Custom Gene Synthesis (GS0020)

May Promotion:

 

Price list:
GeneSynthesis_prices

Description: NZYTech’s Custom Gene Synthesis service provides in vitro chemical synthesis, cloning and 100% sequence verification of any desired DNA genetic sequence. NZYTech’s R&D department improved standard molecular biology processes and developed innovative bio-informatics tools for gene design and codon optimization. Our proprietary gene optimization algorithm – ATGenium – comprehensively optimizes critical factors in transcription, translation and co-translational protein folding to deliver highest levels of expression in any host system. Genes are cloned into our standard cloning vectors. Downstream services, including sub-cloning, site directed mutagenesis, plasmid prep and protein expression are available upon request. Sub-cloning into one of our pHTP Escherichia coli expression plasmids – pHTP expression vectors – is provided at a very competitive price.

Features:
– Flexible, custom gene design process: DNA sequences can be engineered to add tags, restriction sites, promoters or other elements
– Free gene design & codon optimization – ATGenium – can be used to optimize DNA sequences and enhance protein expression, if requested
– All genes are 100% sequence verified and cloned into our standard cloning vectors – pHTP0 or other
– Possibility to sub-clone your gene into any destination vector
– Complete confidentiality guaranteed
– Quality assurance certificate
– Competitive prices

Complementary services (*):
– Your synthetic gene can be easily transferred from the standard entry vectors into one of our pHTP Escherichia coli expression plasmids for a very competitive price (see gene cloning). High protein expression levels can be reached when using pHTP expression vectors (see expression levels).
– Alternatively, sub-cloning your gene into your vector of choice is also possible (see gene cloning).
– Screening the best expression conditions for your recombinant protein is also available and may include gene sub-cloning into a battery of pHTP expression vectors (see figure below), followed by protein expression and purification (see enhanced protein expression).

 (*) at additional price

pHTP0_pHTPExpression_Transfer_Nov2017

Synthetic genes cloned into pHTP0 cloning vector can be easily transferred into any pHTP E. coli expression vector. The portfolio of pHTP expression vectors includes a different range of fusion tags, thus offering the possibility to quickly assay levels of expression and solubility of a desired protein in multiple expression vectors.

Delivery Specifications:
– 0.5-1 μg purified recombinant plasmid containing your synthesized gene
– Gene Synthesis Report containing vector map, gene sequence and QC certificate
– Delivery in 30 to 35 working days for non-complex genes

Quotes & Ordering: 
For quotations and orders, please email us to services@nzytech.com; send us your gene sequence in FASTA format, or upload and fill out the NZYTech’s Gene Synthesis Order Form. A quote, together with the corresponding gene synthesis project, will be send to you within 24h. Our dedicated PhD-level project management team can tailor the order according to your specific requirements, and support you throughout the entire project.

Prices and turnaround times are estimated for non-complex genes and may increase as gene length and complexity increases. Sequences that are unstable and/or toxic to E. coli can substantially affect delivery times.

Please contact us to learn about discounted pricing for high-volume orders, institutional contracts, long-term partnerships, or to discuss your specific requirements.

Additional information:
– Your gene request will be reviewed by our experienced scientists in order to investigate the presence of elements that may interfere with synthesis and/or assembly performance. Design warnings include:

– Secondary structure: hairpins; local and global GC content
– Repeat content; homopolymers: A/C/T <=10 bases and G <=6 bases
– Restriction sites

– You will receive a gene synthesis project describing your detailed request. If gene sequence does not pass the screening criteria, our gene services specialists will contact you to determine the best way to proceed.
– Downstream services also available upon request (gene sub-cloning; site directed mutagenesis & recombinant protein production).
– Beware that higher DNA yields and endotoxin free DNA may be provided upon request at an additional cost.
– Please notice that each gene will be screened to identify potentially dangerous pathogen sequences. NZYTech reserves the right to refuse any order that does not pass this biosafety screen.

pHTP0 vector map
pUC57 vector map
Order Form
Codon optimization of your genes

Proteins play a variety of critical roles in many biological processes. They are valuable therapeutic compounds, research reagents and industrial catalysts. NZYTech’s proprietary codon optimization software – ATGenium – can alter both naturally occurring and recombinant gene sequences to achieve the highest possible levels of protein expression in any given expression system. This will save you time and money, while getting you quickly to meaningful results. ATGenium takes into consideration a variety of critical factors involved in different stages of protein expression, such as codon adaptability, mRNA structure, and various cis-elements in transcription and translation. Optimizing your gene sequence will have significant advantages in a variety of applications such as recombinant protein expression, DNA vaccine design and protein engineering.

Optimizing heterologous protein production in Escherichia coli with ATGenium:
Superior levels of recombinant proteins expressed in Escherichia coli strains can be achieved through codon optimization using the NZYTech’s ATGenium algorithm.

