Methods are provided herein for assembling at least two nucleic acids using a sequence specific nuclease agent (e.g., a gRNA-Cas complex) to create end sequences having complementarity and subsequently assembling the overlapping complementary sequences. The nuclease agent (e.g., a gRNA-Cas complex) can create double strand breaks in dsDNA in order to create overlapping end sequences or can create nicks on each strand to produce complementary overhanging end sequences. Assembly using the method described herein can assemble any nucleic acids having overlapping sequences or can use a joiner oligo to assemble sequences without complementary ends.

Methods are provided herein for assembling at least two nucleic acids using a sequence specific nuclease agent (e.g., a gRNA-Cas complex) to create end sequences having complementarity and subsequently assembling the overlapping complementary sequences. The nuclease agent (e.g., a gRNA-Cas complex) can create double strand breaks in dsDNA in order to create overlapping end sequences or can create nicks on each strand to produce complementary overhanging end sequences. Assembly using the method described herein can assemble any nucleic acids having overlapping sequences or can use a joiner oligo to assemble sequences without complementary ends.

Provided herein are methods of cloning into vectors.

Provided herein are methods of cloning into vectors.

This document provides methods and materials for assembling nucleic acid constructs (e.g., TALENs). For example, methods for assembling TALEs that are rapid, flexible for use in many cloning scaffolds (such as common nuclease and nickase backbones), and achievable with standard molecular biology laboratory tools, thereby making TALEs a more accessible genome system, are provided.

This document provides methods and materials for assembling nucleic acid constructs (e.g., TALENs). For example, methods for assembling TALEs that are rapid, flexible for use in many cloning scaffolds (such as common nuclease and nickase backbones), and achievable with standard molecular biology laboratory tools, thereby making TALEs a more accessible genome system, are provided.

Described herein are methods and related compositions for inducing differentiation of human pluripotent stem cells (hPSCs) into hemogenic endothelium with pan-myeloid potential or restricted potential, by forced expression in the hPSCs of a combination of transcription factors as described herein.

Described herein are methods and related compositions for inducing differentiation of human pluripotent stem cells (hPSCs) into hemogenic endothelium with pan-myeloid potential or restricted potential, by forced expression in the hPSCs of a combination of transcription factors as described herein.

The invention relates to a recombinant Escherichia coli expressing a fusion protein of formamidase and phosphite dehydrogenase, a construction method and use thereof. The invention includes adopting engineered E. coli DH5α as a host, amplifying a cloned formamidase gene and a cloned phosphite dehydrogenase gene into a fusion gene, ligating the fusion gene to a multiple cloning site of a vector, transforming the obtained recombinant plasmid into the E. coli DH5α, extracting the plasmid and transforming into an expression strain, and performing induction culture to obtain a recombinant E. coli. The recombinant E. coli can express a fusion protein of formamidase and phosphite dehydrogenase.

The invention relates to a recombinant Escherichia coli expressing a fusion protein of formamidase and phosphite dehydrogenase, a construction method and use thereof. The invention includes adopting engineered E. coli DH5α as a host, amplifying a cloned formamidase gene and a cloned phosphite dehydrogenase gene into a fusion gene, ligating the fusion gene to a multiple cloning site of a vector, transforming the obtained recombinant plasmid into the E. coli DH5α, extracting the plasmid and transforming into an expression strain, and performing induction culture to obtain a recombinant E. coli. The recombinant E. coli can express a fusion protein of formamidase and phosphite dehydrogenase.