Custom Gene Synthesis Services

The sequence-verification method depends on the manufacturing workflow. Ansa enzymatically synthesized clonal genes are verified using PacBio HiFi long-read sequencing, while IDT chemically synthesized genes are verified using short-read next-generation sequencing (NGS).

• PacBio HiFi: Long reads can span the complete plasmid end to end, helping confirm sequence context across long, repetitive, or structurally complex regions. This method may generate fewer individual reads than short-read NGS, but each read covers more of the construct.
• Short-read NGS: Many overlapping reads provide deep coverage and are well suited to routine constructs. Because the DNA is fragmented before sequencing, coverage can vary, and repetitive, highly structured, or extreme-GC regions may be more difficult to align.

Both methods verify the final construct against the ordered sequence. The quality-control method is matched to the synthesis workflow and construct characteristics.

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We store onboarded custom vectors for two years after the most recent order using that vector, at no additional fee.

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We are open to reviewing your vector if it does not meet the above criteria. Please contact your regional scientific applications support team with details of your vector, providing the sequence in FASTA format, if possible.

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Available vector options depend on the sequence length, complexity, and synthesis workflow. Some complex or XL designs may be limited to compatible catalog vectors.

The ordering portal will show the available options after your sequence is screened. If you need a different vector or cannot complete the order online, please contact us. At this time XL genes are only in the pStable vector.

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We’re actively expanding the range of lengths and complexities available for online ordering. To ensure a high-quality customer experience, these capabilities are being rolled out in phases.

We will notify customers as new options become available. If you’re interested in ordering higher-complexity genes or constructs up to 50 kb today, please contact us for early access.

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Ansa-manufactured genes are delivered with a guaranteed minimum yield of 250 ng and typically ship at approximately 1.5 µg (non-normalized).

DNA quantity is measured using a fluorescence-based method (Qubit®, Thermo Fisher Scientific), which may report a lower concentration than absorbance-based methods such as NanoDrop® (Thermo Fisher Scientific). If you have questions about yield or quantification, please contact us.

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Order custom genes or Ansa genes through our online Gene Synthesis ordering portal. Enter your sequence and project requirements, and the ordering experience will present the available synthesis and vector options based on length, complexity, and feasibility.

Start an order. If your design cannot be ordered online or you need help placing the order, please contact us.

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IDT Genes and Mini Gene™ sequences are delivered as double-stranded DNA, cloned into your provided custom vector.

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Please provide the 50 bases of sequence flanking the insertion site you wish us to use when submitting a new request through the Custom Vector intake tool. The vector's single fixed insertion site is defined by these flanking sequences. IDTs proprietary assembly method requires no restriction sites and can remove existing sequences from a vector. 

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Simple plasmid preps using a spin column or alkaline lysis method of purification are acceptable. Vectors greater than 8 kilobase pairs in size, those which require specialized cell lines or growth conditions for propagation, or those with exceptionally low yields may incur additional costs and/or turnaround time for gene production after validation.

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• An E. coli origin of replication
• Kanamycin, Ampicillin, or Chloramphenicol resistance marker
• 1 µg of circular plasmid

A minimum of 50 bases of sequence flanking the insertion site you would like us to use for the assembly of your gene inserts into the vector. Therefore, you do not need to specify restriction sites for assembly.

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Synthesis time depends on the sequence length, complexity, selected options, and synthesis workflow. The expected ship date for each gene is shown in your order table after sequence screening.

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IDT offers custom gene synthesis for sequences from 25 bp to more than 5 kb. Expanded Ansa enzymatic DNA synthesis provides access to more complex DNA and sequences from 100 bp up to 50 kb.

Submit your sequence through the online ordering portal to see available options. If your design cannot be ordered online, please contact us.

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We have removed the multiple cloning site and most common restriction sites from our standard vectors. This allows you to add convenient restriction sites to your sequence for subcloning into your destination vector.

Add the required restriction sites when placing your order. For more information about our standard vectors, please contact us.

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By not having a set multiple cloning site in our vectors, customers are able to choose the optimal restriction site set required to subclone into their choice of destination vector. You should add the restriction sites to your sequences when placing your order.

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Yes. A customer-supplied vector must be validated before it can be used for gene synthesis.  Please provide:

• The vector sequence, preferably in FASTA format
• At least 50 bases flanking the requested insertion site
• 1 µg of circular plasmid
• An E. coli origin of replication
• A kanamycin, ampicillin, or chloramphenicol resistance marker

Standard spin-column or alkaline-lysis plasmid preparations are acceptable. Vectors larger than 8 kb, vectors requiring specialized cell lines or growth conditions, or exceptionally low-yield vectors may require additional cost or turnaround time. A minimal onboarding fee applies; after successful onboarding, there is no additional fee for using the vector. If your vector does not meet these requirements, please contact your regional scientific applications support team for review.

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IDT ships custom genes dry unless otherwise requested. While dry, they should be stable for up to 2 years. After resuspension in  high-quality, molecular-grade water or buffer, pH 7.5–8.0, store the DNA at –20°C and, if needed, aliquot to avoid more than 2 or 3 freeze-thaw cycles.

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Plasmid DNA is fragmented and converted into a sequencing library before NGS analysis. Although adapter insertion is partly random, sequence structure, GC content, repetitive regions, and other complexities can affect local read depth and read quality.

Even clonally pure samples rarely show 100% identity in every NGS read. Sequencing and analysis errors are typically about 0.1% to 0.01% and may be locally elevated in difficult regions. Trace misalignment and small subpopulations of variants arising during E. coli propagation can also contribute.

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Even with clonally pure samples, NGS traces rarely show 100% sequence identity. This is because errors do occur during sequencing reactions and the subsequent data analysis.

These error rates are typically between 0.1% and 0.01%, but they can be locally elevated in regions of extreme GC content or secondary structure. Misalignment of the traces can also occur, particularly in highly repetitive sequences.

Finally, small subpopulations of sequence variants can arise during propagation of E. coli.

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Synthetic genes up to 5 kb are generally delivered as double-stranded DNA cloned into a best-fit IDT vector, unless another available vector option is selected. Ampicillin- and kanamycin-selection options are available.

The documentation supplied with your product identifies the vector, vector sequence, and insertion site. View IDT cloning vectors and sequence information.

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