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pPV454
(Plasmid #42922)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 42922 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pCR4-TOPO
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 3956
  • Total vector size (bp) 9143
  • Vector type
    Worm Expression
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin and Kanamycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    ccdB Survival
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    Ce-age-1 promoter (873 bp)::Ce-age-1 cDNA (3549 bp)::Ce-unc-54 terminator (765 bp)
  • Alt name
    Age-1
  • Species
    C. elegans (nematode)
  • Insert Size (bp)
    5187
  • GenBank ID
    NM_064061.4 NP_496462.2
  • Entrez Gene
    age-1 (a.k.a. CELE_B0334.8)
  • Promoter Ce-age-1 promoter (873 bp)

Cloning Information

  • Cloning method TOPO Cloning
  • 5′ sequencing primer M13_pUC_fwd; T7
  • 3′ sequencing primer M13_pUC_rev; T3
  • (Common Sequencing Primers)

Resource Information

  • Supplemental Documents
  • A portion of this plasmid was derived from a plasmid made by
    Ce-age-1 cDNA and Ce-unc-54 3' region (terminator) were amplified from the plasmid Pdpy-30-age-1 from Yuichi Iino, University of Tokyo, Tokyo, Japan.
  • Terms and Licenses

Depositor Comments

The construct Ce-age-1p::Ce-age-1(3,549bp)::Ce-unc-54t, contained in plasmid pPV454, was assembled by overlap extension PCR. An 873 bp segment of Ce-age-1 5′ region was amplified from genomic DNA and fused to 2,376 bp of the 5′ region of Ce-age-1, amplified from cDNA, using the overlapping primers Ceage1p-cDNA-F and -R (below). This was subsequently fused to the remainder of the Ce-age-1 cDNA with the plasmid Pdpy-30-age-1, a gift from Yuichi Iino (University of Tokyo, Tokyo, Japan), as template and using the overlapping primers Ceage1-Cat-F and -R (below). Finally, the promoter and full-length 3,549 bp Ce-age-1 cDNA were fused to 765 bp of the Ce-unc-54 3′ region (terminator), amplified from Pdpy-30-age-1 template, using the overlapping primers Ceage1-unc54-F and -R (below). The PCR product was T/A cloned into the pCR4-TOPO vector (Life Technologies) and the insert fully sequenced.

Ceage1p-cDNA-F caagctttcattttaagattttaagATGTCTATGGGACGAAGCCCCTC
Ceage1p-cDNA-R GAGGGGCTTCGTCCCATAGACATcttaaaatcttaaaatgaaagcttg
Ceage1-Cat-F CCTCCGTGGAAATGAAGAGCACatcaagatcatcacccgacaag
Ceage1-Cat-R cttgtcgggtgatgatcttgatGTGCTCTTCATTTCCACGGAGG
Ceage1-unc54-F CCACGCAGTCAAACACTACTGAgataagagctccgcatcggccg
Ceage1-unc54-R cggccgatgcggagctcttatcTCAGTAGTGTTTGACTGCGTGG

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:

    pPV454 was a gift from James Lok (Addgene plasmid # 42922)
  • For your References section:

    Strongyloides stercoralis age-1: a potential regulator of infective larval development in a parasitic nematode. Stoltzfus JD, Massey HC Jr, Nolan TJ, Griffith SD, Lok JB. PLoS One. 2012;7(6):e38587. doi: 10.1371/journal.pone.0038587. Epub 2012 Jun 6. 10.1371/journal.pone.0038587 PubMed 22701676