FAQ DNA2.0 FAQ
Answers to your most pressing questions.

Have a question not answered below? Please contact us at +1 877 362 8646 or info@DNA20.com

Are you in the lab to clone genes or do science?
Gene synthesis saves you time and money compared to conventional cloning, allowing you to focus on your research.
Create sequences that do not exist in nature or cannot be easily made by cloning. Redesign your sequences to achieve high protein expression, easy manipulation, no promoter leakiness and convenient protein tags.
Single-stranded oligonucleotides are assembled using a variety of proprietary protocols depending on the sequence characteristics. The assembled oligonucleotides are then cloned and the complete double strand sequence verified. DNA2.0 routinely manufactures genes from 200bp to 60kb.
Yes, that’s what we do.
Two parts of a protein? Sure.
Only know the amino acid sequence? Yes, we can do that too.
We don’t need a template or a clone, it’s fully synthetic. All we need is the nucleotide or amino acid sequence.
We can synthesize gene sequences of any size.
Most genes range from 200bp to 60kb, with the greatest gene length continually increasing.
DNA2.0 is the industry leader at synthesizing long and/or difficult sequences. Use our GenomeGPS® system to design and synthesize entire genomes.
Want to set a new length record? Give us a call.
There is no upper limit. We have made synthetic DNA fragments of over 230kb. Create a novel protein, vector or even strain de novo.
DNA2.0 is the industry leader at synthesizing long and/or difficult sequences. Want to set a new length record? Give us a call.
Yes. DNA2.0 proprietary and patented technologies enable us to synthesize difficult sequences.
Rest assured that all your genes are made in sunny California, 100 percent accurate and intellectual property compliant. DNA2.0 never offshores or outsources any gene synthesis to third parties. All orders are shipped Overnight from California to arrive Worldwide in 48 hours or less.
Yes. DNA2.0 understands the value of your intellectual property and provides airtight IP protection. DNA2.0 synthesizes all orders at our state-of-the-art facility in Menlo Park, California, never outsourcing to countries with lax IP protection.
There are over 600 independent, peer-reviewed, published papers using DNA2.0 genes. They appear in many different journals including Cell, Nature, Science and PNAS.
These references can be found here.
DNA2.0’s gene synthesis service includes:
  • Optimization with our patented GeneGPS® algorithms
  • Synthesis of the DNA
  • Cloning into your chosen vector
  • 2-5 µg purified, plasmid DNA with cloned insert unless otherwise specified. Product may be delivered in tube or plate format (large orders primarily). International shipments are shipped on filters.
  • Quality Assurance Certificate.
  • Double stranded DNA sequencing trace files posted via your secure, online account.
  • Plasmid map plus FASTA files of the full insert and vector sequence posted via your secure, online account.

Easily design on your own

Download the FREE Gene Designer software. Gene Designer enables you to put your imagination to work guiding you through every step, from inspiration to synthesized gene with patented drag-and-drop sequence functionality to design genes de novo.

Let us help you

Please contact us for assistance with any synthetic gene design. We have years of experience designing and constructing synthetic genes with many different features, for use in various organisms. Most of our design assistance is free of charge.
Yes. DNA2.0 synthetic genes are delivered cloned into your choice of vector.
Virtually any vector you want.
Choose from one of our many DNA2.0 cloning vectors or expression vectors (Bacterial, Mammalian, Yeast).
Or, choose Custom Cloning and we’ll clone your gene into any vector you provide us.
Yes. With DNA2.0 Custom Cloning, your gene can be delivered in your vector for an additional charge. Please provide 5µg of purified plasmid DNA, plasmid sequence, selection information, and detailed information of the required cloning sites.
There are none. You can add any tag to your gene sequence that meets your individual research needs.
Yes. Simply include these in your sequence or note on the quote or order form. You can also note any sites you wish us to avoid.
You should include restriction sites in your gene design if you wish to remove your gene from the vector with restriction enzymes. Please refer to the vector information pages for specific information regarding restriction sites for your selected vector.
Yes. We can resend any gene. Cost and timing will depend upon individual circumstances.
Yes. DNA2.0 offers a complete selection of mutations and splice variants, complex block variants and SNPs.
Variants are always 100 percent sequence verified and individually cloned into the same vector as the primary gene. Gene Variants are often available at a significantly discounted rate compared with the primary gene, and can be ordered at any time.
Learn more about Variant Synthesis.
Yes. DNA2.0 has developed seven dynamic library options to enable you to focus your time directly on your research while quickly achieving significant results.
Modular Libraries (BioBricks) and Antibody Libraries are the most commonly utilized in research. Combinatorial Site Libraries and Site Scanning Libraries are used to sample a designed set of sequence variations, and screen to identify improved or novel properties.

