Patent ID: 8741633

Claim:
A vector for inserting heterologous genes of interest into a target DNA sequence in the genome of a host cell adjacent to an open reading frame (ORF) comprising 5′ to 3′ with reference to the vector: (a) a first homologous vector DNA sequence capable of homologous recombination with a 5′ flanking region of the target DNA sequence, which first vector DNA sequence includes an open reading frame (ORF) of the target DNA sequence and ends at its 3′ end with a stop codon of the open reading frame (ORF) in the target DNA sequence; (b) a heterologous transcription termination DNA sequence operably linked to the 3′ end of the first vector DNA sequence such that the transcription termination sequence is adjacent to the stop codon for the ORF of part (a); (c) a DNA sequence comprising one or more expression cassettes linked to the 3′ end of the heterologous transcription termination DNA sequence of part (b); and (d) a second homologous vector DNA sequence capable of homologous recombination with a 3′ flanking region of the target DNA sequence, said second homologous vector DNA sequence linked to the 3′ end of the DNA sequence comprising the one or more expression cassettes of part (c); wherein the vector is capable of undergoing homologous recombination with the target DNA sequence to provide an integrated vector construct, and wherein the integrated vector construct creates an intergenic region in the host genome which does not duplicate or delete host genomic sequences and does not disrupt functional expression of the target gene.