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pPB-VPR-dSpCas9[StaPL(TI)]
(Plasmid #111508)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 111508 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 111508-D50 50 μg of DNA in Tris buffer $495
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DNA 111508-D100 100 μg of DNA in Tris buffer $585
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Backbone

  • Vector backbone
    pPB
  • Backbone manufacturer
    System Biosciences
  • Total vector size (bp) 13958
  • Vector type
    Mammalian Expression
  • Selectable markers
    Puromycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    VPR-dSpCas9[StaPL(TI)]
  • Species
    H. sapiens (human); Hepatitis C Virus (HCV) genotype 1a, Entacmaea quadricolor, Streptococcus pyogenes, Herpes simplex virus, Epstein-Barr virus
  • Insert Size (bp)
    7323
  • Mutation
    The HCV NS3 protease carries F43L, Q80K, and S122R, and D168Y mutations. The VPR domain's internal NLS sequence was mutated to a GGSGGSGGS linker. SpCas9 contains D10A and H840A mutations.
  • GenBank ID
    NC_004102
  • Promoter PGK

Cloning Information

  • Cloning method Gibson Cloning
  • 5′ sequencing primer CCACCTTCTACTCCTCCC
  • 3′ sequencing primer ATTAACCCTCACTAAAGGG
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    the Vladislav Verkhusha lab [TagBFP, Ref. 1], the George Church lab [VPR, Ref. 2], and the Stanley Qi lab [dSpCas9, Ref. 3].

Terms and Licenses

Trademarks:

  • Zeocin® is an InvivoGen trademark.

Depositor Comments

[Ref. 1]
SUBACH O.M., GUNDOROV I.S., YOSHIMURA M., SUBACH F.V., ZHANG J., GRUENWALD D., SOUSLOVA E.A., CHUDAKOV D.M., & VERKHUSHA V.V. 2008. Conversion of Red Fluorescent Protein into a Bright Blue Probe. Chem Biol 15 (10):1116-24.

[Ref. 2]
CHAVEZ A., SCHEIMAN J., VORA S., PRUITT B.W., TUTTLE M., P R IYER E., LIN S., KIANI S., GUZMAN C.D., WIEGAND D.J., TER-OVANESYAN D., BRAFF J.L., DAVIDSOHN N., HOUSDEN B.E., PERRIMON N., WEISS R., AACH J., COLLINS J.J. & CHURCH G.M. 2015. Highly efficient Cas9-mediated transcriptional programming. Nat Methods 12: 326-328.

[Ref. 3]
GAO Y., XIONG X., WONG S., CHARLES E.J., LIM W.A. & QI L.S. 2016. Complex transcriptional modulation with orthogonal and inducible dCas9 regulators. Nat Methods 13: 1043-1049.

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:

    pPB-VPR-dSpCas9[StaPL(TI)] was a gift from Michael Lin (Addgene plasmid # 111508 ; http://n2t.net/addgene:111508 ; RRID:Addgene_111508)

  • For your References section:

    StaPLs: versatile genetically encoded modules for engineering drug-inducible proteins. Jacobs CL, Badiee RK, Lin MZ. Nat Methods. 2018 Jul;15(7):523-526. doi: 10.1038/s41592-018-0041-z. Epub 2018 Jul 2. 10.1038/s41592-018-0041-z PubMed 29967496
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