pDONOR-MARINA
(Plasmid #223328)

• Purpose: DONOR vector for genomic integration of MARINA voltage sensitive indicator gene from Platisa et al., 2017 (10.1021/acschemneuro.6b00234).
• Depositing Lab: Jonathan Arias
• Publication: Petruškevičiūtė et al Stem Cell Res. 2024 Dec 12:103628. doi: 10.1016/j.scr.2024.103628.

Backbone

• Vector backbone: pDONOR_AAVS1
• Vector type: Mammalian Expression, CRISPR, Synthetic Biology

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: MARINA
• Alt name: Ciona voltage sensitive phosphatease CiVSP derivative from NP_001028998.1
• Species: Synthetic
• Promoter: CAG
• Tags / Fusion Proteins:
  • mCherry (C terminal on insert)
  • super-ecliptic pHluorin (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: pCAG-F

Gene/Insert 2

• Gene/Insert name: KIR2.1 channel Golgi-to-plasma membrane trafficking signal
• Species: H. sapiens (human)
• Promoter: CAG

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: pCAG-F

Resource Information

• Supplemental Documents:
  • 223328_pDONOR-MARINA.gb
• A portion of this plasmid was derived from a plasmid made by: Cas9-based knock-in donor plasmid for the integration of cargo genes into the PPP1R12C gene, also called AAVS1 locus. Gene editing in combination with Addgene plasmid #223321. Carries the genetically encoded reporter MARINA from Platisa et al., 2017 doi:10.1021/acschemneuro.6b00234 which allows monitoring depolarization of human cells.

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Cas9-based knock-in donor plasmid for the integration of cargo genes into the PPP1R12C gene, also called AAVS1 locus. Gene editing in combination with Addgene plasmid #223321. Carries the genetically encoded reporter MARINA from Platisa et al., 2017 doi:10.1021/acschemneuro.6b00234 which allows monitoring depolarization of human cells.