You successfully amplify the psy2 sequence and wish to clone

You successfully amplify the psy2 sequence and wish to clone the PCR fragment into vector 2 in order to express it in yeast. The cloning sites available on this vector are shown below.

There are two different ways to insert the amplified psy2 sequence into vector 2. Give the
restriction enzyme(s) that you could use to cut the vector and the psy2 coding sequence for each
of these.

3. Directional cloning:
Cut vector and psy2 with:
4. Non directional cloning:
Cut vector and psy2 with:
5. Which enzyme (s) could you use to verify the presence and orientation of the insert?

Stu Promoter Sall EcoR Start EcoRI 5 GTC GAC G GAA TTC AGGCCT C GTC GACT CCGGC3 Psy2 Vector 2 3\' CAGCT GCC TTA AGT CCG GA L G CAG CT GAGGCCG5 StuI: 5\'-AGG/CCT-3 SalI: 5\'-G/TCGAC-3 EcoRI: 5\'-G/A ATTC-3\' BamHI: G/GA TCC

Solution

2)

Answer:

The restriction enzymes to cut the vector 2 would be StuI , SalI and EcoRI

The restriction enzymes to cut the psy2 would be StuI , SalI

3)

Answer:

Directional cloning:
Cut vector and psy2 with: StuI and SalI

4)

Answer:

Non directional cloning:
Cut vector and psy2 with: SalI and StuI

5)

Answer:

The enzyme could be used to verify the presence and orientation of the insert is EcoRI