Bxb1-GA-Dox-AIO-TetOn_Ngn2-ISL1-LHX3_Down-Tandem
(Plasmid #250431)

• Purpose: Bxb1-GA donor plasmid with downstream tandem syntax for all-in-one doxycycline-inducible expression of Ngn2, ISL1 and LHX3
• Depositing Labs: Richard Davis, Kate Galloway
• Publication: Blanch-Asensio et al bioRxiv [Preprint]. 2024 Oct 17:2024.10.17.616637. doi: 10.1101/2024.10.17.616637.

Backbone

• Vector backbone: pBR322/pUC
• Backbone size w/o insert (bp): 4700
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: Ngn2-P2A-ISL1-T2A-LHX3
• Alt name: hNIL
• Species: H. sapiens (human)
• Entrez Gene: NEUROG2 (a.k.a. Atoh4, Math4A, NGN2, bHLHa8, ngn-2)
• Entrez Gene: ISL1 (a.k.a. ISLET1, Isl-1)
• Entrez Gene: LHX3 (a.k.a. CPHD3, LIM3, M2-LHX3)
• Promoter: TRE3G

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: n/a

Gene/Insert 2

• Gene/Insert name: TetOn3G rtTA
• Promoter: CAG
• Tag / Fusion Protein:
  • T2A-mTagBFP2-3xNLS (C terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: n/a

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
  • Tet Systems Limited Use Label License
• Industry Terms:
  • Not Available to Industry