E. coli for recombinant vector selection and growth.

•Quick and easy—Clone up to 4 DNA fragments (containing sequences of your choice) simultaneously in a single vector (up to 13 Kb); no restriction enzyme digestion, Joining or Recombination Sites
•Precise and Efficient—Designed to enable you to clone the fragments you need, in the direction you need, at the site you need, and get up to 90% Correct cloning and no extra sequence left behind
•Vector Flexibility—Use our linear vectors or a vector of your choice
•Free Tools— Design DNA oligonucleotides and more
•Multiple Applications—Simplify a variety of synthetic biology and molecular biology techniques by quickly combining, adding, deleting, or exchanging DNA fragments

For cloning of more than 4 DNA fragments , the final molecule is larger than 110 Kb, or the pre-existing DNA element used is too long to be amplified by PCR; please consider using GeneArt™ Advanced Genetic Assembly System (Cat. No. A13285).

Easy and fast clone creation
GeneArt™ Seamless Cloning is a simple two-step process that includes a tube-based assembly reaction followed by conversion to One Shot™ Chemistry The process of TOP10 E. coli. The kit leverages the properties of a proprietary enzyme mixture to assemble DNA fragments with shared end homology without leaving any additional sequence or traces (seamless). A portion of the assembly reaction is then transformed into the provided competent TOP10 cells, thereby generating clones ready for analysis the next day. The required 15 base pairs of end homology can be easily engineered by PCR amplification using custom DNA oligonucleotides.

Efficient, flexible and precise cloning
When using the GeneArt™ Seamless Cloning and Assembly Kit, the main factor affecting cloning efficiency is the DNA element size (100 bp to 5 Kb), total final molecular size (≤ 13 Kb), and fragment quality and specificity. The ends of the PCR fragments (A overhangs or blunt ends) do not affect cloning efficiency.

Typical cloning efficiencies for different numbers of fragments cloned into pUC19L are as follows:
• 90% for a single 5 Kb DNA element
• 70% for 4 fragments (1 Kb each)
• 40% for 4 DNA fragments (2 Kb each)

Cloning success is independent of insert sequence and vector type, you can design and add almost any sequence or sequence combination you want into any plasmid, provided the plasmid can be linearized by restriction endonuclease digestion or PCR. The circular clone obtained from the reaction contains only your original vector sequence, insert, and specified homology, with no extraneous nucleotide insertions.

In silicoClone design support
A critical step in GeneArt™ seamless cloning is the creation of fragments and oligos with appropriate homology and spacing. Correct design of nucleotides to help ensure successful assembly of clones. To simplify and speed up the design process, we offer a free online tool to help you through Computer simulationDesign experiments. The tool checks experimental design for compatibility with product specifications, designs DNA oligonucleotides with end homology for PCR amplification of different elements for cloning, and shows the user a graphical representation of the vector and its interaction with Vector NTI™ Downloadable annotated sequences in software-compatible GenBank format.

Applications
GeneArt™ Seamless Cloning and Assembly Kits are designed for cloning and DNA assembly experiments in a variety of molecular biology and synthetic biology applications. The product can be used to create modular expression vectors with interchangeable parts and can be used to perform a variety of tasks that might otherwise involve multiple steps. Use kits to simplify: constructing fusion proteins, deleting, replacing, or adding DNA elements (such as restriction sites) to existing vectors, and many other techniques that require manipulation of genetic sequences.