Laconic/pcDNA3.1(-)
(Plasmid #44238)

• Purpose: a genetically-encoded sensor for lactate concentration, transport and metabolic flux
• Depositing Lab: Luis Felipe Barros
• Publication: San Martin et al PLoS One. 2013;8(2):e57712. doi: 10.1371/journal.pone.0057712. Epub 2013 Feb 26.

Backbone

• Vector backbone: pcDNA3.1(-)
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5418
• Total vector size (bp): 7648
• 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: Laconic (LACtate Optical Nano Indicator from CECs)
• Species: Synthetic
• Insert Size (bp): 2230
• Promoter: CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (unknown if destroyed)
• 3′ cloning site: HindIII (unknown if destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: BGH-rev

Resource Information

• Articles Citing this Plasmid:
  • 32 References

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. Laconic consists of a N-terminal mTFP, followed by the LldR flanked by linkers, and finally a C-terminal Venus.

See Addgene plasmid# 46307: NLS-Laconic/pCMV-myc-nuc for a nuclear-targeted version of Laconic: http://www.addgene.org/46307/ .