1
Plasmid 34916: pBCN41-R4R3
  • None

  • pBCN21-R4R3
    (Search Vector Database)

  • Worm Expression

  • 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. Contains a new version of Pmyo-2::EGFP::myo-2_3'UTR transgene in the backbone that expresses well in various Caenorhabditis species.

  • Ampicillin and Chloramphenicol

  • ccdB Survival

  • 37

  • 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.

  • Unknown

  • Neomycin, Puromycin

  • 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). EGFP gene from pPD129.57 (Addgene).

  • View sequences (4)
  • View map

  • GenBank file (text/plain)

  • Ben Lehner

  • UBMTA

Comments: 

Dual resistance (PuroR-NeoR) vector for drug selection following biolistic bombardment in worms.
This vector contains a Pmyo-2::EGFP::myo-2_3'UTR pharyngeal marker on the backbone. Worm strains generated with vector can be crossed with strains generated with other visual markers such as in pBCN40-R4R3 which has a red fluorescent pharyngeal marker.
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).

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: Generating transgenic nematodes by bombardment and antibiotic selection. Semple et al (Nat Methods. 2012 Jan 30;9(2):118-9. doi: 10.1038/nmeth.1864. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 34916" in your Materials and Methods section.

Price: US $65

Available to academic and non-profits only