Part:BBa_J64100

Medium copy ColE1 BioBrick vector

All ccdb parts have been withdrawn from the Registry. Samples of parts containing the ccdB gene cannot be requested. - iGEM HQ

This plasmid is based on pProTetE333 (clontech). A SpeI site was removed via an A-T mutation at position 1166 on the original plasmid, resulting in pProTetE333b. The G00100-G00101 fragment of pSB4A3-1 which includes P1010 between a biobricks multiple cloning site which itself is flanked by + strand transcriptional terminators was then cloned between the XhoI and AvrII sites of pProTetE333b. The resulting plasmid carries a ColE1 origin of replication, a Chloramphenicol resistance marker, the biobricks MCS and the ccdAB death cassette.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal prefix found at 124
  Illegal suffix found at 821
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 124
  Illegal SpeI site found at 822
  Illegal PstI site found at 836
  Illegal NotI site found at 130
  Illegal NotI site found at 829
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 124
  Illegal BamHI site found at 809
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found at 124
  Illegal suffix found at 822
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found at 124
  Plasmid lacks a suffix.
  Illegal XbaI site found at 139
  Illegal SpeI site found at 822
  Illegal PstI site found at 836
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal BsaI.rc site found at 259