Our laboratory is certified for Biosafety Level 1 (BS-1) work, and we have access to a Biosafety Level 2 lab for BSL2 work. Our work fell within the BSL-1 domain, as indicated by the Center of Disease Control guidelines. All biological waste was stored in autoclave bags and was autoclaved prior to disposal. Sharps and broken glassware were disposed of according to institutional guidelines. Hazardous liquid waste was clearly labeled, and stored in secondary containment for disposal by the institution. Thus, although there is potential for harm to researchers, it is minimized through following procedures approved and used by many laboratories at Yale. It is also minimized by training and common sense. E. coli strains used were common laboratory strains and not pathogenic.

All students working in the laboratory were required to complete the following set of training tutorials, including passing a test at the end.

• http://ehs.yale.edu/training/biosafetypartI
• http://ehs.yale.edu/training/biosafetypartII
• http://learn.caim.yale.edu/chemsafe/login.html
• http://ehs.yale.edu/training/hazardous-chemical-waste-management

All materials were used in accordance with local, national, and Yale Biosafety requirements. Standard lab practices were followed, including secondary containment of chemicals, proper storage of volatiles and flammables, and separation of acids and bases. Nitrile gloves were worn at all times within the lab. A pipet was kept exclusively for ethidium bromide use. Fume hoods were used when handling volatile compounds, concentrated acids and bases, and other reagents. Inhalation and skin contact was avoided. Chemical agents were properly disposed of in designated biohazard waste bins. When UV light was used to visualize gels or GFP, special care was taken to avoid skin or eye exposure. Absolutely no food was allowed in the lab.

Our project was overseen by the Yale Biological Safety Committee and the Office of Environmental Health and Safety (OEHS). Our project has been approved as consistent with Yale's safety regulations. No changes to our project were required since proper protocols were followed. Training was completed as described above.

We do not anticipate any threat to public safety. Organisms worked with are all non-pathogenic. They are likely unable to survive outside of the lab environment, because they will be unable to compete with other organisms in nature. Biomaterials were autoclaved after use. We did not use gloves to touch doors outside of the laboratory to avoid others coming into contact with our chemical and biological agents.

Hands were washed before and after leaving the laboratory.

Our project will ideally be scaled up for medical and industrial use. Possible issues are allergies to the product and if used in immense qualities, DOPA toxicity to the environment. However, proteins and amino acids will be degraded and recycled by the environment, so we do not believe the product will be extremely harmful.

If we continued future work on this project we would assess the risks and hazards associated with it. Once understood, we would attempt to alter the design to minimize these risks. The E. Coli chassis for the product we are designing is not likely to be a great risk on its own.

There are no additional risks posed by our projects compared to other general BSL1 lab concerns. Our bacteria are not pathogenic and are unable to survive outside of the lab environment, because they are unable to effectively compete with other organisms in nature. They do not cause adverse reactions in immunocompetent humans. They do not cause infection. Toxic materials were all disposed of according to Yale waste standards to prevent adverse environmental impact.

There are no safety issues raised by the BioBrick parts submitted to the Registry this year. None of our constructs create toxic gene products for humans or animals.