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pSAM_Ec
(Plasmid #102939)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 102939 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)
    Ampicillin and Kanamycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    Pir1
  • Growth instructions
    EcS17/pSAM-Ec need ~36-48 hours to reach an OD600 of 1.0.
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    mariner transposon flanked by MmeI modified inverted repeats and the himar1C9 transposase

Resource Information

  • Supplemental Documents
  • A portion of this plasmid was derived from a plasmid made by
    pSAM_Bt: Goodman AL, McNulty NP, Zhao Y, Leip D, Mitra RD, Lozupone CA, Knight R, Gordon JI (2009). Identifying genetic determinants needed to establish a human gut symbiont in its habitat. Cell Host Microbe, 6: 279–289.
  • Terms and Licenses
  • Industry Terms
    • Not Available to Industry
  • Articles Citing this Plasmid

Depositor Comments

The Plac promoter from pGFP-Mut3.1 (Clonetech), along with its associated ribosome binding sequence, was amplified via PCR. Engineered 59 BamHI and 39 NdeI restriction sites were used to sub-clone the resulting fragment into a BamHI/NdeI (New England Biolabs) double digested pSAM_Bt vector upstream of the himar1C9 transposase gene. The kanamycin resistance gene from pKD4 was amplified and ligated using the restriction sites MfeI and XbaI, replacing the erythromycin resistance gene ermG in pSAM_Bt. The resulting transposon mutagenesis vector, pSAM-Ec, was stored and propagated in the pir+ E. coli strain EcS17.

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:

    pSAM_Ec was a gift from Matthew Mulvey (Addgene plasmid # 102939 ; http://n2t.net/addgene:102939 ; RRID:Addgene_102939)
  • For your References section:

    Combining quantitative genetic footprinting and trait enrichment analysis to identify fitness determinants of a bacterial pathogen. Wiles TJ, Norton JP, Russell CW, Dalley BK, Fischer KF, Mulvey MA. PLoS Genet. 2013;9(8):e1003716. doi: 10.1371/journal.pgen.1003716. Epub 2013 Aug 22. PGENETICS-D-12-03121 [pii] PubMed 23990803