pT22i5uasm5En25EE4P3Che(#1213)
(Plasmid #184099)

• Purpose: cyclofen-inducible chicken EN2 (mut) activation in zebrafish permanent transgenic
• Depositing Lab: Sophie Vriz
• Publication: Vriz lab cyclofen-inducible ERT2 plasmids (unpublished)

Backbone

• Vector backbone: pT22i (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): 4646
• Vector type: Zebrafish transgenesis (blue eyes)

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: 5myc-chkEn2(5E)-ERT2-5'fP2A-mCherry
• Species: H. sapiens (myc), G. gallus (chkEn2), H. sapiens (ERT2), PTV-1 (5'fP2A), Discosoma sp. (mCherry)
• Insert Size (bp): 2918
• Mutation: chkEn2: S150E, S151E, S153E, S155E, S156E; ERT2: G400V, M543A, L544A, deleted 1-281;
• Promoter: 5xUAS
• Tag / Fusion Protein:
  • 5xMyc (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: ClaI (not destroyed)
• 3′ cloning site: AscI (not destroyed)
• 5′ sequencing primer: insprimA
• 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
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

5myc; chkEn2: NP_001254648; ERT2: NP_000116; 5'fP2A: AIE57569; mCherry: AY678264 (polycistronic mRNA: mCherry cytoplasmic marker of expression) + gCRY:CFP as a marker of transgenesis;