Ensure your vector meets the following requirements:

• Vector Volume ≥ 100 uL (Plasmid material cannot be submitted on filter paper)
• Vector Concentration ≥ 100 ng/uL
• Vector Mass 10 ug
• Must have bacterial origin of replication (not low copy)
• Must be clonal population
• Must be circular
• Must have antibiotic resistance marker
• Must not contain homopolymers/repetitive sequence >9bp within 40bp of the insertion site
• Repeats: Must contain at least 40bp of unique sequence with no direct or inverted repeats >19bp upstream and downstream of the insertion site
• Must not contain ccdB in final gene product
• Must be able to linearize by Restriction Enzyme site within 50bp of insertion site
• Must be BSL-1 compliant

Antibiotic Resistance:

Plasmid must contain one of the below antibiotic resistance markers for E.coli. Standard concentrations are below:

Antibiotic	Standard concentration available in Production (ug/mL)
Kan	50
AMP	100
CHLOR	25
APRA	50
SPEC	100
TET	10
ZEO	50
BLAST	100

Cell Lines (E. coli)       

• Cell Line available in Production        
• Cloning strain similar to DH10Beta   
• Stbl3-like strain (can be grown at 37℃)        

Copy number              

Only constitutive origins of replication are allowed.

Twist Definition	Accepted	Example
High	Yes	ColE1/pMB1/pBR322/pUC
Medium	Yes, reduced miniprep yield	p15A
Low	No	pSC101

Growth Conditions     

• Standard Temperature: 37℃/30℃   
• Standard Time: Overnight/24hrs         
• Plated colonies must reach an automation friendly size after 16hrs/24hrs of growth on LB-Lennox.