pT22Ubm4Eng2bKRPC(#493)
(Plasmid #184091)

• Purpose: cyclofen-inducible fish Eng2b (mut) activation in zebrafish permanent transgenic
• Depositing Lab: Sophie Vriz
• Publication: Amblard et al Commun Biol. 2020 Sep 29;3(1):536. doi: 10.1038/s42003-020-01268-7.

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): 8031
• 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: 4myc-ZfEng2b(WW>KK)-ERT2-P2A-mCherry
• Species: H. sapiens (myc), D. rerio (ZfEng2b), H. sapiens (ERT2), PTV-1 (P2A), Discosoma sp. (mCherry)
• Insert Size (bp): 2763
• Mutation: ZfEng2b: W141K, W144K; ERT2: G400V, M543A, L544A, deleted 1-281;
• Promoter: Zf-Ubi
• Tag / Fusion Protein:
  • 4xMyc (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: AscI (not destroyed)
• 5′ sequencing primer: ubidir
• 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

4myc; ZfEng2b: NP_571115; ERT2: NP_000116; P2A: AIE57569; mCherry: AY678264 (polycistronic mRNA: mCherry cytoplasmic marker of expression) + gCRY:CFP as a marker of transgenesis;