pBCN39-R4R3
(Plasmid #34914)

• Purpose: (Empty Backbone)
• Depositing Lab: Ben Lehner
• Publication: Semple et al Nat Methods. 2012 Jan 30;9(2):118-9. doi: 10.1038/nmeth.1864.

Backbone

• Vector backbone: pBCN21-R4R3
• Modifications to backbone: Contains the dual antibiotic resistance operon Prpl-28::PuroR::rpl-16_outron::NeoR::let-858_3'UTR instead of the puromycin resistance gene. Spe I, Xcm I, Bgl II, Avr I and Sac I restriction sites introduced into backbone for linearisation before bombardment. The ccdB gene was replaced with a version from pDONR221 (Invitrogen) that is compatible with commercially available ccdB Survival E. coli, no longer requiring DB3.1 cells.
• Vector type: Worm Expression
• Selectable markers: Neomycin (select with G418), Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Ampicillin, 25 & 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): ccdB Survival
• Growth instructions: As a Gateway destination vector the empty vector must be grown in ccdB resistant cells such as DB3.1 or ccdB Survival. Use both Ampicillin and Chloramphenicol selection to avoid loss of the Gateway cassette. Once the Gateway cassette has been replaced with the gene of interest select with Ampicillin in standard E. coli strain such as Top10 or DH5alpha.
• Copy number: Unknown

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). Neomycin resistance gene from pEGFP-C1 (Clontech).

Terms and Licenses

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

Depositor Comments

Dual resistance (PuroR-NeoR) vector for drug selection following biolistic bombardment in worms.
This vector does not contain a visual marker on the backbone and can be used when the gene of interest has visual phenotype.
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).