MoClo Level 0 Parts

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): B3-B5
Brick name: amiRNA (SpeI internal site, CDS)
Brick type: amiRs (CDS)
Brick function: Add a miR1157 precursor domesticated sequence with SpeI sites for cloning amiRNAs as in pChlamiRNA3
Source of Brick: Synthetized (Molnar et al, 2007)
Acceptor plasmid: pICH41308
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): B3-B5
Brick name: Ble(i)GFP (CDS)
Brick type: Antibiotic Resistance + Reporter gene (CDS)
Brick function: Ble(i) GFP fusion protein provinding resitance to Bleomycin or Zeocin
Source of Brick: Fuhrmann et al., 1999
Acceptor plasmid: pAGM1287
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): B3-B5
Brick name: AphVII (HygroR, CDS)
Brick type: Antibiotic Resistance gene (CDS)
Brick function: Resistance to Hygromycine in Chlamy
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pICH41308
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): B3-B5
Brick name: AphVIII (ParoR, CDS)
Brick type: Antibiotic Resistance gene (CDS)
Brick function: Resistance to Paromomycine in Chlamy
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pICH41308
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): B3-B5
Brick name: CrnptII (KanR, CDS)
Brick type: Antibiotic Resistance gene (CDS)
Brick function: Gives Kanamycin resistance in Chlamydomonas
Source of Brick: pRMB27 (Barahimipour et al, 2015)
Acceptor plasmid: pICH41308
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): B3-B5
Brick name: aadA (SpecR, CDS)
Brick type: Antibiotic Resistance gene (CDS)
Brick function: Confers resistance to Spectinomycine in Chlamy
Source of Brick: aadA (Cerutti et al, 1997)
Acceptor plasmid: pICH41308
Designer / Builder: F. Willmund

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): B3-B5
Brick name: Bleocmycin (i) selection marker (BleoR, CDS)
Brick type: Antibiotic Resistance gene (CDS)
Brick function: provides resistance to Bleomycin or Zeocin (CDS with intron)
Source of Brick: Stevens et al, 1996
Acceptor plasmid: pICH41308
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): B3-B5
Brick name: NanoLuc (CDS)
Brick type: Reporter gene (CDS)
Brick function: NanoLuc for testing promoters & terminators combinations
Source of Brick: Synthetized (Hall et al, 2012)
Acceptor plasmid: pICH41308
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): B3-B5
Brick name: Clover(i2) Strep (CDS)
Brick type: Reporter gene (CDS)
Brick function: Add a Clover (with RBCS2 intron 2) + StrepII tag in CDS1
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pICH41308
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): B3-B5
Brick name: mCerulean3(i2) Strep (CDS)
Brick type: Reporter gene (CDS)
Brick function: Add an mCerulean3 (with RBCS2 intron 2) + StrepII tag in CDS1
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pICH41308
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): B3-B5
Brick name: gLuc(i2) Strep (CDS)
Brick type: Reporter gene (CDS)
Brick function: Add a gLuc (with RBCS2 intron 2) + StrepII tag in CDS1
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pICH41308
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): B3-B5
Brick name: mCherry(i1) (CDS)
Brick type: Reporter gene (CDS)
Brick function: mCherry2 (with RBCS2 intron1)
Source of Brick: Synthetized
Acceptor plasmid: pICH41308
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): B3-B5
Brick name: mRuby2(i2)-Strep (CDS)
Brick type: Reporter gene (CDS)
Brick function: Add an mRuby2 (with RBCS2 intron 2) + StrepII tag in CDS1
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pICH41308
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): B3-B5
Brick name: mVenus (i2)-Strep (CDS)
Brick type: Reporter gene (CDS)
Brick function: Add an mVenus (with RBCS2 intron 2) + StrepII tag in CDS1
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pICH41308
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): B4
Brick name: 2A B4
Brick type: 2A peptide (CDS)
Brick function: Allows expression of 2 proteines with same transcription (clivage during traduction)
Source of Brick: Synthetized from pchlamy 5 (Plucinak et al, 2015)
Acceptor plasmid: pAGM1299
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): B4
Brick name: SP cCA (N-ter, CDS)
Brick type: Signal and Targeting peptides (CDS)
Brick function: Allows secretion to the extracellular matrix
Source of Brick: Lauersen et al, 2015
Acceptor plasmid: pAGM1299
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