Part:BBa_I739203

pBR322BB2

Plasmid pBR322BB2 (Fig. 1) is based on pBR322 (Fig. 2), a low-copy cloning vector (15-20 copies per cell) that confers ampicillin (ApR) and tetracylin (TcR) resistance and harbours the pMB1 origin of replication.

Fig. 1: Map of the pBR322BB2 plasmid

Fig. 2: Map of the original pBR322 plasmid

In order to make this plasmid compatible with the BioBrick Standard assembly process, the original pBR322 was digested at its EcoRI and PstI restriction sites and an appropriate multiple cloning site (MCS) was introduced in this position. However, this leads to a loss of the original ampicillin resistance and only tetracyclin resistance is left. The term "BB2" in pBR322BB2 refers to "BioBrick version 2".

Purpose

This plasmid was designed for the [http://2007.igem.org/ETHZ ETHZ iGEM 2007 project] and was used as a backup plasmid for pBR322BB1. More information on the plasmid and the used assembly strategy can be found [http://2007.igem.org?title=ETHZ/Biology/Lab here].

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 3638
  Illegal SpeI site found at 3621
  Illegal PstI site found at 3607
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 229
  Illegal SpeI site found at 3621
  Illegal PstI site found at 3607
  Illegal NotI site found at 3612
  Illegal NotI site found at 3645
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 375
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 3638
  Illegal SpeI site found at 3621
  Illegal PstI site found at 3607
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 3638
  Illegal SpeI site found at 3621
  Illegal PstI site found at 3607
  Illegal NgoMIV site found at 401
  Illegal NgoMIV site found at 769
  Illegal NgoMIV site found at 929
  Illegal NgoMIV site found at 1283
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 3433
  Illegal SapI site found at 2350