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: ampicillin (50 μg/ ml)
Position L1 (Weber et al): 2F
Brick name: PAR-BSR-TRBCS2 (TU)
Brick type: Antibiotic Resistance TU
Brick function: Resistance to Blasticidin in Chlamy
Source of Brick: Assembly of pCM0-020, pCM0-120 and pCM0-115
Acceptor plasmid: pICH47742
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

pCM1-029 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 Justus Niemeyer, Michael Schroda lab, University of Kaiserslautern, July 2021

Name: PSADpro::roGFP2-Tsa2ΔCR::3xHA::RPL23-T
Host strain: TOP10
Selection in E. coli: ampicillin (100 μg/ ml)
Type: 2F
Original pl1: pICH47742
Function: H2O2 reporter-3xHA tag
Brick 1 – pL0: pCM0-016
Brick 2 – pL0: pMBS419
Brick 3 – pL0: pCM0-093
Brick 4 – pL0: pCM0-119
Fusion 5′ lvl2: GCAA
Fusion 3′ lvl2: ACTA

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/

From Justus Niemeyer, Michael Schroda lab, University of Kaiserslautern, July 2021

Name: ARpro::roGFP2-Tsa2ΔCR::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: ampicillin (100 μg/ ml)
Type: 2F
Original pl1: pICH47742
Function: H2O2 reporter-no tag
Brick 1 – pL0: pCM0-017
Brick 2 – pL0: pMBS419
Brick 3 – pL0: pCM0-101
Brick 4 – pL0: pCM0-119
Fusion 5′ lvl2: GCAA
Fusion 3′ lvl2: ACTA

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/

From Justus Niemeyer, Michael Schroda lab, University of Kaiserslautern, July 2021

Name: ARpro::roGFP2::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: ampicillin (100 μg/ ml)
Type: 2F
Original pl1: pICH47742
Function: roGFP2 unfused-no tag
Brick 1 – pL0: pCM0-017
Brick 2 – pL0: pMBS467
Brick 3 – pL0: pCM0-101
Brick 4 – pL0: pCM0-119
Fusion 5′ lvl2: GCAA
Fusion 3′ lvl2: ACTA

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/

From Justus Niemeyer, Michael Schroda lab, University of Kaiserslautern, July 2021

Name: ARpro::USPA::roGFP2-Tsa2ΔCR::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: ampicillin (100 μg/ ml)
Type: 2F
Original pl1: pICH47742
Function: H2O2 reporter-no tag targeted to the chloroplast (Stroma)
Brick 1 – pL0: pCM0-011
Brick 2 – pL0: pCM0-053
Brick 3 – pL0: pMBS419
Brick 4 – pL0: pCM0-101
Brick 5 – pL0: pCM0-119
Fusion 5′ lvl2: GCAA
Fusion 3′ lvl2: ACTA

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/

From Justus Niemeyer, Michael Schroda lab, University of Kaiserslautern, July 2021

Name: ARpro::HSP70B::roGFP2-Tsa2ΔCR::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: ampicillin (100 μg/ ml)
Type: 2F
Original pl1: pICH47742
Function: H2O2 reporter-no tag targeted to the chloroplast (Stroma)
Brick 1 – pL0: pCM0-011
Brick 2 – pL0: pMBS639
Brick 3 – pL0: pMBS419
Brick 4 – pL0: pCM0-101
Brick 5 – pL0: pCM0-119
Fusion 5′ lvl2: GCAA
Fusion 3′ lvl2: ACTA

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/

From Justus Niemeyer, Michael Schroda lab, University of Kaiserslautern, July 2021

Name: ARpro::CDJ1::roGFP2-Tsa2ΔCR::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: ampicillin (100 μg/ ml)
Type: 2F
Original pl1: pICH47742
Function: H2O2 reporter-no tag targeted to the chloroplast (Stroma)
Brick 1 – pL0: pCM0-011
Brick 2 – pL0: pMBS640
Brick 3 – pL0: pMBS419
Brick 4 – pL0: pCM0-101
Brick 5 – pL0: pCM0-119
Fusion 5′ lvl2: GCAA
Fusion 3′ lvl2: ACTA

