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

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

Name: PSADpro::aadA::PSAD-T:::: ARpro::HSP70C::roGFP2-Tsa2ΔCR::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: Kanamycin (50 μg/ ml)
Original pL2: pAGM4673
Function: H2O2 reporter-no tag targeted to the mitochondria
TU1: pCM1-01
TU2: pMBS647
End Linker: pICH41744-pL2-EL2

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

From Thomas Baier, Olaf Kruse lab, Bielefeld University, August 2021

Host strain: DH5α
Selection in E. coli: Streptinomycin (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: http://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 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: http://www.chlamycollection.org/product/moclo-toolkit/

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

Name: PSADpro::aadA::PSAD-T:::: ARpro::BIP1::roGFP2-Tsa2ΔCR::ER-ret::RPL23-T
Host strain: TOP10
Selection in E. coli: Kanamycin (50 μg/ ml)
Original pL2: pAGM4673
Function: H2O2 reporter-no tag targeted to the endoplasmic reticulum
TU1: pCM1-01
TU2: pMBS648
End Linker: pICH41744-pL2-EL2

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

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

Name: PSADpro::aadA::PSAD-T:::: ARpro::roGFP2::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: Kanamycin (50 μg/ ml)
Original pL2: pAGM4673
Function: roGFP2 unfused-no tag
TU1: pCM1-01
TU2: pMBS470
End Linker: pICH41744-pL2-EL2

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

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

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

Name: PSADpro::aadA::PSAD-T:::: ARpro::roGFP2-Tsa2ΔCR::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: Kanamycin (50 μg/ ml)
Original pL2: pAGM4673
Function: H2O2 reporter-no tag
TU1: pCM1-01
TU2: pMBS469
End Linker: pICH41744-pL2-EL2

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

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

Name: PSADpro::aadA::PSAD-T:::: ARpro::roGFP2-Orp1::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: Kanamycin (50 μg/ ml)
Original pL2: pAGM4673
Function: H2O2 reporter-no tag
TU1: pCM1-01
TU2: pMBS719
End Linker: pICH41744-pL2-EL2

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

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

Name: PSADpro::aadA::PSAD-T:::: PSADpro::roGFP2-Tsa2ΔCR::3xHA::RPL23-T
Host strain: TOP10
Selection in E. coli: Kanamycin (50 μg/ ml)
Original pL2: pAGM4673
Function: H2O2 reporter-3xHA tag
TU1: pCM1-01
TU2: pMBS425
End Linker: pICH41744-pL2-EL2

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

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

Host strain: TOP10
Selection in E. coli: spectinomycin (50 μg/ ml)
Position L0 (Patron et al): B2
Brick name: CTP HSP70B  (N-ter, CDS)
Brick type: Signal and Targeting peptides (CDS)
Brick function: Allows the targetting to the Chloroplast
Source of Brick: Cre06.g250100.t1.2
Acceptor plasmid: pAGM1276
Designer / Builder: J. Niemeyer

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

 


Niemeyer J, Scheuring D, Oestreicher J, Morgan B, Schroda M. Real-time Monitoring of Subcellular H2O2 Distribution in Chlamydomonas reinhardtii. Plant Cell. 2021 Jul 1:koab176. doi: 10.1093/plcell/koab176. Epub ahead of print. PMID: 34196712.

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

From Justus Niemeyer, Michael Schroda lab, University of Kaiserslautern, 7/12/21

Name: PSADpro::aadA::PSAD-T:::: ARpro::USPA::roGFP2-Tsa2ΔCR::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: Kanamycin (50 μg/ ml)
Original pL2: pAGM4673
Function: H2O2 reporter-no tag targeted to the chloroplast (Stroma)
TU1: pCM1-01
TU2: pMBS642
End Linker: pICH41744-pL2-EL2

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

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

Host strain: TOP10
Selection in E. coli: spectinomycin (50 μg/ ml)
Position L0 (Patron et al): B2
Brick name: CTP CDJ1  (N-ter, CDS)
Brick type: Signal and Targeting peptides (CDS)
Brick function: Allows the targetting to the Chloroplast
Source of Brick: Cre12.g507650 
Acceptor plasmid: pAGM1276
Designer / Builder: J. Niemeyer

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

 


Niemeyer J, Scheuring D, Oestreicher J, Morgan B, Schroda M. Real-time Monitoring of Subcellular H2O2 Distribution in Chlamydomonas reinhardtii. Plant Cell. 2021 Jul 1:koab176. doi: 10.1093/plcell/koab176. Epub ahead of print. PMID: 34196712.

