GeneArt™ Seamless Cloning and Assembly Enzyme Mix enables the simultaneous and directional cloning of 1 to 4 PCR fragments, consisting of any sequence, into any linearized vector, in a single 30-minute room temperature reaction. GeneArt™ Seamless enzyme mix is the economical choice for creating constructs up to 13 kb with the option for high-throughput assembly.

• Efficiency: pre-cloning option for large fragments for increased cloning efficiency
• Speed and Ease: clone DNA fragments, with sequence of your choice, in a single vector (up to 13 kb); no restriction digestion, ligation, or recombination sites required
• Free Tools: design of your final construct and DNA oligos in silico using our free web-based tool that takes you step-by-step through your project
• Vector Flexibility: use our linear vector or a vector of your choice

For the cloning of pre-existing DNA elements too long to be amplified by PCR and for final molecules up to 40 Kb, please consider the GeneArt™ Seamless PLUS Cloning and Assembly Kit (1–4 DNA fragments only); and for final molecules larger than 110 Kb, please consider the GeneArt™ High-Order Genetic Assembly System.

Simple and Fast Clone Creation
GeneArt™ Seamless Cloning is a simple, two step process, consisting of in vitro assembly followed by transformation. The enzyme mix uses a proprietary enzyme/buffer mix to assemble DNA fragments without extra sequences or scars ('seamless'). Terminal end homology, if needed, is easily incorporated by PCR amplification with custom DNA oligos engineered using our free web tool.

Cloning Efficiency, Flexibility, and Precision
Cloning success is independent of the insert sequence and vector type, allowing you to design and add nearly any desired sequence, or combination of sequences, to any plasmid as long as it can be linearized by either restriction enzyme digestion or PCR. The circularized clones obtained from the reaction contain only the sequence of your original vector, inserts, and designated homologies, with no extraneous nucleotide insertions.

in silico Cloning Design Support
A key step in GeneArt™ Seamless Cloning is the correct design of fragments and oligos with the appropriate homology and spacing to help ensure successful assembly of your clone. To simplify and speed the design process, we provide a free online tool, the GeneArt™ Design Tool for Seamless or High-Order Assembly and Mutagenesis, to help you design your experiment in silico. The tool guides the user through construct design, including fragment import, order, and orientation; checks for areas of homology and potential design issues; creates primers for pre-cloning or incorporating end homology between fragments, if needed; and generates final construct map files and sequence in Genbank format for download or import into Vector NTI™ software.

Applications
The GeneArt™ Seamless Cloning and Assembly Enzyme Mix is designed to empower cloning and DNA assembly in a wide range of molecular biology and synthetic biology applications, among others. The product allows for the creation of modular expression vectors, with interchangeable parts, and can be used to perform a variety of tasks that would otherwise involve multiple steps. Use the mix to construct fusion proteins; delete, replace, or add DNA elements such as restriction sites in an existing vector; and carry out many other techniques that require manipulation of genetic sequences.

For Research Use Only. Not intended for human or animal therapeutic or diagnostic use.