NLS-Laconic/pCMV-myc-nuc
(Plasmid #46307)

• Purpose: a nuclear-targeted, genetically-encoded sensor for lactate
• Depositing Lab: Luis Felipe Barros
• Publication: Barros et al Front Cell Neurosci. 2013;7:27. doi: 10.3389/fncel.2013.00027. Epub 2013 Mar 22.

Backbone

• Vector backbone: pCMV/myc/nuc
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5000
• Total vector size (bp): 7364
• Vector type: Mammalian Expression ; lactate biosensor
• Selectable markers: Neomycin (select with G418)

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: NLS-Laconic
• Alt name: nuclear-targeted Laconic (LACtate Optical Nano Indicator from CECs)
• Species: Synthetic
• Insert Size (bp): 2230
• Promoter: CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Xho I (not destroyed)
• 3′ cloning site: Not I (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: BGH-rev

Terms and Licenses

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

Depositor Comments

Laconic is a genetically-encoded biosensor that detects lactate in the physiological range. The sensor was based on LldR, a bacterial transcription regulator that consists of a lactate-binding/regulatory domain and a DNA-binding domain. This plasmid expresses a nuclear targeted Laconic via a N-terminal nuclear localization signal (NLS). The laconic sensor consists of a N-terminal mTFP, followed by the LldR flanked by linkers, and finally a C-terminal Venus.

See Addgene plasmid# 44238: Laconic/pcDNA3.1(-) for a non-targeted version of Laconic: http://www.addgene.org/44238/ .