Part:BBa_K2151666
pMG36ET
This is a plasmid backbone consisting of a modified version of the S. thermophilus expression vector pMG36ET. The multiple cloning site was replaced with the biobrick prefix and suffix. The backbone contains the pWV01 broad-host range origin of replication, making it a suitable shuttle vector for use within E. coli and lactic acid bacteria such as S. thermophilus. The backbone also contains an erythromycin resistance gene as selectable marker.
<p<We were able to transform both E. coli and S. thermophilus with this plasmid, resulting in erythromycin-resistant colonies. By using this plasmid as a shuttle vector, we were able to clone BioBrick constructs in E. coli, and then purify the DNA for transformation of S. thermophilus. We successfully transformed S. thermophilus with an amilCP BioBrick construct in this shuttle vector,confirming that the plasmid is suitable for use in both E. coli and S. thermophilus</p>
The part contains BBa_J04450 as is required by iGEM for submission of plasmid backbones.
Figure 1: Left: empty pMG36ET with BioBrick restriction sites highlighted Right: pMG36ET with J04450 BioBrick construct inserted between existing EcoRI and PstI restriction sites.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 3639
Illegal NheI site found at 2426
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 3645 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 3639 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found at 3639
Illegal suffix found at 2 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found at 3639
Plasmid lacks a suffix.
Illegal XbaI site found at 3654
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal AgeI site found at 447 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
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