Gene synthesis is an efficient and cost-effective alternative to molecular cloning for custom gene production. When compared to PCR cloning, GENEWIZ gene synthesis offers faster turnaround time for a comparable price. For a comparison of GENEWIZ custom gene synthesis services versus traditional molecular cloning, please click here.

Single-stranded oligonucleotides are designed, synthesized, and assembled using different protocols depending on sequence characteristics. Subsequently, the assembled full-length gene is cloned into the pUC57 vector; the insert is verified by Sanger DNA sequencing and restriction digestion.

GENEWIZ will synthesize cloned gene sequences ranging from 1 bp to 50+ kb, using short DNA oligos as building blocks. For standard priority gene synthesis, products delivered as purified PCR fragments are limited to 200 bp to 1.2 kb.

GENEWIZ has a proven track record of synthesizing difficult templates, such as GC-rich sequences and sequences with repeats, using proprietary technologies. With our bioinformatics platform, GENEWIZ optimizes your gene for synthesis. In some very rare instances, extreme sequence content may be refractory to available sequence verification techniques. GENEWIZ carefully reviews every sequence to estimate difficulty and anticipate project challenges, and customizes every project quotation to include an estimated turnaround time and pricing information based on this analysis.

*Applicable to non-complex genes delivered in a standard pUC57 vector.

GENEWIZ provides a custom quote for every gene synthesis project. For standard priority, typical gene sequences (non-repetitive, average GC-content) that do not require custom subcloning, no additional fees apply. For genes with expedited handling, difficult sequences, large inserts, or for projects with custom subcloning requirements, any relevant fees will be indicated in your custom gene synthesis quote.

GENEWIZ can deliver codon-optimized genes, designed to meet your exact research specifications.

GENEWIZ codon optimization tool has the ability to optimize multiple critical parameters to stabilize DNA fragments and improve gene expression efficiency. Parameters include but are not limited to:

• Codon Usage Bias;
• GC content;
• mRNA Secondary Structure;
• Custom Desired Patterns;
• Custom Undesired Patterns;
• Repeat Sequences (direct repeat, inverted repeat, and dyad repeat);
• Restriction Enzyme Recognition Sites (deletion or insertion).

The assembled full-length gene is cloned into the pUC57 vector, and verified with Sanger DNA sequencing on at least one strand and restriction digestion.For standard priority gene synthesis, products between 200 bp and 1.2 kb may also be delivered as a purified PCR fragment, and are verified by Sanger DNA sequencing on both strands.

pUC57 contains several common restriction sites. Standard cloning into pUC57 typically occurs via EcoRV, however the exact restriction site(s) used for each sequence may vary; specific restriction sites within the MCS may be maintained or excluded upon request.

GENEWIZ offers both ampicillin and kanamycin marked version of pUC57, at no charge.

Customers must supply vectors other than pUC57. If you need to clone your synthetic gene into a custom vector, please send GENEWIZ the vector DNA and pertinent information (i.e. vector size, cloning site, copy number, etc).

GENEWIZ employs quality controls throughout the duration of all gene synthesis projects to ensure accurate synthesis, starting with inclusion of your exact sequence for synthesis within your custom gene synthesis quote. This enables customers to review and confirm the sequence prior to project initiation.

Once beginning your project, GENEWIZ conducts sequence verification during the gene assembly process, verifying that any subassemblies contain the correct sequence. Subsequently, the final plasmid DNA is clonally isolated and confirmed to match your sequence by Sanger DNA sequence analysis to at least single-strand depth. Purified PCR fragments are sequence verified on both strands, although verification at the fragment ends is limited to single strand depth. Upon completion of your synthetic gene, GENEWIZ delivers an electronic data package in parallel with your construct, containing the original sequencing data files from construct verification for your independent review.

Final deliverables for GENEWIZ gene synthesis projects include:

• Certificate of Analysis (COA), including a gel image of a restriction digest test (cloned vector products, only) or purified PCR fragment;
• Alignment of sequencing data with the complete insert sequence;
• The complete vector sequence, if applicable.

Standard deliverables for GENEWIZ gene synthesis projects include:

• 2-5 µg of lyophilized plasmid containing your desired synthetic construct;
• Certificate of Analysis (COA), including restriction digest;
• Sequence trace data with alignment;
• Sequence of synthetic gene alone and in the designated vector.

Standard deliverables for GENEWIZ purified PCR fragment synthesis projects include:

• 1-2 µg of lyophilized plasmid containing your desired synthetic construct;
• Certificate of Analysis (COA), including gel image;
• Sequence trace data with alignment;
• Sequence of synthetic gene alone.

GENEWIZ sample tubes contain 2-5 µg lyophilized plasmid DNA, or 1-2 µg purified PCR fragment DNA for standard synthesis projects. For resuspension, we recommend:

• Spin the tube briefly to ensure that the contents are on the bottom of the tube;
• Resuspend the sample in a volume appropriate for your needs:
  • For example, 20-50 µL for ~0.25 to ~0.1 µg/µL of sterile high purity water or TE;
  • The resuspension solution is dependent on downstream applications - TE may interfere with various enzyme manipulations;
• Vortex briefly, let the DNA tube sit for 2-10 minutes on ice, then vortex again.

GENEWIZ is fully aware of all risks and benefits associated with gene synthesis research. With the knowledge that gene synthesis technologies have the ability to enable de novo reconstruction of dangerous pathogens, GENEWIZ employs rigorous quality control policies and procedures to safeguard against abuse of the genes we synthesize. GENEWIZ actively monitors the Screening Framework Guidance for Synthetic Double-Stranded DNA Providers, drafted by U.S. Department of Health and Human Services (HHS), as well as the Select Agents and Toxins list put forth by the U.S. Government.

At GENEWIZ, protection of our customers' intellectual property is a top priority. Proprietary information is always under client ownership, and GENEWIZ places the utmost importance on confidentiality and privacy. Comprehensive protective measures enable GENEWIZ to earn and maintain the trust and confidence of all customers, collaborators, and partners. For more information about the GENEWIZ Confidentiality Policy, please click here.

To request a gene synthesis quote, please click here to complete the simple online quote request form. GENEWIZ will contact you within one business day to discuss your project in detail.

For large orders, please email your sequences and project details directly to genesynthesis@genewiz.com. GENEWIZ Project Management will contact you within one business day to discuss your project in detail.

To request a TurboGENE^TM quote, please click here to complete the simple online quote request form. GENEWIZ will contact you within one business day to discuss your project in detail.

GENEWIZ Project Management is composed of specialized experts dedicated to your project's success. Project Management is available to assist with any questions throughout the duration of your gene synthesis project. In addition, GENEWIZ Project Management will provide milestone updates at each stage of your gene synthesis process, keeping you well-informed throughout all stages of research.

To contact GENEWIZ Project Management, please click here.