Yes. You can choose to have your sequence optimized with our GeneGPS®. Using proteins with the highest levels of expression saves you time and money, while getting you quickly to meaningful results.
Only if you want more protein.
A gene that has been designed using optimized with our GeneGPS® optimization algorithms will yield between 10- and 100-fold more protein. This increase saves you months of time on painstaking optimization studies, allows much smaller culture volumes, and provides the necessary material for your research.
No. Breakthrough research by DNA2.0, funded by the NSF, has identified the design principles by which codons are used to maximize protein expression. The surprising results were published in PLoS ONE. Based on the results, DNA2.0 has created GeneGPS®, a set of design algorithms that offer the researcher 10-100 times greater expression yields than genes designed using the algorithms of our competitors.
Primarily because it does not work. With codon optimization you want the codon bias distribution for your gene to reflect the codons that confers expression during artificial induction. Research shows this is very different than using the most common codons (high CAI).
Research shows that high CAI does not actually maximize expression. With codon optimization you want the codon bias distribution for your gene to reflect the codons that confers expression during artificial induction.
Yes, enjoy:
Read the details of DNA2.0′s revolutionary gene optimization technology published in our peer-reviewed paper, “Design Parameters to control synthetic gene expression in Escherichia coli.” published in PLoS ONE (2009).

Read our review “Engineering Genes for Predictable Protein Expression” published in Protein Expression and Purification (2012). Request pdf.
There is no effect on solubility. If it’s soluble before, it will be soluble after optimization and synthesis. If it’s 30 percent soluble before, it will be approximately 30 percent soluble after optimization.
It often makes a big difference to the protein expression level. This is probably because codon optimization changes the DNA/mRNA sequence and removes intragenic regulatory elements as well as adjusting the codon bias. Remember, genes have often evolved to be tightly regulated, not for maximal expression.
Gene Designer does not contain the proprietary DNA2.0 GeneGPS® optimization algorithms. You can use Gene Designer for sequence design and preliminary optimization, but if you want maximal protein expression, you should send your sequence to DNA2.0 to be optimized.
No. If you want maximal expression in E. coli, yeast, or mammalian host systems, you should let us optimize your gene using our patented GeneGPS® optimization algorithms.

Just click on the “Get Quote|Order” button in the upper right corner on every webpage to go to our online quote request tool.
Or, Click Here to go to our online quote request tool.
The tool will quickly and easily guide you to:
  • Enter your DNA or Amino Acid sequence(s) of interest
  • Choose the vector (cloning or expression) for your final cloned sequences
  • Choose GeneGPS Optimization (if desired) and for what species
  • Select restriction sites, tags or motifs to include at 5’ and 3’ ends and/or to avoid internally
  • DNA Quantity (standard or large scale prep)
  • Select Synthesis speed (Standard, Priority or Rush)

