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

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 175258 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

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.

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