pRB33
(Plasmid #72867)

• Purpose: template for N-terminal 3x FLAG tag, constitutive Puromycin resistance
• Depositing Lab: Klaus Foerstemann
• Publication: Kunzelmann et al G3 (Bethesda). 2016 Jun 1;6(6):1777-85. doi: 10.1534/g3.116.028241.

Backbone

• Vector backbone: pJet1.2
• Backbone size w/o insert (bp): 2974
• Total vector size (bp): 4503
• Modifications to backbone: contains a copia-Puromycin cassette, followed by the Drosophila mtnDE promoter, flanked by FRT-sites
• Vector type: Insect Expression
• Selectable markers: Puromycin

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: 3xFlag tag
• Species: Synthetic
• Insert Size (bp): 63

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: SpeI (not destroyed)
• 5′ sequencing primer: pJet fwd
• 3′ sequencing primer: pJet rev

Resource Information

• Supplemental Documents:
  • Genomic Tagging in Drosophila cells (C-terminal constructs)
  • Genomic Tagging in Drosophila cells (N-terminal constructs)

Terms and Licenses

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

Depositor Comments

We observe rather poor growth of bacteria with the pJet1.2 backbone, resulting in low yields of the plasmid preps.