Or, email us the above information. You can also send us a quote request using a Gene Designer or text (.txt) file in fasta format.
Yes. On the first step of the Gene Synthesis Quote/Order form, you can select the “Upload a file” tab.
You can upload your sequences from a Gene Designer file or a FASTA text file or an Excel template. Note, Word files do not import correctly, these must be saved as .txt files.
Yes. Consult with a synthetic biology specialist today at +1 877 362 8646 or info@DNA20.com to achieve your research goals quickly and affordably.
Yes. Our Ph.D-level support team are experts in the fields of protein expression, gene synthesis and the voodoo of cloning. DNA2.0 is your partner for accurate and innovative solutions.
All orders are shipped UPS Overnight from California to arrive Worldwide in 48 hours or less. US and Canadian customers can expect genes to be delivered the day after Ship date. European, Australian and Asian customers can expect 1-2 days for shipment to arrive.
Yes, with a few exceptions. We know how important it is for you to receive your synthetic genes as fast as possible. That is why we have the only turnaround time guarantee in the industry.

Gene Length Standard Delivery Priority Rush
Business days, guarantee is subject to restrictions (see below).
<1kb 10 days Inquire 5 days
1kb – 2kb 14 days Inquire 10 days
2kb – 3kb 18 days Inquire 12 days
>3kb Inquire Inquire Inquire
Follow synthesis status of your gene in real time directly in your online DNA2.0 account.

Guarantee: DNA2.0 will guarantee eligible genes (see Restrictions) to ship from DNA2.0 within the above turnaround times once an order has been approved. Eligible genes that fail to meet the business day turnaround time posted will be credited with half the gene cost on a DNA2.0 GeneCard for future orders.

Priority Service: Move your genes to the top of the queue at every step in the gene synthesis process. Not quite as fast as our signature RUSH service, but genes generally ship 20% faster than standard delivery. Actual delivery times are based on the individual gene and cloning characteristics.

Restrictions: DNA2.0′s turnaround guarantee is not valid for orders requiring custom cloning, low copy vectors, genes encoding high GC, stretches of homopolymers, extensive repeats or genes longer than 3kb. Turnaround times can be affected if a gene is toxic to E.coli at propagation.
You can follow the synthesis status in real time for each gene directly in your online DNA2.0 account.
We accept major credit cards (MasterCard, Visa, American Express), Purchase Orders or Wire Transfers (fee applies). We also offer prepaid GeneCards to expedite purchases and assist with your internal tracking.
$50.00 USD per shipment. Anywhere in the world.
Safety and biosecurity are a chief priority for DNA2.0.
We are a founding member of the International Gene Synthesis Consortium (IGSC). The consortium has established a Harmonized Screening Protocol to promote biosecurity and is actively working with authorities to devise the most effective biosecurity practices. DNA2.0 follows a strict biosecurity compliance code, screening all incoming orders against the IGSC list of select agents. This list is based on sequences derived from Center for Disease Control (CDC), US Department of Agriculture (USDA), and the Australia Group. We reserve the right to refuse any order.
DNA2.0 considers all account contact information, payment information and DNA/protein sequences submitted by customers as strictly confidential. Information is only disclosed to employees at DNA2.0 bound by confidentiality agreement on a need-to-know basis to enable the individual to perform their duties. Information is only used for the purpose for which it was disclosed. Confidentiality agreement is available upon request.

DNA2.0 may send e-mail newsletters or other types of directly related information to e-mail addresses corresponding to individuals who have opened an account with DNA2.0, have requested a quotation, have downloaded Gene Designer or have directly requested to be added to our mailing list. DNA2.0 will never rent, sell or otherwise make e-mail addresses, postal addresses or telephone numbers available to third parties. DNA2.0 does not purchase or rent e-mail lists from third parties. You can unsubscribe at any time for any reason quickly and easily by e-mailing us at communications@DNA20.com.
DNA2.0 ships directly to all corners of the world. We do not use distributors, so we can always speak directly with customers. We provide all technical support without information ever being lost on the way.

Find help on using Gene Designer Here.

