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(Plasmid #112497)


Item Catalog # Description Quantity Price (USD)
Plasmid 112497 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.


  • Vector backbone
  • Backbone manufacturer
    Alexeyev, M.F. (PMID 10337469)
  • Backbone size w/o insert (bp) 1860
  • Total vector size (bp) 4582
  • Modifications to backbone
    Construction of mutagenesis vector pSAM (Sequencing-Adapted Mariner). The E. coli ß-lactamase gene was amplified by PCR from pUC19 (New England Biolabs) using primers MluI Ap 5’ and MluI Ap 3’ (see PMID 19748469, Table S19 for a list of all primers used in this study), and subcloned as an MluI fragment into pKNOCK_Cm (Alexeyev, 1999). Platinum Pfx DNA polymerase (Invitrogen) was used for PCR. The mariner transposon was constructed by PCR amplification of the ermG fragment from pGERM (Salyers et al., 2000) using primers ErmGm 5’ and ErmGm 3’, and introduced into pKNOCK_Ap as a KpnI/NotI fragment to create pMar1m. An alternate construct pMar1, lacking MmeI sites in the transposon inverted repeats, was constructed by PCR amplification of the ermG fragment from pGERM with primers ErmG 5’ and ErmG 3’. The gene encoding Himar1C9 transposase (Lampe et al., 1999) was cloned from pBADC9 (kindly provided by J. Mougous, University of Washington), using primers NdeI Himar1C9 5’ and NotI Himar1C9 3’, as an NdeI/Not fragment to create pSAM. The 300bp region upstream of the Bacteroides thetaiotaomicron VPI-5482 rpoD gene (BT1311) was PCR-amplified using primers Bt1311 prom 5’ and Bt1311 prom 3’ and mobilized as an NdeI/BamHI fragment to create pMar3m_Bt1311. To introduce the Illumina P7 sequencing adapters into the transposon, the ermG fragment of pMar3m_Bt1311 was PCR-amplified with P7 5’ and P7 3’ and re-cloned into the vector backbone. An E. coli rrnB T1/T2 transcriptional terminator cassette was amplified from pFW11 (Whipple, 1998) (kindly provided by Simon Dove, Harvard Medical School) using primers XbaI Term 5’ and PstI Term 3’ and cloned as an XbaI/PstI fragment to create pSAM_Bt. Intermediate and final clones were verified by bi-directional sequencing.

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
  • Growth Strain(s)
    S-17 λ pir
  • Copy number

Gene/Insert 1

  • Gene/Insert name
    INSeq-adapted mariner transposon
  • Insert Size (bp)

Gene/Insert 2

  • Gene/Insert name
  • Insert Size (bp)

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
  • Zeocin® is an InvivoGen trademark.
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_Bt was a gift from Andrew Goodman & Jeffrey Gordon (Addgene plasmid # 112497 ; ; RRID:Addgene_112497)
  • For your References section:

    Identifying genetic determinants needed to establish a human gut symbiont in its habitat. Goodman AL, McNulty NP, Zhao Y, Leip D, Mitra RD, Lozupone CA, Knight R, Gordon JI. Cell Host Microbe. 2009 Sep 17;6(3):279-89. doi: 10.1016/j.chom.2009.08.003. 10.1016/j.chom.2009.08.003 PubMed 19748469