Artificial gene synthesis is a critically important technique in modern synthetic biology. The ability to easily customize and create DNA sequences based off of a single gene, gene clusters, or completely from scratch enables an enormous amount of versatility and a wide range of applications for synthetic DNA.

The necessity of long gene synthesis

Researchers often investigate the genetic causes of a particular target phenotype, frequently by examining the effects of site-directed mutations. Through artificial gene synthesis, inducing mutations, stretches, elongations, or other changes to a target gene sequence becomes substantially easier than before, allowing for efficient study of these mutant phenotypes.

In recent years, the importance of being able to analyze entire genomes or large collections of genes has become more and more pronounced. The necessity of having to look at mutations in the context of an organism’s genome means that being able to synthesize huge amounts of DNA with near-perfect fidelity is becoming increasingly relevant to many current experiments.

Gene Synthesis Scheme

Current solid-phase oligonucleotide synthesis technology can yield chemically synthetic gene constructs. However, it is generally limited to sequences of roughly 200 nucleotides in length. Another approach named Gibson assembly shares a similar drawback in that it is restricted to plasmids of around 10kb or less.

The need for technology that could handle synthesis of very large DNA sequences turned to yeast as the system of choice. Yeast can provide self-connection between multiple DNA fragments easily and rapidly without the application of polymerase or ligase. Synbio Technologies’s own proprietary Syno® platform, combined with the built-in homologous recombination technology of yeast, enables rapid and accurate assembly of various long DNA fragments and genomes of up to 200kb.

Synbio Technologies delivers more than 2 million base pairs of DNA sequences every month, all over the world. Any DNA sequence, even those possessing difficult characteristics such as high or low GC content, hairpin structures, or highly long/complex sequences, is able to be synthesized with 100% accuracy guaranteed. Synbio Technologies can provide large sale and low cost methods to accomplish gene synthesis including assembling long DNA segments at high accuracy and yield.

Gene Synthesis Related Services

• Codon Optimization
• Vector Construction
• Small Genome Synthesis
• Pathway Synthesis

How to order

Get a quote: online request submission form.

Submit gene/protein sequence: Gene synthesis online inquiry.

Or please email your detailed sequences and requirements directly to: service@synbio-tech.com, Tel.: +1 732-230-3003. Project managers of Synbio Tech. will contact you within 24 hours after we receive your request.

Gene synthesis is the technology of artificially synthesizing double-stranded DNA in vitro, and is a crucially important technique in molecular biology. Conventional gene synthesis generally involves assembling small segments of DNA and/or amplifying known samples of genetic material, and requires the raw nucleobases guanine, cytosine, adenine, thymine and uracil as starting materials.

GC-content (guanine-cytosine content) refers to the proportion of two nitrogenous bases, guanine and cytosine, in DNA and RNA molecules, which might be any domain of a gene, single gene, gene clusters, or even non-coding regions. Synthesis of high GC-content sequences can be troublesome due to issues with secondary structure, mispriming, or mis-annealing. GC-rich regions tend to facilitate base stacking, which makes them more stable than sequences with low GC-content. Additionally, secondary structures formed by high GC-content regions also tend to be stable and more resilient to denaturation. Sequences containing many guanine repeats can also generate complicated inter-strand folding due to hydrogen bonds between adjacent guanines.

GC-content is closely related to temperature optimization in gene synthesis. In PCR, primer GC ratio strongly influences the predicted annealing temperature of DNA templates. High GC-content tends to require a high melting temperature, which can result in mispriming or mis-annealing between the template and its complementary strand, leading to undesired and inaccurate gene products. Even when GC-rich sequences or G repeats are located in noncoding regions, the secondary strucutres formed can still affect factors in gene synthesis, such as melting temperature. Thus, an important part of gene synthesis is codon optimization, using synonymous codons to lower the GC-content of a sequence and lowering the melting temperature.

