⚠️ Biosafety Guidelines

Essential safety protocols for synthetic biology research

Biosafety Levels (BSL)

All biological materials must be handled according to appropriate biosafety levels:

🟢 BSL-1

Minimal Risk

Non-pathogenic E. coli strains
S. cerevisiae (baker's yeast)
B. subtilis
Standard lab practices
Open bench work allowed

🟡 BSL-2

Moderate Risk

Human cell lines
Viral vectors (AAV, lentivirus)
S. aureus
Biological safety cabinet required
Limited access area

🔴 BSL-3

High Risk

M. tuberculosis
SARS-CoV-2
Yellow fever virus
Specialized facilities
Restricted access

⚫ BSL-4

Maximum Risk

Ebola virus
Marburg virus
Lassa fever
Full containment suits
Specialized facilities only

🚫 Prohibited Sequences

The following sequences are NOT permitted on the platform:

Select Agents: Any sequence from the CDC/USDA Select Agent list
Toxins: Botulinum toxin, ricin, abrin, tetrodotoxin
Virulence Factors: Unmodified pathogenicity islands
Drug Resistance: Novel antibiotic resistance cassettes without justification
Human Pathogens: Complete genomes of human pathogens
Oncogenes: Activated oncogenes without safety controls
Prions: Prion protein sequences (PrP)

Common Restriction Sites Safety

When designing plasmids, consider these common restriction enzymes and their safety implications:

Enzyme	Recognition Site	Safety Note	Common Use
EcoRI	GAATTC	Safe, standard	General cloning
BamHI	GGATCC	Safe, standard	Expression vectors
PstI	CTGCAG	Safe, standard	Blue-white screening
XbaI	TCTAGA	Safe, BioBrick compatible	Synthetic biology
SpeI	ACTAGT	Safe, BioBrick compatible	Modular assembly
NotI	GCGGCCGC	Safe, rare cutter	Large DNA fragments

AAV Safety Considerations

For Adeno-Associated Virus (AAV) vectors:

Pre-Submission Checklist

Replication-incompetent vector confirmed

No wild-type AAV contamination

Transgene safety verified (no oncogenes)

Serotype clearly documented

Tropism and biodistribution data included

BSL-2 containment procedures followed

Antibody Safety Guidelines

When registering antibodies:

Source Documentation: Clearly indicate if derived from human, mouse, rabbit, or other species
Hybridoma Lines: Ensure cell lines are mycoplasma-free
Therapeutic Antibodies: Include any known immunogenicity data
Bispecific Antibodies: Document both target specificities
Conjugated Antibodies: Declare any toxic payloads (e.g., drug conjugates)
Storage Conditions: Specify required storage (e.g., -20°C, -80°C)

Gene Drive & CRISPR Safety

⚠️ Special Restrictions

Gene Drives: NOT permitted without explicit approval
Human Germline Editing: Strictly prohibited
CRISPR in Pathogens: Requires safety justification
Cas Enzymes: Must include PAM sequence documentation
Guide RNAs: Off-target analysis required for human targets

Antibiotic Resistance Markers

Acceptable resistance markers for selection:

✅ Commonly Accepted

Ampicillin (bla)
Kanamycin (kan)
Chloramphenicol (cat)
Tetracycline (tet)
Spectinomycin (spec)
Gentamicin (gent)

⚠️ Use with Caution

Vancomycin resistance
Methicillin resistance
Carbapenem resistance
Multiple drug resistance cassettes

Reporting Safety Concerns

If you identify potentially hazardous sequences or safety issues:

Immediate Action: Do not proceed with tokenization
Contact: Email [email protected] with details
Include: Sequence identifier, specific concerns, BSL level
Wait: For safety team review before proceeding

External Safety Resources