Difference between revisions of "Part:BBa K3963001"
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<partinfo>BBa_K3963001 short</partinfo> | <partinfo>BBa_K3963001 short</partinfo> | ||
− | The plasmid is combined by the Backbone from the pBWB162 plasmid (BBa_K3963000) and cloned by restriction based cloning with the insert beta- | + | The plasmid is combined by the Backbone from the pBWB162 plasmid (BBa_K3963000) 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== | |
− | [[File:T--Heidelberg--beta-agarase--NT--Map.png|thumbnail|center|500px|<b>Figure 1 Plasmid design</b>]] | + | [[File:T--Heidelberg--beta-agarase--NT--Map.png|thumbnail|center|500px|<b>Figure 1 Plasmid pBAV1k-lacI-Trc-beta-agarase YM01-3 design.</b> 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 <I>E. coli</I> 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== | ||
+ | |||
+ | Figure 3 | ||
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Revision as of 10:19, 19 October 2021
pBAV1k-lacI-Trc-beta-agarase YM01-3
The plasmid is combined by the Backbone from the pBWB162 plasmid (BBa_K3963000) 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
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
Figure 3
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2292
Illegal BamHI site found at 3167 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 3183
Illegal AgeI site found at 3220 - 1000COMPATIBLE WITH RFC[1000]