What are the requirements for my custom vector?
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.
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