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

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

Name: PSADpro::aadA::PSAD-T:::: ARpro::HSP70B::roGFP2-Tsa2ΔCR::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: Kanamycin (50 μg/ ml)
Original pL2: pAGM4673
Function: H2O2 reporter-no tag targeted to the chloroplast (Stroma)
TU1: pCM1-01
TU2: pMBS643
End Linker: pICH41744-pL2-EL2

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

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

Host strain: TOP10
Selection in E. coli: spectinomycin (50 μg/ ml)
Position L0 (Patron et al): B2
Brick name: CTP PSAN  (N-ter, CDS)
Brick type: Signal and Targeting peptides (CDS)
Brick function: Allows the targetting to the Chloroplast
Source of Brick: Cre02.g082500 
Acceptor plasmid: pAGM1276
Designer / Builder: J. Niemeyer

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

 


Niemeyer J, Scheuring D, Oestreicher J, Morgan B, Schroda M. Real-time Monitoring of Subcellular H2O2 Distribution in Chlamydomonas reinhardtii. Plant Cell. 2021 Jul 1:koab176. doi: 10.1093/plcell/koab176. Epub ahead of print. PMID: 34196712.

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

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

Name: PSADpro::aadA::PSAD-T:::: ARpro::CDJ1::roGFP2-Tsa2ΔCR::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: Kanamycin (50 μg/ ml)
Original pL2: pAGM4673
Function: H2O2 reporter-no tag targeted to the chloroplast (Stroma)
TU1: pCM1-01
TU2: pMBS644
End Linker: pICH41744-pL2-EL2

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

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

Host strain: TOP10
Selection in E. coli: spectinomycin (50 μg/ ml)
Position L0 (Patron et al): B2
Brick name: CTP PSBO  (N-ter, CDS)
Brick type: Signal and Targeting peptides (CDS)
Brick function: Allows the targetting to the Chloroplast
Source of Brick: Cre09.g396213 
Acceptor plasmid: pAGM1276
Designer / Builder: J. Niemeyer

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

 


Niemeyer J, Scheuring D, Oestreicher J, Morgan B, Schroda M. Real-time Monitoring of Subcellular H2O2 Distribution in Chlamydomonas reinhardtii. Plant Cell. 2021 Jul 1:koab176. doi: 10.1093/plcell/koab176. Epub ahead of print. PMID: 34196712.

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

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

Name: PSADpro::aadA::PSAD-T:::: ARpro::PSAN::roGFP2-Tsa2ΔCR::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: Kanamycin (50 μg/ ml)
Original pL2: pAGM4673
Function: H2O2 reporter-no tag targeted to the chloroplast (Lumen)
TU1: pCM1-01
TU2: pMBS645
End Linker: pICH41744-pL2-EL2

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

The entire collection of plasmids, distributed as frozen glycerol stocks 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

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

Host strain: TOP10
Selection in E. coli: spectinomycin (50 μg/ ml)
Position L0 (Patron et al): B3-B4
Brick name: roGFP2-Orp1 (CDS)
Brick type: Reporter gene (CDS)
Brick function: roGFP2-Orp1 in CDS without STOP and that can be fused in C-Ter
Source of Brick: Synthetized
Acceptor plasmid: pAGM1287
Designer / Builder: J. Niemeyer

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

 


Niemeyer J, Scheuring D, Oestreicher J, Morgan B, Schroda M. Real-time Monitoring of Subcellular H2O2 Distribution in Chlamydomonas reinhardtii. Plant Cell. 2021 Jul 1:koab176. doi: 10.1093/plcell/koab176. Epub ahead of print. PMID: 34196712.

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

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

Name: PSADpro::aadA::PSAD-T:::: ARpro::PSBO::roGFP2-Tsa2ΔCR::MStop::RPL23-T
Host strain: TOP10
Selection in E. coli: Kanamycin (50 μg/ ml)
Original pL2: pAGM4673
Function: H2O2 reporter-no tag targeted to the chloroplast (Lumen)
TU1: pCM1-01
TU2: pMBS646
End Linker: pICH41744-pL2-EL2

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

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

Host strain: TOP10
Selection in E. coli: spectinomycin (50 μg/ ml)
Position L0 (Patron et al): B3-B4
Brick name: roGFP2-Tsa2ΔCR (CDS)
Brick type: Reporter gene (CDS)
Brick function: roGFP2-Tsa2ΔCR in CDS without STOP and that can be fused in C-Ter
Source of Brick: Synthetized
Acceptor plasmid: pAGM1287
Designer / Builder: J. Niemeyer

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

 


Niemeyer J, Scheuring D, Oestreicher J, Morgan B, Schroda M. Real-time Monitoring of Subcellular H2O2 Distribution in Chlamydomonas reinhardtii. Plant Cell. 2021 Jul 1:koab176. doi: 10.1093/plcell/koab176. Epub ahead of print. PMID: 34196712.