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pSLGCV
(Plasmid #49862)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 49862 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pPD95.79
  • Backbone manufacturer
    Andrew Fire, Carnegie Institution for Science
  • Modifications to backbone
    added sequentially the sur-5 promoter [36], PCR-amplified from plasmid pTG96 and then the luciferase gene PCR-amplified from vector pSP-luc+ (Promega). A 3.7 kb fragment upstream of the sur-5 gene containing the promoter, was amplified using primers 5'- ATAAGCTTGCATGCCTGCATTGC-3' and 5'-AGACACCCCGGGCTTTCTGAAAAC-3' (this introduces a Sma I site and removes the start codon for the sur-5 gene). The sur-5 promoter was inserted in the pPD95.79 backbone using appropriate restriction sites (Sph I and Sma I). The luc+ gene was amplified using primers 5'-TCCCGGGAAGCTTTCCATGGAAGAC-3' and 5'-CTAGGGTACCACGGCGATCTTTCC-3'. Sma I and Kpn I were used to insert luc+ downstream of the sur-5 promoter and upstream of and in frame with gfp. The peroxisome tagging sequence is absent from luc+ and the NLS for sur-5 was excluded from pSLGCV, so that LUC+::GFP was expressed in the cytoplasm
  • Vector type
    Worm Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    firefly luciferase luc+ fused to GFP (S65C)
  • Species
    Photinus pyralis
  • Promoter sur-5
  • Tag / Fusion Protein
    • GFP (S65C) (C terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site SmaI (unknown if destroyed)
  • 3′ cloning site KpnI (unknown if destroyed)
  • 5′ sequencing primer 5'-TCCCGGGAAGCTTTCCATGGAAGAC-3'
  • 3′ sequencing primer 5'-CTAGGGTACCACGGCGATCTTTCC-3'
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    vector backbone pPD95.79 from Firelab, Carnegie Institution for Science. pTG96 plasmid (from which promoter was PCR- amplified) obtained from John Yochem and Min Han; University of Colorado, Boulder. vector pSP-luc+ (bought from Promega. Luc+ gene was PCR-amplified from here.
  • Articles Citing this Plasmid

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Although no formal MTAs signed with Firelab or University of Colorado Boulder, we accepted that the material was provided for academic research only.

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:

    pSLGCV was a gift from Anne Glover & Jonathan Pettitt (Addgene plasmid # 49862 ; http://n2t.net/addgene:49862 ; RRID:Addgene_49862)
  • For your References section:

    Bridging the phenotypic gap: real-time assessment of mitochondrial function and metabolism of the nematode Caenorhabditis elegans. Lagido C, Pettitt J, Flett A, Glover LA. BMC Physiol. 2008 Apr 2;8:7. doi: 10.1186/1472-6793-8-7. 10.1186/1472-6793-8-7 PubMed 18384668