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pCMVd1-NLS-GAL4-iLight-VP16-T2A-mTagBFP2
(Plasmid #170273)

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Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 170273 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 170273-D50 50 μg of DNA in Tris buffer $495
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DNA 170273-D100 100 μg of DNA in Tris buffer $585
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Backbone

  • Vector backbone
    pQP-Select
  • Backbone manufacturer
    Originally derived from pEGFP-N1 backbone after several modifications in our lab.
  • Backbone size w/o insert (bp) 3427
  • Vector type
    Mammalian Expression
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    NLS-Gal4(1-65)-iLight(human codon optimized)-VP16-T2A-mTagBFP2
  • Species
    Bradyrhizobium sp. (strain ORS 278)
  • Insert Size (bp)
    3154
  • Mutation
    See Comments
  • Promoter CMVd1

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NheI (not destroyed)
  • 3′ cloning site BamHI (not destroyed)
  • 5′ sequencing primer CMV Forward
  • 3′ sequencing primer EBV Reverse
  • (Common Sequencing Primers)

Resource Information

Terms and Licenses

Trademarks:

  • Zeocin® is an InvivoGen trademark.

Depositor Comments

To design iLight, a full-length bacterial phytochrome IsPadC from Idiomarina sp. A28L was used as an original template. First, diguanylyl cyclase effector in IsPadC was removed, then following mutations were introduced: I68F, H80Q, A86T, R90S S242C, E274K, R295H I360V, and L464V. After that, iLight was human codon optimized for mammalian expression.

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:

    pCMVd1-NLS-GAL4-iLight-VP16-T2A-mTagBFP2 was a gift from Vladislav Verkhusha (Addgene plasmid # 170273 ; http://n2t.net/addgene:170273 ; RRID:Addgene_170273)

  • For your References section:

    Single-component near-infrared optogenetic systems for gene transcription regulation. Kaberniuk AA, Baloban M, Monakhov MV, Shcherbakova DM, Verkhusha VV. Nat Commun. 2021 Jun 23;12(1):3859. doi: 10.1038/s41467-021-24212-7. 10.1038/s41467-021-24212-7 PubMed 34162879
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