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

• Purpose: a genetically-encoded sensor for lactate concentration, transport and metabolic flux
• Depositing Lab: Luis Felipe Barros
• Depositing Organization: Centro de Estudios Cientificos
  Located in Chile
• Publication: San Martin et al PLoS One. 2013;8(2):e57712. doi: 10.1371/journal.pone.0057712. Epub 2013 Feb 26.
• Citations: 41 Articles

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

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/ .

DNA (Catalog # 44238-D50)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 50 μg
• Concentration: 1 μg/μL
• Pricing: $495 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

DNA (Catalog # 44238-D100)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 100 μg
• Concentration: 1 μg/μL
• Pricing: $585 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.