Supporters

Our team accomplished great things in the project Chlamylicious over the course of this year’s iGEM competition: We adapted a toolkit for the green alga C. reinhardtii with parts for successful and easy cloning of any transcription unit, which we implemented for the expression of the PET-degrading enzyme PETase by the alga and designed a bioreactor for its cultivation under constant monitoring of all parameters. These were our greatest supporters:

• our host laboratories Experimental Biophysics and Molecular Genetics from the Humboldt University of Berlin, who shared their infrastructure with us
• our supervisors Prof. Dr. Schmitz-Linneweber and Prof. Dr. Hegemann
• the principal investigator Simon Kelterborn, who guided us with his expertise in transformations of Chlamydomonas and CRISPR-aided genetic engineering
• the biological technical assistants Irina, Heide (Biophysics lab), Conny, Thea and Irina (all Genetics lab), who enabled us to launch the wet-lab
• computer scientist Vincent Will, who implemented the design for our website
• Dr. Ralf Steuer, who helped us developing the bioreactor
• Prof. Dr. Ringrose, who coached us on how to create a good scientific poster
• electronic engineer Markus Schulz, who helped designing and wiring the electronics for our bioreactor
• technical engineering student Alejandro Gómez Jiménez from the Technical University of Berlin, who made the CAD-models for all parts of our bioreactor
• the Interdisciplinary Workshop „Matters of Activity“ of Humboldt University, who laser-cut our hardware
• the Mechanical Workshop of the Physics Department of our university
• Dr. Nina Siragusa from Merck KGaA, who presented us to her broad scientific network and supported our outreach
• Arne Kuczmera from the Berliner Wasserbetriebe, who supported our research at the waste water treatment plant

Re-used parts from different sources

Part name	Source
AR	Plasmid p114, Hegemann lab (Biophysics Department, Humboldt Universität zu Berlin)
ARS	Ordered oligos from IDT Sequence: A8ISJ6[1-26] (Uniprot)
AmpOri	Amplified from MoClo plasmid pICH47732 (Addgene)
Ble/Scp:	Ordered oligos from IDT
GLE	Sequence: Q93WN9[1 - 28] (Uniprot)
3xHA-Tag	Ordered oligos from IDT
His-Tag	Ordered oligos from IDT
Hyg	Amplified from p360, Hegemann lab (Biophysics Department, Humboldt Universität zu Berlin)
MHETase (wildtype)	2019 iGEM team University of Kaiserslautern, (C. reinhardtii)
MHETase (wildtype II)	Weber lab (Molecular Structural Biology, University of Greifswald), (E. coli)
MHETase (Mutant W397A)	Weber lab (Molecular Structural Biology, University of Greifswald), (E. coli)
Paro	Amplified from p114, Hegemann lab (Biophysics Department, Humboldt Universität zu Berlin)
PETase	Ordered oligos from IDT, Codon usage is for C. reinhardtii
PETase (E. coli)	Adgene's plasmid: plasmid #112203
PtxD	Sequence: AF061070.1 (ncbi) Ordered from GeneCust
PsaD	Plasmid 51, Hegemann lab (Biophysics Department, Humboldt Universität zu Berlin)
PsadIntron	Self designed, intron designed after those of the AR-Promoter
RbcS2	Amplified from p141 pCrP2-SaCas9 (RBCS2, G), p016
RFP	p42, Hegemann lab (Biophysics Department, Humboldt Universität zu Berlin)
Spec	Amplified from the MoClo Plasmid pAGM9121 (Addgene)
YFP	Amplified from YFP Sequenz of p135 pCr3-Ble2-YFP-Sp-APH plasmid pAGM9121 Overhang PCR: Amplification

Timeline and responsibilities

As the first team from the Humboldt University ever to form and participate in the iGEM competition we had to organize the following to establish C. reinhardtii as a chassis for synthetic biology in the iGEM community:

• Find a professor and department able to supervise the iGEM project. As there is no course of synthetic biology taught at our university we referred to the Molecular Genetics and Biophysics lab groups.
• Found a team of students, who were able to conduct the project research during their studies without earning credits for it. Our university so far does not offer any courses for the participation of students as an iGEM team. By June 2018 the main core of our iGEM team was brought together.
• Brainstorm for interesting and viable SynBio projects, which took place in July 2018. Most of us had little or no prior knowledge in synthetic biology.
• Research more on the possibilities of C. reinhardtii as a SynBio tool and familiarize ourselves with project management conducts from July-October 2018. 
• Start fundraising initiatives in July 2018. 
• Get part of the team started with working in the wet lab in October 2018.
• Organize Human Practice events and possibilities of outreach to the broader public from November 2018 - October 2019.
• Get a second lab team to finish the wet lab work from April-October 2019.
• Conceptualize a bioreactor for optimal cultivation and build it from June-October 2019.