tetO-MITF
(Plasmid #170695)

• Purpose: doxycycline-inducible overexpression of MITF
• Depositing Lab: Paul Burridge
• Publication: Romero-Tejeda et al J Mol Cell Cardiol. 2023 Jul 6:S0022-2828(23)00106-2. doi: 10.1016/j.yjmcc.2023.06.005.

Backbone

• Vector backbone: FU-tetO-Gateway
• Backbone manufacturer: addgene plasmid #43914
• Backbone size w/o insert (bp): 8600
• Total vector size (bp): 10170
• Modifications to backbone: MITF ORF added to plasmid using Gateway cloning
• Vector type: Mammalian Expression, Lentiviral ; Doxycycline Inducible

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: MITF
• Alt name: microphthalmia transcription factor, A variant
• Alt name: MITF variant 1
• Species: H. sapiens (human)
• Insert Size (bp): 1563
• Entrez Gene: MITF (a.k.a. CMM8, COMMAD, MI, MITF-A, WS2, WS2A, bHLHe32)
• Promoter: TRE promoter, Tet-ON

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: LNCX; T7

Resource Information

• A portion of this plasmid was derived from a plasmid made by: pEGFP-N1-MITF-A was a gift from Shawn Ferguson (Addgene plasmid # 38132 ; http://n2t.net/addgene:38132 ; RRID:Addgene_38132)

Terms and Licenses

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

Depositor Comments

This plasmid was generated using Gateway cloning. attB sites were added to MITF via PCR reaction using pEGFP-N1-MITF-A as a template. Resulting fragements were recombined with pDONR221 to generate an entry clone. LR reaction was performed using the destination vector FU-tetO-Gateway (Addgene Plasmid #19778).