DNA cloning and sequencing


You want to subclone the his operon from an F' plasmid to the small multicopy plasmid vector shown below. Given DNA from the F' plasmid and DNA from the vector plasmid, how could you make the recombinant plasmid and how would you select for cells that carry your subclone?


Certain restriction enzymes recognize different DNA sequences but generate compatible sticky ends (that is, the single stranded ends from DNA cut with the two different restriction enzymes can hybridize with each other). Which of the following restriction enzymes create compatible sticky ends and what is the DNA sequence of the compatible sticky end?

BamHIGGATCC
CCTAGG
EcoRIGAATTC
CTTAAG
BglIIAGATCT
TCTAGA


What would the following sequence look like on a sequencing gel using the Dideoxy technique? Be sure to label the lanes and indicate the direction of electrophoresis?

5' GGTACCTTAGC


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Last modified February 15, 1999