ATGenium_expression_levels

A. Levels of protein expression of three different eukaryotic proteins in a prokaryotic system (Escherichia coli). The genes encoding these proteins were optimized by three algorithms for codons optimization, by two different competitors (light blue) and using the ATGenium algorithm developed by NZYTech (dark blue).

B. High-throughput recombinant protein expression of CAZYmes (Carbohydrate Active eNZymes). Genes encoding these proteins were optimized using the ATGenium algorithm, cloned into pHTP1 expression vector and overexpressed incoli BL21 (DE3) strain. Recombinant proteins were purified through IMAC and separated by SDS-PAGE. High protein expression yields were obtained when genes were optimized using the ATGenium algorithm

1. Why should I use NZYTech’s gene synthesis services?
The key advantages of producing genes synthetically are the following:

– Less expensive than traditional cloning.
– You can spend your time on other projects while your gene is being produced.
– It is possible to codon optimize your gene, thus increasing the possibility of achieving high protein levels.
– Most DNA sequences can be synthesized.
– Gene synthesis provides access to DNA templates that are not easily available on nature.

2. How long does it take to synthesize a gene?
It usually takes 20 to 35 working days to synthesize most genes. However, sequences that are unstable and/or toxic to E. coli can substantially affect delivery times. Delivery times may increase as gene length and complexity increases.

3. Is it possible to synthesize sequences with high GC content or repeats?
Sequences with high and low GC content are complex to be handled and usually require a longer production period. However, NZYTech has successfully synthesized genes with high/low GC contents. Please contact us to carefully review every sequence; we will provide you a custom gene synthesis project, which includes turnaround time and sequence details.

4. Is it possible to optimize gene design for high levels of expression?
ATGenium algorithm optimizes different factors that are known to affect mRNA stability, such as GC content and secondary structures. It allows optimizing not only critical factors in transcription and translation but also factors related with co-translational protein folding. The relative frequency of codon usage varies widely depending on the organism. ATGenium comprises an extensive collection of codon usage tables, allowing optimizing codon usage for improved protein expression in different host systems.

5. In which vector am I going to receive my gene?
If you have not requested sub-cloning of your gene into a specific vector, you will receive your gene cloned into our standard cloning vectors (pHTP0 or other). Our cloning vectors are ampicillin resistant. Vector maps are available on the website. We can easily transfer your gene into one or more pHTP expression vector(s) for a very convenient price.

6. Are all genes sequence-verified?
All genes are 100% sequence verified.

7. Is it possible to sub-clone my gene into a commonly used expression vector?
Your synthetic gene can be easily transferred from the entry vector to any of our pHTP E. coli expression vectors for a very competitive price. Alternatively, we can sub-clone your gene into your vector of choice.

8. How do you deliver synthetic genes?
NZYTech delivers 0.5 to 1 µg of plasmid DNA.

9. What is NZYTech’s policy for Intellectual Property Protection?
NZYTech does not claim rights or ownership of any intellectual property related to the DNA/amino acid sequences provided by our customers or the resulting synthetic genes. No data or material will be released to a third party without your consent. All genes resulting from gene synthesis and/or cloning services are stored at NZYTech for a three months period. After this period nucleic acids provided to customers are discarded.

Get a Free Discount Coupon if your paper refers NZYTech Products!

If NZYTech’s products are mentioned in a paper published by your lab, you will be rewarded with a Free Discount Coupon of 50 € to use in any order. Please note that the final paper/page proof (as PDF files) or a downloadable link must be provided so NZYTech can confirm that your submission is valid for this promotion.

See more details here.

 

Featured citations

The gene for human Galectin-3 was synthesized by NZYTech using a codon optimization strategy for E. coli expression. The region corresponding to residues 114-250 (named Gal-3 CRD) was sub cloned into the expression vector pET21 by NZYTech.
Diniz, A., Dias. J.S., Jiménez-Barbero, J., Marcelo, F., Cabrita, E. J. (2017). Protein-Glycan Quinary Interactions in Crowding Environment Unveiled by NMR Spectroscopy. Chemistry 23(53):13213-13220

 

Alternatively, gene butA fused with 185 bp region upstream of tuf gene (Ptuf) was synthesized by NZYTech (Lisbon, Portugal).
Rados, D. et al (2015). Engineering Corynebacterium glutamicum for the production of 2,3‑butanediol. Microbial Cell Factories 14:171

 

TAL, C3H, DCS, CURS1, CUS, and CCoAOMT genes were codon optimized for E. coli, synthesized and cloned in the plasmid vector pUC57 by GenScript or NZYTech (Lisbon, Portugal).
Rodrigues, J. L., Araújo, R. G., Kristala, L. J. P., Leon, D. K., Rodrigues, L. R. (2015). Production of curcuminoids from tyrosine by a metabolically engineered Escherichia coli using caffeic acid as an intermediate. Biotechnol. J. 10:1-11

 

 

 

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