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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