URL: http://www.addgene.org/67940
Proper Citation: RRID:Addgene_67940
Bacterial Resistance: Ampicillin
Defining Citation: PMID:25856300
Vector Backbone Description: Backbone Marker:Stratagene; Vector Backbone:pBluescript II -SK; Vector Types:Other, Cryptococcal complementation vector; Bacterial Resistance:Ampicillin
Comments: Contains the safe haven site sequence in the backbone of pBluescript II SK- to enable integration into the C. neoformans genome via homologous recombination, in addition to a linker sequence containing multiple rare-cutting restriction enzyme cut sites to facilitate linearization prior to biolistic transformation. Importantly, as the safe haven and marker fragments were inserted into the pBluescript II SK- backbone in place of the f1 origin of replication, each plasmid still retains the original multicloning site and blue/white screening capabilities. Linearization of the safe haven constructs is made possible by the inclusion of a linker containing cut sites for three rare cutting restriction enzymes—BaeI, AscI and PacI. To use the vectors, the gene of interest is subcloned into the standard pBluescript II SK- multicloning site. If possible BaeI should be used, as it integrates into the safe haven site approximately 60% of the time, while AscI and PacI only 10-16%.
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Source: Addgene
