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URL: http://www.addgene.org/61767
Proper Citation: RRID:Addgene_61767
Insert Name: nptII gene
Organism: E. coli
Bacterial Resistance: Kanamycin
Defining Citation: PMID:26092796
Vector Backbone Description: Backbone Marker:CAMBIA; Backbone Size:6812; Vector Backbone:pCAMBIA-0380; Vector Types:Ternary vector for Agrobacterium mediated transformation of fungi; Bacterial Resistance:Kanamycin
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).
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Source: Addgene
