pPV454
(Plasmid #42922)

• Depositing Lab: James Lok
• Publication: Stoltzfus et al PLoS One. 2012;7(6):e38587. doi: 10.1371/journal.pone.0038587. Epub 2012 Jun 6.

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, 100 & 50 μg/mL
• 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

Resource Information

• Supplemental Documents:
  • pPV454.ape
• 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

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

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