Synbio Technologies is proficient at accurately synthesizing error-free DNA constructs meeting customers’ requests, and is capable of assembling multi-kilobase plasmids or even entire genomes. Synbio Technologies can make gene synthesis perfect by handling even the most challenging synthesis requests, including genes or genomes with various complex sequences such as hairpin structures, high GC ratio, high AT percentage, multiple consecutive nucleotide sequences, and so on. The successful synthesis of GC-rich constructs is important to fully understand and study complex genes and even their non-coding segments with high conservation, which in turn poses profound significance in biomedical research.

Gene synthesis has revolutionized what was once thought to be impossible in the field of genetics. Synthesis of the first gene, a yeast tRNA, was achieved by Har Gobind Khorona and coworkers in 1972. Since then this technology has had long last impacts on genetic research and the biotechnology industry. Whether it is synthesizing small genes, ranging in lengths from a few hundred base pairs in length, to large genes, up to 100kb in length, gene synthesis has made recently daunting research aspirations achievable. Gene synthesis has a simple definition, the creation of synthetic genes within a laboratory setting, but an impactful influence on many scientific fields. One field in particular, genetic engineering, is often associated with gene synthesis. The association between these two topics is so prevalent because gene synthesis is a popular mechanism to achieve the genetic engineering sought out by researchers. Genetic engineering also has a wide range of application in many different scientific fields, many relying on gene synthesis.

Gene synthesis itself has a many applications, ranging from developing more effective vaccinations to creating large synthetic gene libraries. Synbio Technologies offers the ability to take a requested genetic sequence, in text format, and create the desired physical sequence with one hundred percent accuracy. This is done with our, patent pending, Syno^®2.0 Platform. The unique feature that this platform offers is the capability of synthesizing a genetic sequence that may not be preexisting within any organism. Previously, the sequence of interest was required to be existing in order to be extracted and later cloned. The Syno^®2.0 Platform allows us to generate the user specified sequence exactly, confirmed with Sanger sequencing, with a both time and cost effective manner. This technology is the foundation for genetic engineering. Genetic engineering is defined as the alteration of an organism’s genome using biotechnology.

This is accomplished through the incorporation of a genetic sequence that is foreign to an organism into its genome. The unique connection between gene synthesis and genetic engineering is found within this step. Gene synthesis allows us to synthetically create the gene of interest, which is then incorporated into the organism’s genome. The gene of interest can have a wide range of functions, and are specific and tailor made to the interest of the researcher. Once the gene of interest is incorporated into the organism’s genome, research can be conducted to determine effects of the newly incorporated gene on the organism. This general outline has been applied to a wide range of applications in various fields of interest.

The applications of genetic engineering can be found in many fields of ranging from gene therapy, through the use of viral vectors, to agriculture. Gene synthesis and genetic engineering have been used to study the effects of a genetic knockout in a particular gene, a method that is very commonly used in genetic research. It is seen when studying the effects of gene amplification, causing an overexpression of a gene of interest. Genetic engineering is most commonly seen in agriculture through the creation of genetically modified crops to optimize the quality as well as quantity of certain crops. This process has been going on for thousands of years through selective breeding, but recent advances in technology have allowed researchers to implement groundbreaking innovations on basic crop production. These innovations help to combat environmental pressures, such as draught or blight, as well as decreasing pesticide use by genetic modification. These modifications, accomplished by genetic engineering, have a simple goal in mind: to increase the quality and quantity of these crops which are essential in everyday life. It is through genetic engineering and gene synthesis that these innovations are possible.

The main advantage that gene synthesis has when relating to genetic engineering is the accessibility and ease of synthetically creating your gene of interest. With Synbio Technology’s Syno^®Platform the process from desired sequence to genetically modified organism is easily achieved. This process starts with the input of the researcher’s requested sequence, up to 100kb in length. The requested sequences is created with one hundred percent accuracy and verified with Sanger sequencing. This step is done with a both time and accuracy efficient pipeline. The desired sequence will then be shipped to your location within as few as five business days. In addition to the time and accuracy efficiency Synbio Technologies offers an extremely cost effective approach, with prices starting at $0.25 per base pair for gene synthesis. The combination of competitive pricing, time efficiency, and accuracy have lead Synbio Technologies to be one of the leading biotechnology companies, especially within the realm gene synthesis and its relationship with genetic engineering.

