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

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 111508 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

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
  • 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

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.

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