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URL: http://www.addgene.org/26095

Proper Citation: RRID:Addgene_26095

Bacterial Resistance: Ampicillin

Vector Backbone Description: Backbone Marker:Structural Genomics Consortium; Vector Backbone:pFHMSP-LIC-N; Vector Types:Insect Expression, Baculovirus expression; Bacterial Resistance:Ampicillin

Comments: The pFHMSP-LIC N vector is a derivative of the pFastBac HT A vector (Invitrogen). Honeybee melittin signal sequence was introduced before His tag. It is a donor vector for generation of recombinant baculovirus by site-specific transposition into a baculovirus shuttle vector (bacmid) in E. coli host strain, DH10Bac. For use in Bac-to-Bac Baculovirus Expression System in insect cells for secreted protein expression. This vector adds a 26 amino acid N-terminal fusion tag containing 6X His followed by a TEV cleavage site. Insertion of DNA sequence into the cloning/expression region is performed using BD-Biosciences Infusion enzyme mediated by directional recombination between complementary nucleotide DNA sequences at the ends of the insert (PCR product) and Nco1/HindIII linearized vector. Insertion of target sequence involves replacement of a SacB gene stuffer sequence, which provides for negative selection of the original plasmid on 5% sucrose. http://www.sgc.utoronto.ca/SGC-WebPages/toronto-vectors.php

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