Recently, gene synthesis has become an extremely popular and effective method to conduct genetic research. This rise in popularity stems from the ease of use as well as the effective results. In short, gene synthesis can be described as the creation of a synthetic genetic sequence specified by the user. Recently, the genetic sequence of interest must have been present within an organism to be extracted and later studied. Gene synthesis allows researchers to bypass this step entirely, creating the sequence of interest and converting the sequence from text format to physical copy with ease. This accessibility has caused multiple companies to participate in this groundbreaking technology within the field of genetics. As the gene synthesis industry has grown drastically, companies have competed to become the best at synthesizing genes. At Synbio Technologies we pride ourselves as being one of the leading companies within this gene synthesis industry. This pride and confidence is mainly relying upon our, patent pending, Syno® Platforms. It is the three phases of this platform that separates us from any other company in the gene synthesis industry, allowing us to serve you with a high quality product, with an efficient time frame and competitive prices.

The first platform that Synbio Technologies offers, the Syno^®1.0 Platform, is an industrialized DNA chemical synthesis platform. Essentially, this platform specializes in the formation of oligos. An oligo can be described as a short fragment of DNA or RNA molecule. These oligos have a wide range of functions in various fields of genetic research including forensics, and genetic testing. In order to accomplish the output of high quality oligos, Synbio Technologies has four steps within this mechanism. First, the DNA sequence of interest is analyzed and designed. Once this complete, CG-base oligo synthesis is conducted. This step includes industry leading and manufacturing process which allows for the highest quality oligos. In order to increase the quality, the oligos are then purified using the proprietary purification process. Finally, there is then an additional step of quality assessment using MS analysis to verify the created oligos are what the user requested. The Syno® 1.0 Platform has a wide range of applications using this four part mechanism. Some applications include generating native DNA sequences, creating de novo DNA sequences, and constructing degenerated oligo libraries and so on. All of these processes have been utilized and proven with the use of oligo synthesis. This is just the first step in the process, which has allowed Synbio Technologies to become one of the premier companies in the gene synthesis industry. The remaining two platforms are what really separate us from the competition.

The Syno^®2.0 Platform focuses on PCR based gene synthesis and is an aspect that Synbio Technologies prides itself as being one of the leaders in the biotechnology and gene synthesis industries. Synbio Technologies is capable of synthesizing a wide range of gene lengths up to and including 100kb. The process as to how these genes are synthesized are constantly verified for accuracy as well as quality. The resulting synthesized gene product is guaranteed to be one hundred percent identity to the user specified requested sequence. The resulting synthesized gene can then be applied in various types of genetic research. One major application that differs from many others is the ability to generate de novo DNA sequences. This aspect has revolutionized genetic research since the early 1970s, increasing in quality and effectiveness along the way. The creation of these de novo DNA sequences has allowed researchers to develop more effective vaccinations, generate variant libraries, and improve the features of protein. It is the basis of so many crucial fields of genetic research. At Synbio Technologies we offer the highest quality gene synthesis product with the most competitive prices in the gene synthesis industry. We pride ourselves with the accuracy of our product which we are able to provide our users.

The third and final platform, the Syno^®3.0 Platform, specializes in oligo pool synthesis. This platform is described as a chip-based next generation DNA synthesis platform. Essentially, this platform allows us to generate the highest quality DNA fragments, de novo synthesized genes, small genomes, and variant libraries. In addition to this, the high quality output of these various genetic materials is provided to the user at the lowest prices in the gene synthesis industry. The process that this platform follows is very similar to that of the Syno^®1.0 Platform with some small and necessary alterations. First the proprietary DNA sequences are designed, analyzed and optimized. Once this is complete, the chip-based oligo pool synthesis process begins. This step utilizes the, industry leading, oligo-pool assembly and manipulation process. After that, there is proprietary error correction, allowing for a quality control and improving the resulting output. Then large fragment assembly, up to 200kb, is conducted. Once this is finished, the final step is to run a quality control including next generation sequencing. This final step allows for the resulting sequence to be verified and one hundred percent accurate. Overall, the applications of this platform are similar to that of the previous Syno^®1.0 and 2.0 Platforms. The user can generate native and de novo DNA sequences, build genes, chassis, operons, pathways and small genomes, as well as generate DNA variant libraries. In addition to these features, this platform offers a unique aspect which allows the process to be scalable. This means that it will take the same amount of time to produce 20,000 genes as it would to produce 20 genes, making the Syno^®3.0 Platform more effective. These features are what make the Syno^®3.0 Platform extremely valuable to many users.

