pEBNA DEST Vector Kit

pEBNA DEST Vector Kit - FAQs

View additional product information for pEBNA DEST Vector Kit - FAQs (A10898)

4 product FAQs found

Where is the hygromycin gene from in pCEP4, pMEP4, pREP4, pREP7, pREP10, pEBVHis, and pRBK?

The hygromycin resistance gene in these vectors is driven by the TK (Thymidine Kinase) promoter and comes from E. coli.

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What is HAT?

HAT is a compete medium supplemented with 100 micromolar hypoxanthine, 0.4 micromolar aminopterin, 16 micromolar thymidine, and 3 micromolar glycine (HAT medium) is used when using selection vectors that encode for thymidine kinase (TK). For more information on this selection method, you can refer to Current Protocols in Molecular Biology ("Red Book") or the following reference:Littlefield, JW 1964 "Selection of hybrids from matings of fibroblasts in vitro and their presumed recombinants." Science 145:709-710.

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What is the promoter that drives the EBNA-1 gene?

The EBNA-1 gene in pCEP4 is expressed from a natural EBV promoter and TATA box. The promoter is not well defined, but if you BLAST the sequences upstream of the EBNA-1 ORF you will see that they line up with EBV genomic sequences.

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Where is the termination/polyA sequence for the EBNA-1 gene?

There is probably no termination/polyA for EBNA-1. There are only about 75 bases after the stop codon before the pUC sequence resumes with no obvious signal sequences.

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