What are the facts that determine plasmid copy number? #Everyone is a writer

Q: What are the facts that determine plasmid copy number?

    The copy number refers to the number of molecules of an individual plasmid that are normally found in a single bacterial cell. The factors that control copy number are not well understood. Some plasmids, especially the larger ones, are stringent and have a low copy number of perhaps just one or two per cell; others, called relaxed plasmids, are present in multiple copies of 50 or more per cell. Generally speaking, a useful cloning vector needs to be present in the cell in multiple copies so that large quantities of the recombinant DNA molecule can be obtained.

Plasmids vary widely in copy number depending on three main factors:

• 1) The ori and its constituents – (e.g. ColE1 RNA I and RNA II). to maintain the copy number,plasmids use definitive regulatory circuits.Negative regulators(inhibitors),acting at the initiation step,were involved in this control.
• 2) The size of the plasmid and its associated insert (bigger inserts and plasmids may be replicated at a lower number as they represent a great metabolic burden for the cell).
• 3) Culture conditions (i.e. factors that influence the metabolic burden on the host).