DNA2.0 has created a line of products designated “IP-Free”. These products do not require any licensing, do not have royalty shares and do not have restrictions on downstream use. Sure, our lawyers put in a bit of reasonable legal fine print (you can’t resell the exact same product, you can’t use it for direct competition, you have to give us some acknowledgement, etc…); but other than that, we really do mean that these products are completely IP-Free. For example, you can use them in your research or create products to which you own all the rights.
See the complete legal IP-Free Terms and Conditions
DNA2.0 has patent applications filed for the Electra® cloning system using the type IIs enzyme SapI, and holds all rights for the system.
“Nothing like adding a little more tragedy to science!” – Dr. Aaron Straight, Stanford University

True, science can occasionally feel like a Greek tragedy. We named our new cloning system ‘Electra’ because it allows you to select against the pMOTHER vector just as Electra destroyed her own mother Clytemnestra. Hopefully, your endeavours are more successful than those of Antigone. Thankfully, unlike the original Electra, your selection of the daughter against the mother will not require any atonement to the gods.

Learn more about the classic Greek tragedy Electra:
Electra story – Euripedes version
Electra story – Sophocles version

Yes! See the protocol for all the necessary info, including primers and design. You can clone your PCR product into any pMOTHER or pDAUGHTER vector.

To PCR your ORF: we recommend you add the following ends to your primers, as these contain the Electra sites to clone directly into Electra vectors. Add 15-20 bp of your ORF to the 3′ primer end to amplify your ORF and have it compatible with any of the Electra MOTHER or Electra DAUGHTER expression vectors.

  • Forward primer:
    5′-TACACGTACTTAGTCGCTGAAGCTCTTCTATG….(ORF)….
  • Reverse primer:
    5′-AGGTACGAACTCGATTGACGGCTCTTCTACC….(ORF Reverse Complement)….

Use the DNA2.0 Bioinformatics Toolbox to facilitate primer analysis and design.

Your ORF must NOT contain any SapI recognition sites, as the Electra cloning process utilizes the type IIs enzyme SapI

Easy, simply PCR out your ORF using the PCR primer ends shown below and clone into any Electra vector using the standard protocol.

To PCR your ORF we recommend you add the following ends to your primers, as these contain the Electra sites to clone directly into Electra vectors. Add 15-20 bp of your ORF to the 3′ primer end to amplify your ORF and have it compatible with any of the Electra MOTHER or Electra DAUGHTER expression vectors.

  • Forward primer:
    5′-TACACGTACTTAGTCGCTGAAGCTCTTCTATG….(ORF)….
  • Reverse primer:
    5′-AGGTACGAACTCGATTGACGGCTCTTCTACC….(ORF Reverse Complement)….

Use the DNA2.0 Bioinformatics Toolbox to facilitate primer analysis and design.

Your ORF must NOT contain any SapI recognition sites, as the Electra cloning process utilizes the type IIs enzyme SapI

Yes! You can use the Electra reagents kit for all Electra cloning reactions.
Easy, DNA2.0 can modify almost any cloning or expression vector into an Electra Vector. Your favorite vector can then be used with the Electra system to quickly and efficiently transfer genes. Of course, your vectors remain your property, and the entire system is IP-Free.

Consult with an Electra Specialist today at +1 877 362 8646 or info@DNA20.com to learn more about improving your cloning and expression system.

Please note: the Electra Vector System is currently available to ship only to customers in the US, Canada, Japan, and major European hubs.
Electra System international shipping is available to European locations from our EU distributor, Cambridge Bioscience, Ltd.
Electra System international shipping is available to Japanese locations from our distributor, Cosmo Bio Co, Ltd.

Gene Synthesis Cloning orders in the Electra System® are available for all customers worldwide.

