Part:BBa_K200009

DpnII Composite

This sequence codes for the restriction enzyme DpnII.

Detail

This is a Type II restriction enzyme that recognises the nucleotide sequence 'GATC.' Its activity is blocked by dam methylation.

The working temperature is 37°C.

The gene was used alongside with the restriction enzyme TaqI and the methylase Dam by the Imperial iGEM 2009 [http://2009.igem.org/Team:Imperial_College_London The E.ncapsulator] project as part of the genome deletion module.

DpnII is a 4 base cutter, leaving a 4 base overhang.

The cutting site is as shown:

Usage and Biology

Enzyme Properties can be viewed at the [http://www.neb.com/nebecomm/products/productR0543.asp DpnII Datasheet] from New England Biolabs

Methylation Sensitivity: dam methylation: Blocked dcm methylation: Not sensitive CpG methylation: Not sensitive More information about: Methylation Sensitivity

Heat Inactivation: 65°C for 20 minutes

Survival in a Reaction: (+ + +) Suitable for an extended or overnight digestion. Enzyme is active > 8 hours. More information about: Extended Digests with Restriction Enzymes

Sequence and Features

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

References

[http://www.neb.com/nebecomm/products/productR0543.asp Information from NEB]