Plasmid 26302: pBCN22-R4R3

• Gene/insert name: None
• Vector backbone: pCG150
  (Search Vector Database)
• Vector type: Worm Expression
• Backbone size (bp): 7401
• Cloning site 5': AttR4
• Site destroyed during cloning: Yes
• Cloning site 3': AttR3
• Site destroyed during cloning: Yes
• 5' sequencing primer: M13 Reverse
• 3' sequencing primer: M13 (-21) Forward
• Bacterial resistance(s) Ampicillin and Chloramphenicol
• Growth strain(s) DB3.1
• Growth temperature (℃): 37
• Growth instructions: Use DB3.1 E. coli (must be ccdB resistant). Note that ccdB Survival bacteria (Invitrogen) do not work for this plasmid.
• High or low copy: Low Copy
• Selectable markers: Puromycin
• Person or lab that originally
  cloned the gene/insert: Puromycin resistance gene from pBabePuro (Addgene). rpl-28 promoter and let-858 3'UTR sequences from pPD129.57 vector (Addgene). mCherry gene from pCG144 (Seydoux lab, Addgene). Pharyngeal enhancer (C183) with minimal myo-2 promoter amplified from genomic DNA of AZ218 worms (Caenorhabditis Genetics Center).
• Sequence: View sequences (3)
• Map: View map
• Principal Investigator: Ben Lehner
• Terms and Licenses: MTA

Comments: 

Use for puromycin selection of transgenes in C. elegans.
This vector is identical to pBCN21-R4R3, but a 8xC183_minPmyo-2::mCherry::unc-54_3'UTR cassette was inserted in the backbone as an additional visual 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).

Article: Rapid selection of transgenic C. elegans using antibiotic resistance. Semple et al (Nat Methods. 2010 Aug 22. ():. PubMed)