pT2iU6Apowriter2N(#355)
(Plasmid #184106)

• Purpose: cyclofen-inducible apoptosis activation and imaging (tEosFP) by zebrafish transient transgenesis or mRNA synthesis
• Depositing Lab: Sophie Vriz
• Publication: Feng et al Rep Prog Phys. 2013 Jul;76(7):072601. doi: 10.1088/0034-4885/76/7/072601. Epub 2013 Jun 14.

Backbone

• Vector backbone: pT2iU6 (pT2AL200R150G derivative)
• Backbone manufacturer: modified by M Volovitch from pT2AL200R150 of A Urasaki; Urasaki et al 2006, Genetics 174:639)
• Backbone size w/o insert (bp): 6242
• Vector type: Zebrafish transient transgenesis + in vitro transcription

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: myr-DIAP1-5myc-tEosFP-nls'-P2A-Casp9-ERT2
• Species: Rous Sarcoma V (myr), fruitfly (DIAP1), human (myc, ERT2), rodent Hepacivirus (nls'), PTV-1 (P2A), mouse (Caspase9)
• Insert Size (bp): 3875
• Mutation: DIAP1: deleted 1-2, N19G, N20V, deleted 146-438; ERT2: G400V, M543A, L544A, deleted 1-281;
• Promoter: Zf-Ubi+SP6
• Tag / Fusion Protein:
  • internal tag: 5myc (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: SnaBI (not destroyed)
• 5′ sequencing primer: SP6
• 3′ sequencing primer: SVLrev34

Resource Information

• A portion of this plasmid was derived from a plasmid made by: synthetic ERT2

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • National Institute of Genetics Limited Use Label License for Tol2 System
• Industry Terms:
  • Not Available to Industry

Depositor Comments

myr; DIAP1: NP_001261916; 5myc; tEosFP: AAV54099; nls; P2A: AIE57569; Casp9: NP_056548; ERT2: NP_000116; polycistronic mRNA expressing both inducible apoptosis trigger (Casp9-ERT2) and apoptosis sensor (myr-DIAP1-tEosFP-nls);