Gene synthesis is a fundamental technique in synthetic biology, allowing for the creation of unique synthetic genes that would be difficult or impossible to obtain naturally. In order to express a synthetic gene in an efficient and controlled way, expression vectors are often used to introduce the gene into a cell, after which the cell’s own gene expression system is used to synthesize the target protein.

Common Gene Synthesis Vector Types

There are three main categories of carriers that constitute vector systems in gene synthesis engineering: plasmids, bacteriophages and viruses. One of the most widely used expression vectors is that of Escherichia coli due to its ease of growth, rapid reproduction, and ease of transformation with exogenous DNA. Others include integration vectors, bacteriophage vector promotion, or shuttle vectors in S. cerevisiae.

Occasionally, a series of pGEX vectors (-1T, -2T, -3T), GST (glutathione S-trasferase) system, pEZZ18 are utilized for specific expression systems. Several variants transformed from pBluescript Ⅱ SK(+), together with pUC18, pUC19, pUC57, are also selected for certain projects.

Synbio Technologies are able to synthesize a large variety of target DNA and clone them into any vector as requested, regardless of the specifications of the project or the approach required. We guarantee a highly accurate and high-yield product at a low cost and with rapid turnaround.

Gene Synthesis Vector Design Application

Synbio Technologies provides two main approaches for amplifying DNA sequences via PCR cloning and subcloning technology. For DNA or amino acid sequences provided by customers, Synbio Technologies will synthesize error-free DNA de novo by request. Our proprietary Syno^®gene synthesis platform can perfectly achieve enzymatic assembly through limited short DNA fragments. Even genes with challenging characteristics such as repetitive sequences, complex secondary structure, and high (>80%) or low (20%) GC content as well as long polypyrimidine runs, can be synthesized correctly.

Additionally, our professional Syno^®Codon software can compute and optimize codon selection for original sequences or specific vectors. Any synthetic vector meeting our requirements can be quickly and accurately delivered at an economical price. Synbio Technologies can insert any target gene into any site of any vector, and can guarantee a high-quality, fully accurate final product for gene synthesis.

Introduction

Over the last few decades, gene synthesis and assembly technology have developed rapidly. Since the 1960s, our capability to synthesize genes has skyrocketed from less than 100bp to more than 1,000,000bp. Gene synthesis methods and applications have a profound impact in metabolic engineering, genetic network design, and vaccine design. However, existing gene synthesis methods still have their fair share of drawbacks and inconsistencies, such as low yield, high error rate, and lack of scalability both on the size of the project and the size of the gene.

Gene Synthesis Methods

Gene synthesis methods are not able to replace each other, and each occupies its own niche depending on the requirements of the project. The following is a brief overview of several common gene synthesis methods:

• Solid-phase synthesis

The traditional oligonucleotide synthesis uses a small volume of solution processed in a column full of chemicals. The oligonucleotides are synthesized by attaching nucleotide residues stepwise to the end of the chain, one-by-one. The addition of each oligonucleotide consists of four steps: de-blocking, coupling, capping, and oxidation. The integrity of the sequence and the productivity of the synthesis are hindered for products longer than 200bp, and thus this method is generally limited by DNA sequence length. The primary advantage of this method is its high accuracy, compensating for its high expense and low output.

• Chip-based DNA synthesis

As the name implies, Chip-based synthesis utilizes microarray chips utilizing a series of electrochemical techniques. Different kinds of oligonucleotides are able to be synthesized in different specific parts of the chips, called assembly subpools. Following this piecewise synthesis, gene fragments in subpools are amplified and then aggregated and assembled into the finished product. Chip-based DNA synthesis is cheaper than solid-phase synthesis and can yield a larger amount of the target gene, but its accuracy suffers in comparison.

