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pC-G418-YR
(Plasmid #61767)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 61767 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pCAMBIA-0380
  • Backbone manufacturer
    CAMBIA
  • Backbone size w/o insert (bp) 6812
  • Total vector size (bp) 10768
  • Modifications to backbone
    The hygromycin gene in ternary vector pC-HYG-YR (Addgene plasmid# 61765) was replaced with nptII gene (Geneticin/G418 resistance) to construct pC-G418-YR.
  • Vector type
    Ternary vector for Agrobacterium mediated transformation of fungi
  • Selectable markers
    Neomycin (select with G418), URA3

Growth in Bacteria

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

Gene/Insert 1

  • Gene/Insert name
    nptII gene
  • Alt name
    Geneticin or G418 resistance gene
  • Species
    E. coli
  • Insert Size (bp)
    795
  • GenBank ID
  • Promoter trpC promoter

Cloning Information for Gene/Insert 1

  • Cloning method Restriction Enzyme
  • 5′ cloning site EcoRI (not destroyed)
  • 3′ cloning site HindIII (not destroyed)
  • 5′ sequencing primer LB-F1 (5'-gtggtgtaaacaaattgacgc-3')
  • 3′ sequencing primer RB-F1 (5'-ggataaaccttttcacgccc-3')
  • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    2 micron origin or replication and URA3 gene for S. cerevisiae
  • Species
    S. cerevisiae (budding yeast)
  • Insert Size (bp)
    2487
  • GenBank ID
    KC562906.1

Cloning Information for Gene/Insert 2

  • Cloning method Restriction Enzyme
  • 5′ cloning site SacII (not destroyed)
  • 3′ cloning site SacII (not destroyed)
  • 5′ sequencing primer ccgcggTTAGTTTTGCTGGCCGCATC
  • 3′ sequencing primer ccgcggCGCTTCCACAAACATTGCTC
  • (Common Sequencing Primers)

Resource Information

  • Supplemental Documents
  • A portion of this plasmid was derived from a plasmid made by
    nptII gene (Geneticin/G418 resistance) was cloned from pCGEN

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Jason Rudd, (Rothamsted research UK) provided Agrobacterium tumfaciens vector pCGEN (pCAMBIA-0380 backbone with nptII gene under Neurospora crassa gpd promoter).

NptII (neomycin phosphotransferase II) gene from pCGEN was recombined into pC-HYG-YR (Addgene plasmid# 61765) through yeast recombineering to replace hygromycin gene and create pC-G418-YR.

Restriction sites EcoRI and HindIII can be used to linearise the vector inside left (LB) and right (RB) borders of the TDNA. Note that the trpC promoter and nptII sequences are present within the EcoRI and HindII cloning sites.

See Supplementary Fig. S1. of associated publication for schematic of single step assembly of gene deletion ternary vectors using yeast recombinational cloning.

Please note that the Addgene verified sequence identified a transposase in the backbone (7145-8353 bp).

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:

    pC-G418-YR was a gift from Ken Haynes (Addgene plasmid # 61767 ; http://n2t.net/addgene:61767 ; RRID:Addgene_61767)
  • For your References section:

    Exploitation of sulfonylurea resistance marker and non-homologous end joining mutants for functional analysis in Zymoseptoria tritici. Sidhu YS, Cairns TC, Chaudhari YK, Usher J, Talbot NJ, Studholme DJ, Csukai M, Haynes K. Fungal Genet Biol. 2015 Jun;79:102-9. doi: 10.1016/j.fgb.2015.04.015. 10.1016/j.fgb.2015.04.015 PubMed 26092796