For all our performed experiments, we have followed the provided iGEM safety rules. The rules that are applicable to our project can be found below:

• We have filled in the Safety Form to provide information on any risks from our projects and steps taken to manage those risks. The Safety Form was checked and signed by our Primary PI.
• Within our project, we do not work with hazardous organisms, parts of activities; all aspects of our project are within the White List Policy. For this reason, we did not have to fill in a Check-In Form.
• Our team has followed all relevant international, regional, national, local or institutional laws, rules, regulations, or policies, including national or institutional biosafety and biosecurity rules applicable to our project.
• As our project can be performed in a ML-I biosafety level, we have only performed experiments with a ML-I lab.
• For a specific part of our experiment, we had to transport our sample to another lab in the Netherlands. To make sure this action is within the "Not Release" category, we had to filter our sample to remove the bacteria completely. To make sure the sample did not contain any bacteria after filtering, we checked for potential growth on an agar plate. No growth was visible. Therefore, we concluded that filling in a Check-In Form was not neccessary, which was confirmed by a iGEM HQ employee.

To strengthen our knowledge about safely working with microorganisms, every member of our team followed the SMT course provided by the TU/e. This course was given by the biological safety officer from the lab where our team will perform their experiments. The course consists of two parts:

• Presentation: in this presentation a safety instruction video was shown. Risk classes for pathogens and risk assessment were explained, as well as methods to prevent aerosols spreading. The different safety equipment (Safety cabinet, Bunsen burner, etc.) were also shown. The last part of the presentation consisted of legislation outside and within the TU/e.
• SMT case studies: every team member was given a document with multiple SMT case studies. These SMT cases consisted of information as well as questions. The goal of these questions was to test the aqcuired knowledge of our members on risk assessment. When these SMT cases were correctly answered, our members received a SMT Certificate.

All experiments executed by iGEM Team ‘Promise’ Eindhoven 2024 follow the safety policies and rules from iGEM, Eindhoven University of Technology and the Dutch government. The organisms, E. Coli strains Top10, dH5α and BL21 and M. Smegmatis strain mc2 155, and the parts, used in the experiments are all on the iGEM 2024 White List. The experiments of the Eindhoven iGEM 2024 team have only been performed in a ML-I lab. Prior to obtaining access to the ML-I lab, all members of the team received training about safely working with GMOs from our university’s biological safety officer. During our project, we kept in close contact with our biological safety officer to discuss the safety of our experiments. On top of that, our lab manager explained to us how to safely work within the lab and gave us a detailed tour around the lab. While working in the lab, the members of our team followed strict safety protocols for personal protection; we did not wear shorts or open shoes in the lab and long hair was always tied. Next to that, we wore a lab coat, glasses and gloves when needed. For disposing of waste, we used multiple different bins. Biological waste was collected on the work bench in red biohazard bag, after which this bag was disposed into the blue container with a yellow lid. Chemical waste was disposed into the black container.