Abstract: Methods and compositions for bacterial production of pure single-stranded DNA (ssDNA) composed of custom sequence and size have been developed. The methods enable scalability and bio-orthogonality in applications of scaffolded DNA origami, offering one-step purification of large quantities of pure ssDNA amendable for immediate folding of DNA nanoparticles. The methods produce pure ssDNA directly from bacteria. In some embodiments the E. coli helper strain M13cp combined with a phagemid carrying only an f1-origin allows for, without the need for additional purification from contaminating dsDNA. This system is useful for generalized circular ssDNA synthesis, and here is applied to the assembly of DNA nanoparticles folded both in vitro and direct from phage.

Abstract: This invention relates to processes that transcribe DNA molecules containing non-standard nucleotides using variants of T7 RNA polymerase to give RNA transcripts that contain their complementary non-standard nucleotides. Non-standard nucleotides pair during transcription using patterns of hydrogen bonding that are different from patterns that join the thymine-adenine and guanine-cytosine nucleobase pairs.

Abstract: Provided herein are constructs for genome editing or genetic engineering its fungi or protists, methods of using the constructs and media for use in selecting cells. The construct include a polynucleotide encoding a thymidine kinase operably connected to a promoter, suitably a constitutive promoter; a polynucleotide encoding an endonuclease operably connected to an inducible promoter; and a recognition site for the endonuclease. The constructs may also include selectable markers for use in selecting recombinations.

Abstract: A shuttle vector is provided which can be manipulated in various kinds of host cells, thereby providing a novel tool for the field of genetic engineering. Also provided are a prokaryotic host cell and a kit including said shuttle vector, so as to construct expression vectors which contain the target gene using the shuttle vector, thereby producing proteins in various host cells with one single vector.