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URL: http://www.addgene.org/128361
Proper Citation: RRID:Addgene_128361
Insert Name: Acetamidase (amdS)
Organism: Synthetic
Bacterial Resistance: Ampicillin
Defining Citation: PMID:31220607
Vector Backbone Description: Backbone Marker:Stratagene; Vector Backbone:pBlueScript II SK(-); Vector Types:Unspecified; Bacterial Resistance:Ampicillin
Comments: Contains the Safe Haven Site 1 sequence (CNAG_00777 - CNAG_00778) in the backbone of pBluescript II SK- flanked by blaster repeats 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.
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Source: Addgene
