Scalable biotechnological production of DNA single strand molecules of defined sequence and length

A method for the recombinant production of DNA single strand molecules, comprising the steps of:

• (1) providing a pseudogene nucleic acid,
• wherein said pseudogene nucleic acid is a nucleic acid that comprises (i) at least one target DNA oligo- or polynucleotide sequence and (ii) at least one cleavage set of two self-cleaving DNA sequences flanking each target DNA oligo- or polynucleotide sequence, wherein said pseudogene nucleic acid comprises one cleavage set per the one target DNA oligo- or polynucleotide sequence, and two self-cleaving DNA sequences are linked in inverted order to each other to form the one cleavage set;
• (2) integrating the pseudogene nucleic acid into a vector;
• (3) transforming bacterial cells in a bacterial culture with said vector
• (4) producing a precursor ssDNA from said vector under bacterial culture conditions, wherein said precursor ssDNA comprises the pseudogene nucleic acid;
• (5) isolating the precursor ssDNA from the bacterial culture;
• (6) digesting the precursor ssDNA under reaction conditions where the self-cleaving DNA sequences become active and cut off the at least one cleavage set;
• (7) separating the at least one target single stranded DNA oligo- or polynucleotide sequence;
• and
• (8) obtaining the at least one target single stranded DNA oligo- or polynucleotide sequence.