pBCN43-R4R3
(Plasmid #34918)

• 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: 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. Contains a new version of Pmyo-2::mCherry::myo-2_3'UTR transgene in the backbone that expresses well in various Caenorhabditis species.
• 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): 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). mCherry gene from pCFJ104 (Addgene).

Terms and Licenses

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

Depositor Comments

New version of Puromycin resistance vector for drug selection in worms.
This vector replaces pBCN22-R4R3 (main advantage of the new vector is compatibility with ccdB Survival cells, and the visual marker that expresses well in non-elegans Caenorhabditis species).
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).