Yes!
The individual color names in the Protein Paintbox derive from our 2011 Holiday publication, Nine improved monomeric fluorescent proteins from Rangifer tarandus, where the IP-Free proteins were named after Santa’s reindeer (Rudolph, Donner, Comet, etc…). As the DNA2.0 fluorescent and chromogenic proteins became available for sale, we kept the reindeer names. As we added new colors to our product line, we continued with the holiday naming theme, branching out to include a diversity of cultural winter traditions, classic holiday literature, drama, and of course, television cartoons. We hope you enjoy this naming scheme.
Should you have any questions regarding the protein names or wish to suggest additional names, please email us at communications@dna20.com.
Multiple CFPs, GFPs, etc. have been selected to cover a range of intensities. For example, FrostyCFP – 20000, CindyLouCFP – 35000, TwinkleCFP – 12000.
No, the Paintbox proteins need oxygen to mature, so cannot be expressed anaerobically.
The Biobrick set (3 FPs) have been donated by DNA2.0 to the BioBricks Foundation and are covered under the Biobricks licensing agreement (BLA). These belong to BioBricks, and are therefore not included in the DNA2.0 ProteinPaintbox set.
The green/yellow FPs share only 27% homology at the amino acid level. Thus, it is fair to say that they are all unique proteins.
In the DNA2.0 labs, we get colored colonies in 24hrs.  As for how fast the maturation is, we have not done a time-course analysis. So, we cannot say if color develops in an hour or if it takes longer. Maturation time at 37°C – 24 hrs incubation is fast, 24-72 hrs is medium.
DNA2.0 offers an antibody (anti-CometGFP, catalog #AB-01) that will detect GFP which could be used to determine the number of GFP molecules. However, we do not currently have data for this. Note, since this is a polyclonal Ab, it should be able to detect any of our GFPs.
Maturation time and intensity varies for each of the proteins.
For example, the intensity of RudolphRFP is lower than DasherGFP, while maturation time for RudolphRFP is longer than for DasherGFP. Thus at an early timepoint, you may have the appearance of excellent DasherGFP expression, while only minimal RudolphRFP expression.
We have not measured half-life.
However, we have looked at color after a week and it appears to remain stable.
These proteins should be able to express in any genetic background that will support T5 driven expression.
Yes, they should mature in minimal media induced with IPTG.
Sure, you can learn more about vector design with T5 promoter, lac operator for IPTG induction and increased protein expression in bacterial systems here.
E.coli does not typically secrete proteins extracellularly, though some strategies can be employed. Please see below.

From: J. H. Choi . S. Y. Lee (2003). Metabolic and Biomolecular Engineering National Research, pp 630-632. “A number of methods have been used to promote extracellular secretion of recombinant proteins from E.coli. These include the use of biochemicals, physical methods (osmotic shock, freezing and thawing), lysozyme treatment and chloroform shock. E. coli normally does not secrete proteins extracellularly except for a few classes of proteins such as toxins and hemolysin. Secreted proteins can leak from the periplasmic space into the culture medium possibly due to an increased permeability of the cell membrane during a lengthy incubation period. Small proteins secreted into the periplasm are frequently released into the culture medium (Tong et al. 2000). In general, movement of recombinant proteins from the periplasm to the culture medium is the result of compromising the integrity of the outer membrane. However, care must be exercised during such recombinant protein production so as not to compromise cellular integrity, which often causes cell death. Interestingly, glycine or Triton X-100 supplemented to the medium retarded formation of inclusion bodies in the periplasm and increased the extracellular production efficiency of recombinant proteins (Jang et al. 1999; Kaderbhai et al. 1997; Yang et al. 1998). Glycine has been found to induce morphological changes, such as an enlarged spheroidal morphology in E. coli, as it is incorporated into peptidoglycan. Glycine supplementation may slightly disrupt peptidoglycan cross-linkages and cell membrane integrity. Yang et al. (1998) reported that adding 2% (w/v) glycine dramatically increased extracellular production of sFV/TNF-α and β-glucosidase.”

Yes, having the secretion signals can give enhaced expression in the cytoplasm even if the amount secreted or transported to the periplasm is limited.
At high rhamnose concentrations, we are seeing similar amounts of protein to T5. However, T5 might be somewhat stronger.
For standard applications we induce by adding rhamnose at mid-log just as we do with IPTG.
In E. coli, glucose shuts down expression very well, even in the presence of rhamnose. This is why it works for autoinduction. You add both and once glucose is consumed rhamnose induces.
No, our rhamnose vectors do not carry the rhaRS gene and it does not affect expression in wild type E.coli.