chloroplast engineering. Physiol Plant. 2025 Jan-Feb;177(1):e70088. doi: 10.1111/ppl.70088. PMID: 39871102; PMCID: PMC11772913.

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.