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Plasmid 34917: pBCN42-R4R3
  • None

  • pBCN21-R4R3
    (Search Vector Database)

  • Worm Expression

  • Spe I, Xcm I, Bgl II, Avr I and Sac I restriction sites introduced into backbone for linearisation, if necessary. 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.

  • 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

  • Puromycin

  • Puromycin resistance gene from pBabePuro (Addgene). rpl-28 promoter and let-858 3'UTR sequences from pPD129.57 vector (Addgene).

  • View sequences (3)
  • View map

  • Ben Lehner

  • UBMTA

Comments: 

New version of Puromycin resistance vector for drug selection in worms.
This vector replaces pBCN21-R4R3 (main advantage of the new vector is compatibility with ccdB Survival cells).
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. Full plasmid sequence is available only if provided by the depositing laboratory.

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 34917" in your Materials and Methods section.

Price: US $65

Available to academic and non-profits only