Part:BBa_I713001

A pBBR1MCS plasmid without the mob gene.

This plasmid is constructed by PCR with the pBBR1MCS plasmid as template and the following primers:

DMOBF: AATTGCTAGCTGACGTACTCGACCGCCAACACAGC DMOBR: AATAGCTAGCATCCCAAGAGACGGCCCGAGACCTTC

The PCR product was phosphorad and ligated without enzyme digestion, although we did designed an NheI site in the primers.

Since the contruct process contains PCR amplification with Takara LA Taq. The product might have mutations.

The desire sequence should be:

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 25
  Illegal EcoRI site found at 772
  Illegal XbaI site found at 4172
  Illegal SpeI site found at 1
  Illegal PstI site found at 19
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 25
  Illegal EcoRI site found at 772
  Illegal NheI site found at 1484
  Illegal NheI site found at 1498
  Illegal SpeI site found at 1
  Illegal PstI site found at 19
  Illegal NotI site found at 4164
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 25
  Illegal EcoRI site found at 772
  Illegal BamHI site found at 7
  Illegal XhoI site found at 58
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 25
  Illegal EcoRI site found at 772
  Illegal XbaI site found at 4172
  Illegal SpeI site found at 1
  Illegal PstI site found at 19
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 25
  Illegal EcoRI site found at 772
  Illegal XbaI site found at 4172
  Illegal SpeI site found at 1
  Illegal PstI site found at 19
  Illegal NgoMIV site found at 3525
  Illegal AgeI site found at 3685
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1461