Part:BBa_K3963001

pBAV1k-lacI-Trc-beta-agarase YM01-3

The plasmid is combined by the backbone from the pBWB162 plasmid (BBa_K3963003) and cloned by restriction based cloning with the insert beta-agarase YM01-3 (BBa_K2094002). mCherry protein was cut out. The plasmid still contains the lacI behind lacIq promoter and a kanamycin selection marker.

Design

Figure 1 Plasmid pBAV1k-lacI-Trc-beta-agarase YM01-3 design. The plasmid size is 5560 bp.

Experiments and results

Plasmid

The plasmid should have a size of 5560 bp but in the gel the plasmid looks smaller about 4 kb (see Fig. 2A).We transformed the plasmid pBAV1k-lacI-Trc-beta-agarase YM01-3 into E. coli DH5ɑ and the typical pit formation can be observed (see Fig. 2B). We send the plasmid to seuquencing to control the beta-agarase insert (see Fig. 2B).

Figure 2

Natural transformation function

In Figure 3 it can be seen that after cloning of the pBWB162 plasmid (BBa_K3963000) to the recent plasmid the ability to be taken up by Acinetobacter baylyi ADP1, a natural competent bacterial strain, is still existing.

Figure 3

Discussion

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Unknown
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 3183
  Illegal AgeI site found at 3220
  
• 1000
  COMPATIBLE WITH RFC[1000]