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


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 Anthony Berndt, Stephen Mayfield Lab, University of California, San Diego, November 2021

Vector: pUC19-based backbone
Host strain: DH5alpha
Origin: Gibson assembly using pUC19 backbone from synthesized gBlocks and elements of the pOpt vectors from the Kruse Lab.
Insert: Chloroplast directed SARS-CoV-2 Spike RBD fused to mClover fluorescent protein
Selection in E. coli: Ampicillin
Comment: Transformed into algae strain RMC1. pRMC1 map in GenBank format is also available in publication’s supplemental data.

Sequence


Berndt AJ, Smalley TN, Ren B, Simkovsky R, Badary A, Sproles AE, Fields FJ, Torres-Tiji Y, Heredia V, Mayfield SP. Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii. PLoS One. 2021 Nov 18;16(11):e0257089. doi: 10.1371/journal.pone.0257089. PMID: 34793485; PMCID: PMC8601568.

From Anthony Berndt, Stephen Mayfield Lab, University of California, San Diego, November 2021

Source: Anthony Berndt, Stephen Mayfield Lab, University of California, San Diego
Plasmid: pRMC2
Vector: pUC19-based backbone
Host strain: DH5alpha
Origin: Gibson assembly using pUC19 backbone from synthesized gBlocks and elements of the pOpt vectors from the Kruse Lab.
Insert: Secretion directed SARS-CoV-2 Spike RBD fused to mClover fluorescent protein.
Selection in E. coli: Ampicillin
Comment: Transformed in to algae strain RMC2. pRMC2 map in GenBank format is also available in publication’s supplemental data.

Sequence


Berndt AJ, Smalley TN, Ren B, Simkovsky R, Badary A, Sproles AE, Fields FJ, Torres-Tiji Y, Heredia V, Mayfield SP. Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii. PLoS One. 2021 Nov 18;16(11):e0257089. doi: 10.1371/journal.pone.0257089. PMID: 34793485; PMCID: PMC8601568.

From Anthony Berndt, Stephen Mayfield Lab, University of California, San Diego, November 2021

Source: Anthony Berndt, Stephen Mayfield Lab, University of California, San Diego
Plasmid: pRMC3
Vector: pUC19-based backbone
Host strain: DH5alpha
Origin: Gibson assembly using pUC19 backbone from synthesized gBlocks and elements of the pOpt vectors from the Kruse Lab.
Insert: ER-Golgi retained SARS-CoV-2 Spike RBD fused to mClover fluorescent protein.
Selection in E. coli: Ampicillin
Comment: Transformed in to algae strain RMC3. pRMC3 map in GenBank format also available in publication’s supplemental data.

Sequence


Berndt AJ, Smalley TN, Ren B, Simkovsky R, Badary A, Sproles AE, Fields FJ, Torres-Tiji Y, Heredia V, Mayfield SP. Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii. PLoS One. 2021 Nov 18;16(11):e0257089. doi: 10.1371/journal.pone.0257089. PMID: 34793485; PMCID: PMC8601568.