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Addgene

pGGDestTol2LC-5sgRNA
(Plasmid #64243)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 64243 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pDestTol2LC
  • Backbone size (bp) 7083
  • Modifications to backbone
    One BsaI site in ampicillin sequence is mutated. Two BsaI sites containing adaptors with appropriate overhang (CAGT and GAAC) were sequentially inserted into the NsiI/XhoI and KpnI sites of pDestTol2LC.
  • Vector type
    CRISPR

Growth in Bacteria

  • Bacterial Resistance(s)
    Chloramphenicol and Ampicillin, 25 & 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    ccdB Survival
  • Copy number
    High Copy

Cloning Information

  • Cloning method Restriction Enzyme

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    The destination vector for these U6x:sgRNA cassettes were made first by eliminating the BsaI site in pDestTol2CG2 (Chi-Bin Chien lab), followed by replacing the cmcl2:EGFP (AatII/KpnI) with a cryaa:mCerulean (SacII/KpnI) from phsp70l-loxP-mCherry-STOP-loxP-H2B-GFP_cryaa-cerulean to generate pDestTol2LC (Didier Y. R. Stainier lab)
  • Article Citing this Plasmid

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.
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:

    pGGDestTol2LC-5sgRNA was a gift from Wenbiao Chen (Addgene plasmid # 64243 ; http://n2t.net/addgene:64243 ; RRID:Addgene_64243)
  • For your References section:

    Multiplex Conditional Mutagenesis Using Transgenic Expression of Cas9 and sgRNAs. Yin L, Maddison LA, Li M, Kara N, LaFave MC, Varshney GK, Burgess SM, Patton JG, Chen W. Genetics. 2015 Apr 8. pii: genetics.115.176917. 10.1534/genetics.115.176917 PubMed 25855067