Part:pSB3K1

pACYC177 converted into BioBrick vector (formerly BBa_I739202)

Plasmid pACYC177BB1 (Fig. 1) is based on pACYC177 (Fig. 2), a low-copy cloning vector (10-12 copies per cell) that confers ampicillin (ApR) and kanamycin (KmR) resistance and harbours the p15A origin of replication.

Fig. 1: Map of the pACYC177BB1 plasmid

Fig. 2: Map of the original pACYC177 plasmid

In order to make this plasmid compatible with the BioBrick Standard assembly process, the original pACYC177 was digested at its BamHI 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 kanamycin resistance is left. The term "BB1" in pACYC177BB1 refers to "BioBrick version 1".

Purpose

This plasmid was designed for the [http://2007.igem.org/ETHZ ETHZ iGEM 2007 project] and is used for the following BioBricks: Part 6, Part 7, Part 10, Part 11 and the composites BBa_I739018, BBa_I739019 and BBa_I739020. 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
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 3043
  Illegal NheI site found at 1305
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NotI site found at 9
  Illegal NotI site found at 3049
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 3043
  Illegal BamHI site found at 3037
  Illegal XhoI site found at 1669
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found at 3043
  Illegal suffix found at 2
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found at 3043
  Plasmid lacks a suffix.
  Illegal XbaI site found at 3058
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal AgeI site found at 895
  Illegal AgeI site found at 1219
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal BsaI site found at 190