Bio-Rad Ligation and Transformation Module User Manual

Page 22

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•

Its MCS has restriction enzyme sites that can be used for later
manipulation of the DNA

•

It is a high copy number plasmid

•

It contains the

b-lactamase gene, amp

r

, which confers resistance to

ampicillin

•

It contains the

eco47IR gene, which allows positive selection of

transformants. This gene codes for the

Eco47I restriction enzyme, and

when the enzyme is expressed, it is toxic to

E. coli. When the eco47IR

gene is disrupted by the insertion of DNA into the cloning site, the gene
will no longer be expressed and the transformed cells will grow and
divide on selective media

•

It is 2,974 bp long and its sequence is readily available — the accession
number for pJet1.2 is EF694056

pJet1.2 blunted vector.

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