TMEM74 - ORF Clones

An ORF is a protein-coding sequence starting with an ATG start codon, ending with a stop codon, and devoid of both the 5’ and 3’ untranslated regions (UTRs).

An ORF clone is the ORF sequence pre-cloned in either a HaloTag® (FHC clones) or an untagged Flexi® (FXC clones) vector.

Nearly all HaloTag® ORF clones (FHC) are provided in the pFN21A vector, where HaloTag® is oriented at the N-terminus of the ORF sequence. The clone can easily be transferred to alternate Flexi® vectors, or you can request a custom clone to be created.

Nearly all Native ORF clones (FXC) will be provided in the pF1K vector. Again, the clone can easily be transferred to alternate Flexi® vectors, or you can request a custom clone to be created.

The majority of ORF clones reach our customers within three weeks. However, please allow up to 4-6 weeks for delivery.

You will receive 100ng of purified plasmid DNA in T.E. It is highly recommended that you transform your ORF clone into competent cells, and create a bacterial glycerol stock or additional purified DNA prior to exhausting the 100ng received. If you would like a greater quantity of DNA, please request a custom clone.

All ORF clones undergo quality testing to confirm size and sequence identity. In addition, nearly all HaloTag® ORF clones are tested for protein expression and fluorescent microscopy images in HEK293 cells when produced. This data is available in the Figures/Sequence tab.

The ORF Clone will be shipped to your lab at ambient temperature. Once received, store the ORF Clone at -20°C for long-term storage, or at 2 – 8°C for immediate use.

Purified DNA in T.E. has been shown to be very stable over time. An expiration date of two years is put on all ORF Clones when produced, but the clone may still be viable years afterward. Once received, best practice suggests transforming your ORF clone into competent cells to create a safety glycerol bacterial stock(s), and/or additional aliquots of purified DNA prior to exhausting the 100ng received.

The HaloTag® ORF Clones (FHC) can be used in mammalian cells to express a HaloTag fusion protein from a CMV promoter. In addition, these clones afford expression in Cell-Free Expression systems (specifically: L1170, L1171, L4610, and L4611). However, the FHC ORF clones are not designed to express in E. coli, yeast, insect, or other expression systems. The ORF can easily be transferred to alternate Flexi® vectors as your expression system needs may change.

The Native ORF Clones (FXC) can be used in E. coli to express an untagged protein from a T7 promoter. In addition, these clones afford expression in Cell-Free Expression systems (specifically: L1170, L1171, L4610, L4611, L1110, L1115, and L1130). However, the FXC ORF clones are not designed to express in mammalian cells, yeast, insect, or other expression systems. However, the ORF can easily be transferred to alternate Flexi® vectors as your expression system needs may change.

Many customers have specific requests regarding an ORF clone. Common requests include: generating a new ORF clone to a human or mouse gene, transferring an existing ORF clone into a different Flexi® vector, requesting a larger quantity of DNA, or even requesting a custom cell line from an ORF clone. All these requests and more can be accommodated. Please let us know what you need by completing a custom clone request form. Pricing and delivery times will vary based upon request.

Contact us at proteomics@promega.com, or speak to your local Promega representative. Discounts will be provided based upon scale and format needs.

If you encounter issues with the integrity of the clone received, please contact Kazusa DNA Research Institute at office@kazusa.or.jp for troubleshooting ORF clone questions. If you are unsatisfied with your response, please contact Promega at techserv@promega.com. All questions about the HaloTag® technology should be directed to Promega.

KDRI is an academic Japanese research institute dedicated to genome-scale studies and the function of individual genes. The Kazusa ORFeome Project was initiated to create a collection of human and mouse ORFs to enable the larger research community with a readily accessible, highly validated ORFeome. KDRI is an active member of the global ORFeome Collaboration, and are experts in HUGE (Human Unidentified Gene-Encoded) large proteins, >4 kb. Promega and KDRI have established a partnership, allowing Promega customers access to Kazusa's coveted collection of experimentally validated clones.

HaloTag® and Native ORF clones are strictly to be used for research purposes only. Persons wishing to use this product or its derivatives in other fields of use, including without limitation, commercial sale, diagnostics or therapeutics, should contact Promega Corporation for licensing information. Please refer to the HaloTag® Limited Use Label Licenses, and other documentation provided at shipment with the clone for further details.