Difference between revisions of "Part:BBa M50056"

Revision as of 05:23, 12 December 2016

PETase double mutant I208V and R90A full plasmid information

We use Escherichia coli as the chassis organism to express the PETase gene and optimize our codons accordingly. This composite part is made of a rhamnose-inducible promoter, an ampicillin resistance marker, an origin of replication, a secretion tag, a strong RBS, and a PETase gene that contains two mutations. All basic parts except mutated PETase come from DNA 2.0. Wild type PETase shows the ability to hydrolyze Polyethylene terephthalate (PET), commonly used for plastics. The two mutations of PETase are identified by the 2016 iGEM Tianjin team to improve the catalytic activity of PET independently. The PETase gene here also incorporates 6 his tag at the end.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 633
Illegal PstI site found at 864
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 633
Illegal PstI site found at 864
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 633
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 633
Illegal PstI site found at 864
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 633
Illegal PstI site found at 864
Illegal NgoMIV site found at 220
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 388

@@ Line 3: / Line 3: @@
 <partinfo>BBa_M50056 short</partinfo>
-We use Escherichia coli as the chassis organism to express the PETase gene and optimize our codon accordingly. This composite part is made of a rhamnose-inducible promoter, an ampicillin resistance marker, an origin of replication, a secretion tag, a strong RBS, and a PETase gene that contains two mutations. All basic parts except mutated PETase come from DNA 2.0.
+We use Escherichia coli as the chassis organism to express the PETase gene and optimize our codons accordingly. This composite part is made of a rhamnose-inducible promoter, an ampicillin resistance marker, an origin of replication, a secretion tag, a strong RBS, and a PETase gene that contains two mutations. All basic parts except mutated PETase come from DNA 2.0.
-Wild type PETase shows the ability to hydrolyze Polyethylene terephthalate (PET), commonly used for plastics. The two mutations of PETase are identified by the 2016 iGEM Tianjin team to improve the catalytic activity of PET independently. The PETase gene here also incorporates 6 his tag at the end. The sequence is optimized for Escherichia coli.
+Wild type PETase shows the ability to hydrolyze Polyethylene terephthalate (PET), commonly used for plastics. The two mutations of PETase are identified by the 2016 iGEM Tianjin team to improve the catalytic activity of PET independently. The PETase gene here also incorporates 6 his tag at the end.
 <!-- Add more about the biology of this part here