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pTub-alpha1-iLifeact-NeonGreen
(Plasmid #175258)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 175258 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 175258-D50 50 μg of DNA in Tris buffer $495
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DNA 175258-D100 100 μg of DNA in Tris buffer $585
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Backbone

  • Vector backbone
    pTub-alpha1-EGFP
  • Backbone size w/o insert (bp) 4482
  • Vector type
    Mammalian Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    NEB Stable
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    iLifeact
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    783
  • Mutation
    None (wt)
  • Promoter pTub-alpha1
  • Tag / Fusion Protein
    • mNeonGreen (C terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site HindIII (not destroyed)
  • 3′ cloning site NotI (not destroyed)
  • 5′ sequencing primer pTub-alpha1 ( 5'-GTAAGAAGCAACACCTCCTC-3')
  • 3′ sequencing primer NeonGFP reverse (5’-GGAGAGAGGCCATGTTATCC-3’)
  • (Common Sequencing Primers)

Resource Information

Terms and Licenses

Trademarks:

  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Improved (i)LifeAct-NeonGreen is prepared by mutating the start codon of the fluorophore ATGxCTG leaving a unique Kosak sequence-enhanced ATG start codon preceding the LifeAct. We have experimentally detected in previous LifeAct versions that polymerase tend to skip LifeAct translation and start translation at the fluorophore instead, increasing fluorophore-derived background.

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

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.

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

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.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pTub-alpha1-iLifeact-NeonGreen was a gift from Frank Bradke & Sebastián Dupraz (Addgene plasmid # 175258 ; http://n2t.net/addgene:175258 ; RRID:Addgene_175258)

  • For your References section:

    In Situ Visualization of Axon Growth and Growth Cone Dynamics in Acute Ex Vivo Embryonic Brain Slice Cultures. Alfadil E, Bradke F, Dupraz S. J Vis Exp. 2021 Oct 14;(176). doi: 10.3791/63068. 10.3791/63068 PubMed 34723948
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