pMinDis.X513-iNicSnFR3a-(CC93)-PM
(Plasmid #125122)

• Purpose: Develop a family of genetically encoded fluorescent biosensors for nicotine, termed iNicSnFRs thus enabling optical subcellular pharmacokinetics for nicotine
• Depositing Lab: Henry Lester
• Publication: Shivange et al J Gen Physiol. 2019 Feb 4. pii: jgp.201812201. doi: 10.1085/jgp.201812201.

Backbone

• Vector backbone: pMinDis.X513
• Backbone manufacturer: Janelia
• Backbone size w/o insert (bp): 5005
• Total vector size (bp): 6832
• Modifications to backbone: NA
• Vector type: Mammalian Expression
• Selectable markers: NA

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Use BL21(DE3) for bacterial expression and grow at 30 degrees C
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: iNicSnFR3a-(CC93)-PM
• Alt name: iNicSnFR3a-(CC93)-PM
• Species: Mammalian
• Insert Size (bp): 1827
• Mutation: NA
• Promoter: T7
• Tags / Fusion Proteins:
  • membrane anchoring sequence-PDGFR (C terminal on insert)
  • Ig-K-Chain (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BglII (unknown if destroyed)
• 3′ cloning site: pstI (unknown if destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: BGH reverse

Resource Information

• Supplemental Documents:
  • pMinDis-X513-iNicSNFR3a(CC93)_PM.gb
• A portion of this plasmid was derived from a plasmid made by: NA

Terms and Licenses

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

Depositor Comments

Please note that a 6bp insertion was identified in the NeoR/KanR cassette during Addgene's quality control process. We recommend testing NeoR selection before performing experiments. Vector was received from Janelia HHMI.