Biochemistry 4th Edition by Reginald Garrett, Charles Grisham

Exercise 3

Design a vector polylinker for directional cloning
A vector has a polylinker containing restriction sites in the following order: Hin dIII, Sac I, Xho I, Bgl II, Xba I, and Cla I.
a. Give a possible nucleotide sequence for the polylinker.
b. The vector is digested with Hin dIII and Cla I. A DNA segment contains a Hin dIII restriction site fragment 650 bases upstream from a Cla I site. This DNA fragment is digested with Hin dIII and Cla I, and the resulting Hin dIII- Cla I fragment is directionally cloned into the Hin dIII- Cla I digested vector. Give the nucleotide sequence at each end of the vector and the insert and show that the insert can be cloned into the vector in only one orientation.