• PCR based enzyme synthesis

PCR-based enzyme synthesis generates gene fragments through a variety of cell systems. Using the Yeast system as an example, by using different incision enzymes and label markers, different kind of genes can be added to Yeast chromosomes. Due to the nature of gene insertion the target gene could have no limit to its length as long as the chromosomes can accommodate. This method performs well in synthesizing large gene fragments, and with the help of the cell systems the accuracy of the gene sequence is guaranteed.

Gene synthesis applications

Synthetic genes have wide implications on a variety of fields, ranging from genetic circuits to metabolic improvement to many more. As our technology advances and our understanding of genetics continues to develop, scientists could modify and design genes, The scientists have already synthesized and assembled a gene fragment of over 100kbp. When inserted into a host bacterium lacking its own genetic material, the bacterium was able to successfully produce new cells.

Gene synthesis methods can even be used to construct new metabolic systems in living cells. For example, Jay D. Keasling constructed a new metabolic system in E. coli and S. cerevisiae in the mid2000s to produce artemisinin. An important component in some anti-malarial drugs, Keasling’s construct reduced the cost of artemisinin production by tenfold, showing that gene synthesis has a vast amount of potential in medicine and countless other fields.

Gene synthesis was first successfully conducted in 1972 on a yeast tRNA by Har Gobind Khrona and associates.Since 1972, the technology and mechanisms of gene synthesis have flourished in order to successfully obtain the genetic sequence specific to a wide range of research topics.Gene synthesis offers many major advantages over the traditional gene cloning methods: molecular cloning and polymerase chain reaction (PCR).These two traditional methods have two major restrictions,the first is that these methods are only capable of gene amplification and cloning, as opposed to gene synthesis.The other restriction molecular cloning and polymerase chain reaction have is that the preexisting genetic sequence of interest must be obtained and then amplified.The important advantage gene synthesis offers is that the genetic code you wish to study does not need to be physically obtained in order to be amplified.The technology currently allows us to synthesize “de novo” sequences.This means that a sequence of interest,whether it is viral DNA or a cancer cell’s mutation in a specific gene, can be synthesized without the physical copy of the gene being present.Without the physical copy of the gene of interest being needed,the necessary amount of time and money is drastically reduced when compared with traditional methods of cloning, amplification,targeted mutagenesis,and gene target knockouts.Due to this advantage gene synthesis has been becoming more and more popular in various fields of research.

Due to the ease of use and cost effective mechanism, gene synthesis has been used countless amounts of times in varying fields of scientific research. Gene synthesis has most notably and recently been used to conduct viral research in order to produce safer, more effective DNA vaccines. It has also been used to better understand mechanisms for cancer cell metabolic regulation. Gene synthesis is currently being used for targeted gene therapy, and other topics of interest that seemed almost impossible 30 years ago. In addition to the research that has already been conducted, gene synthesis offers a myriad of possibilities of application ranging from gene regulation to better understanding evolution and antibiotic resistance. Gene synthesis makes it possible to build variant libraries, genes, operons, increase the function of proteins, and even test all the orthologs of a particular gene.

The advantages gene synthesis offers are seemingly endless.The main advantage that gene synthesis has to offer is the ease of synthetically manipulating or cloning a gene of interest with or without the physical copy of the gene itself.Synbio Technologies offers, through our Syno^®DNA Platform, an extremely accurate, time efficient, and cost effective mechanism to analyze your sequence of interest and successfully synthesize the sequence.All sequences constructed by Synbio Technologies are verified using Sanger sequencing, and are guaranteed to be 100 percent identical to your sequence of interest.In addition to the high quality synthesized sequences, Synbio Technologies offers competitive prices on gene synthesis for a wide range of gene lengths up to 100 kb.With the competitive prices and accuracy, on top of the additional money and resources needed for PCR and molecular cloning, it is clear that gene synthesis is the more effective mechanism to utilize when conducting certain types of genetic research.