Description

Human Open Reading Frame (ORF) Library contains approximately 15,000 individual gene ORF clones, including around 12,000 known genes. Each plasmid in this library is derived from a single clone, and every ORF has been sequence-verified. Subsequent cloning of the target gene into a new expression vector can be easily achieved using the Gateway system or PCR.

The clones in the ORF library are contained in the donor vector (pDONR223), as illustrated in Figure 1.

Figure 1: pDONR223 Plasmid Map. The full sequence information for pDONR223 can be found in the file: pDONR223-seq.txt

ORF Clones: These refer to the coding sequences between the start and stop codons, excluding the untranslated regions (UTRs) at the 5' and 3' ends of mRNA. The ORF library was created by reverse transcribing mRNA to obtain the cDNA encoding the protein and then cloning this into the pDONR223 vector using Gateway BP recombination technology, resulting in the entry vector. The recombination site sequence is shown in Figure 2. To facilitate the addition of appropriate 3' tags using the Gateway system, all clones lack stop codons. In the Beyotime Human Gene ORF library, each clone represents a unique insert sequence. Random clone sequencing accuracy is 95%, though the accuracy is slightly lower for longer gene clones.

Figure 2: Recombination Sequences Flanking the ORF

Our Human Gene ORF library provides E. coli glycerol stocks (DH5α) containing the respective ORF plasmids. This product is available in a set of 169 x 96-well plates. The cloning vector is pDONR223, which has spectinomycin resistance, with a recommended final concentration of 50 μg/ml. The forward sequencing primer (M13F) sequence is GTAAAACGACGGCCAGT, and the reverse sequencing primer (T7) sequence is TAATACGACTCACTATAGG.

Storage

• Store at -20℃ for up to two years.
• Store at -80℃ for up to five years.

Precautions

• The pDONR223 vector cannot directly express the corresponding protein in bacteria or mammalian cells; it must be recloned into the appropriate expression vector for protein expression.
• This product is intended for scientific research use by professionals only and is not to be used for clinical diagnosis or treatment, food or drug use, or stored in a residential environment.
• For your safety and health, please wear a lab coat and disposable gloves when handling.

Related: Gene Synthesis Services

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