Part:pSB4C1

pCK01 converted into BioBrick vector (formerly BBa_I739202)

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://2007.igem.org/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 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 3134
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 3140
21
INCOMPATIBLE WITH RFC[21]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 3134
23
INCOMPATIBLE WITH RFC[23]
Illegal prefix found at 3134
Illegal suffix found at 2
25
INCOMPATIBLE WITH RFC[25]
Illegal prefix found at 3134
Plasmid lacks a suffix.
Illegal XbaI site found at 3149
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal AgeI site found at 3128
1000
INCOMPATIBLE WITH RFC[1000]
Plasmid lacks a prefix.
Plasmid lacks a suffix.