Patent ID: 8628954
Filing Date: 2014-01-14
Classification: C12N

Abstract:
1. An expression cassette containing a phage promoter and a sequence encoding a target polypeptide, wherein the cassette is presented with the formula P-X-(S) b T-M, wherein P means the sequence of the phage promoter, X means a target sequence, S means translation stop codon, b=1 (present) or b=0 (absent) in the cassette sequence, T means transcriptional termination sequence for promoters other than phage promoters and M means a sequence encoding a selection marker, required for the survival of a host expressing the target polypeptide, and wherein the sequence M, encoding the selection marker, located in the cassette, is a sequence encoding a protein being a selection marker selected from a group comprising markers which introduce antibiotic resistance, and wherein the sequence encoding the selection marker is a sequence that introduces kanamycin resistance, wherein the cassette includes only a single promoter, which single promoter consists of the phage promoter, wherein the schema of the cassette is: wherein the sequence encoding the selection marker encodes aminoglycolyside-3′ phosphotransferase (APH), wherein the sequence encoding the selection marker has a modified, non-coding part at the 5′ end including a Shine-Dalgarno sequence, wherein said Shine-Dalgarno sequence has five altered nucleotides, wherein starting from the 5′ end the altered nucleotides are: thymine to guanine; adenine to guanine; adenine to cytosine; thymine to cytosine; and adenine to guanine.