pGC479
(Plasmid #42601)

• Depositing Lab: Jane Hubbard
• Publication: Korta et al Development. 2012 Mar;139(5):859-70. doi: 10.1242/dev.074047. Epub 2012 Jan 25.

Backbone

• Vector backbone: L4440
• Backbone size w/o insert (bp): 2790
• Total vector size (bp): 4008
• Vector type: RNAi

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin and Tetracycline, 100 & 10 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Grow on LB + Amp + Tet plates. For RNAi feeding, grow LB liquid culture with Amp only.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: daf-15
• gRNA/shRNA sequence: daf-15
• Species: C. elegans (nematode)
• Entrez Gene: daf-15 (a.k.a. CELE_C10C5.6)
• Promoter: T7

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EagI (not destroyed)
• 3′ cloning site: PstI (destroyed during cloning)
• 5′ sequencing primer: M13F

Terms and Licenses

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

Depositor Comments

This plasmid can be kept in DH5alpha cells but does not work for RNAi in that strain. For RNAi, please transform DNA into the HT115 strain.

Note: Addgene's quality control sequencing has found that the deposited clone contains a T to C mismatch with respect to the C. elegans N2 reference daf-15 sequence, which would cause a V797A change in the coded protein. According to the depositing lab, this point mutation does not affect RNAi feeding; however, its possible effects in other contexts are not known.