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

• Purpose: a nuclear-targeted, genetically-encoded sensor for lactate
• Depositing Lab: Luis Felipe Barros
• Depositing Organization: Centro de Estudios Cientificos
  Located in Chile
• 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/ .

DNA (Catalog # 46307-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 # 46307-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.