pBCN21-R4R3
(Plasmid #26301)

• Purpose: (Empty Backbone)
• Depositing Lab: Ben Lehner
• Publication: Semple et al Nat Methods. 2010 Aug 22. ():.

Backbone

• Vector backbone: pCG150
• Backbone size (bp): 5879
• Vector type: Worm Expression
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Ampicillin, 25 & 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DB3.1
• Growth instructions: Use DB3.1 E. coli (must be ccdB resistant). Note that ccdB Survival bacteria (Invitrogen) do not work for this plasmid.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: None

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AttR4 (destroyed during cloning)
• 3′ cloning site: AttR3 (destroyed during cloning)
• 5′ sequencing primer: M13 Reverse
• 3′ sequencing primer: M13 (-21) Forward

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Puromycin resistance gene from pBabePuro (Addgene). rpl-28 promoter and let-858 3'UTR sequences from pPD129.57 vector (Addgene).

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Use for puromycin selection of transgenes in C. elegans and related species if transgene has a visual phenotype and Pmyo-2::mCherry expression is undesirable.
The unc-119 gene in pCG150 was replaced by the Prpl-28::PuroR::let-858_3'UTR cassette for puromycin selection in worms.
This is a Gateway 3-fragment compatible destination vector.
It contains AttR4 and AttR3 sites (not AttR1 and AttR2 sites as identified automatically by the Addgene algorithm).