Part:pSB4C1

pBR322BB2

Plasmid pCK01BB1 (Fig. 1) is based on pCK01 (Fig. 2), a low-copy cloning vector (5-12 copies per cell) that confers chloramphenicol resistance (CmR) and harbours the pSC101 origin of replication.

Fig. 1: Map of the pCK01BB1 plasmid

Fig. 2: Map of the original pCK01 plasmid

In order to make this plasmid compatible with the BioBrick Standard assembly process, the original pCK01 was digested at its AgeI and AseI restriction sites and an appropriate multiple cloning site (MCS) was introduced in this position. Since the original plasmid also contains a SpeI restriction site in the origin of replication, the ACT codon at this position was changed to ATT by site directed mutagenesis. The term "BB1" in pCK01BB1 refers to "BioBrick version 1".

Purpose

This plasmid was designed for the [http://parts.mit.edu/igem07/index.php/ETHZ ETHZ iGEM 2007 project] and is used for the following BioBricks: Part 4, Part 5new, Part 8, Part 9 and the composites BBa_I739015, BBa_I739016 and BBa_I739017.More information on the plasmid and the assembly strategy of the used parts can be found [http://parts.mit.edu/igem07/index.php?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