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/

From Justus Niemeyer, Michael Schroda lab, University of Kaiserslautern, July 2021

Name: ARpro::PSAN::roGFP2-Tsa2ΔCR::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: ampicillin (100 μg/ ml)
Type: 2F
Original pl1: pICH47742
Function: H2O2 reporter-no tag targeted to the chloroplast (Lumen)
Brick 1 – pL0: pCM0-011
Brick 2 – pL0: pMBS298
Brick 3 – pL0: pMBS419
Brick 4 – pL0: pCM0-101
Brick 5 – pL0: pCM0-119
Fusion 5′ lvl2: GCAA
Fusion 3′ lvl2: ACTA

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/

From Justus Niemeyer, Michael Schroda lab, University of Kaiserslautern, July 2021

Name: ARpro::PSBO::roGFP2-Tsa2ΔCR::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: ampicillin (100 μg/ ml)
Type: 2F
Original pl1: pICH47742
Function: H2O2 reporter-no tag targeted to the chloroplast (Lumen)
Brick 1 – pL0: pCM0-011 Brick 2 – pL0: pMBS641
Brick 3 – pL0: pMBS419 
Brick 4 – pL0: pCM0-101 
Brick 5 – pL0: pCM0-119 
Fusion 5′ lvl2: GCAA
Fusion 3′ lvl2: ACTA

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/

From Justus Niemeyer, Michael Schroda lab, University of Kaiserslautern, July 2021

Name: ARpro::HSP70C::roGFP2-Tsa2ΔCR::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: ampicillin (100 μg/ ml)
Type: 2F
Original pl1: pICH47742
Function: H2O2 reporter-no tag targeted to the mitochondria
Brick 1 – pL0: pCM0-011
Brick 2 – pL0: pCM0-057
Brick 3 – pL0: pMBS419
Brick 4 – pL0: pCM0-101
Brick 5 – pL0: pCM0-119
Fusion 5′ lvl2: GCAA
Fusion 3′ lvl2: ACTA

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/

From Justus Niemeyer, Michael Schroda lab, University of Kaiserslautern, July 2021

Name: ARpro::BIP1::roGFP2-Tsa2ΔCR::ER-ret::RPL23-T
Host strain: TOP10
Selection in E. coli: ampicillin (100 μg/ ml)
Type: 2F
Original pl1: pICH47742
Function: H2O2 reporter-no tag targeted to the endoplasmic reticulum
Brick 1 – pL0: pCM0-011
Brick 2 – pL0: pCM0-056
Brick 3 – pL0: pMBS419
Brick 4 – pL0: pCM0-111
Brick 5 – pL0: pCM0-119
Fusion 5′ lvl2: GCAA
Fusion 3′ lvl2: ACTA

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/

From Justus Niemeyer, Michael Schroda lab, University of Kaiserslautern, July 2021

Name: ARpro::roGFP2-Tsa2ΔCR::SV40::RPL23-T
Host strain: TOP10
Selection in E. coli: ampicillin (100 μg/ ml)
Type: 2F
Original pl1: pICH47742
Function: H2O2 reporter-no tag targeted to the nucleus
Brick 1 – pL0: pCM0-017
Brick 2 – pL0: pMBS419
Brick 3 – pL0: pCM0-109
Brick 4 – pL0: pCM0-119
Fusion 5′ lvl2: GCAA
Fusion 3′ lvl2: ACTA

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/

From Justus Niemeyer, Michael Schroda lab, University of Kaiserslautern, July 2021

Name: ARpro::roGFP2-Orp1::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: ampicillin (100 μg/ ml)
Type: 2F
Original pl1: pICH47742
Function: H2O2 reporter-no tag
Brick 1 – pL0: pCM0-017
Brick 2 – pL0: pMBS418
Brick 3 – pL0: pCM0-101
Brick 4 – pL0: pCM0-119
Fusion 5′ lvl2: GCAA
Fusion 3′ lvl2: ACTA

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/