You are a helpful assistant. When the user asks a question, your response must include two parts: first, the reasoning process enclosed in <think>...</think> tags, then the final answer enclosed in <answer>...</answer> tags.Please provide a clear, concise response within <answer> </answer> tags that directly addresses the question.
system
<image>Consider the following plasmid, pET28a, a popular expression vector that is used for protein expression in bacteria: <image 1>. You freshly purified plasmid from overnight bacterial culture using extraction kit, also known as miniprep. You confirmed the quality of plasmids by checking DNA and protein ratio (A260/280) and running them on a DNA agarose gel. They looked pure as they should be. You would now like to digest this vector for gene manipulation (molecular cloning) to generate vectors suitable for molecular cloning. If you cut the above plasmid with restriction enzyme called HindIII, the form of DNA will be changed from _________
A. Linear to supercoiled
B. Nicked to linear
C. Nicked to supercoiled
D. Supercoiled to nicked
E. Supercoiled to linear

Please answer the question with option letter from given choices in the format of Answer: <option letter>. You FIRST think about the reasoning process as an internal monologue and then provide the final answer. The reasoning process MUST BE enclosed within <think> </think> tags. Please answer the final answer within the <answer> </answer> tags.
user
E
Consider the following plasmid, pET28a, a popular expression vector that is used for protein expression in bacteria: <image 1>. You freshly purified plasmid from overnight bacterial culture using extraction kit, also known as miniprep. You confirmed the quality of plasmids by checking DNA and protein ratio (A260/280) and running them on a DNA agarose gel. They looked pure as they should be. You would now like to digest this vector for gene manipulation (molecular cloning) to generate vectors suitable for molecular cloning. If you cut the above plasmid with restriction enzyme called HindIII, the form of DNA will be changed from _________
A. Linear to supercoiled
B. Nicked to linear
C. Nicked to supercoiled
D. Supercoiled to nicked
E. Supercoiled to linear

Please answer the question with option letter from given choices in the format of Answer: <option letter>. You FIRST think about the reasoning process as an internal monologue and then provide the final answer. The reasoning process MUST BE enclosed within <think> </think> tags. Please answer the final answer within the <answer> </answer> tags.
val