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pSEVA3b61
(Plasmid #58310)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 58310 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

Growth in Bacteria

  • Bacterial Resistance(s)
    Chloramphenicol (6 ug/mL)
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH10B
  • Growth instructions
    In E. coli, use chloramphenicol at 6 µg/mL (gene expression of cat not very strong). In B. subtilis, the same concentration is sufficient for selection.
  • Copy number
    High Copy

Resource Information

  • Terms and Licenses
  • Industry Terms
    • Not Available to Industry

Depositor Comments

Despite multiple attempts, we could never clone the cat gene in pSEVA3b61 in the opposite orientation. We concluded that the cat promoter was not sufficient for chloramphenicol resistance in E. coli; rather, the gene was relying on read-through from the upstream repB promoter. We tried several variants extending the cmR promoter region without success. It is important to note that the cat gene works just fine in B. subtilis in either orientation. We found the following paper useful for background reading on this particular cat gene: Olson, D. G., & Lynd, L. R. (2012). Computational design and characterization of a temperature-sensitive plasmid replicon for gram positive thermophiles. Journal of Biological Engineering, 6(1), 5. doi:10.1186/1754-1611-6-5

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pSEVA3b61 was a gift from Tom Ellis (Addgene plasmid # 58310 ; http://n2t.net/addgene:58310 ; RRID:Addgene_58310)
  • For your References section:

    GeneGuard: a Modular Plasmid System Designed for Biosafety. Wright O, Delmans M, Stan GB, Ellis T. ACS Synth Biol. 2014 May 13. 10.1021/sb500234s PubMed 24847673