Part:pSB1AK3-1

PSB1AK3 with ccdB insert

Target plasmid for three antibiotic assembly. The ccdB insert prevents growth of the unmodified plasmid in normal cloning strains, allowing selection of only correctly assembled recombinants.

Usage and Biology

pSB1AK3 plasmid with a ccdB insert and constitutive promoter. The expressed ccdB protein is lethal to normal E. coli cells. This plasmid can only be grown in resistant cell strains, such as DB3.1

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal suffix found in sequence at 816
  Illegal EcoRI site found at 119
  Illegal XbaI site found at 134
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 119
  Illegal SpeI site found at 817
  Illegal PstI site found at 831
  Illegal NotI site found at 125
  Illegal NotI site found at 824
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 119
  Illegal BamHI site found at 804
  Illegal XhoI site found at 1850
  Illegal XhoI site found at 2876
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal suffix found in sequence at 817
  Illegal EcoRI site found at 119
  Illegal XbaI site found at 134
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 119
  Illegal XbaI site found at 134
  Illegal SpeI site found at 817
  Illegal PstI site found at 831
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 254
  Illegal BsaI.rc site found at 3022
  

Functional Parameters

chassis	DB3.1
copies	100-300
insert	ccdB
mcs	Biobrick
origin	pMB1
resistance	AK