Difference between revisions of "Part:BBa K4231019"
Line 4: | Line 4: | ||
Using pUC19 as plasmid backbone, AmpR as reporter gene in bacteria, LEU tag as reporter gene in Yarrowia lipolytica, shuttle plasmid carrying pTef1-BbTps3, pGAP-ERG20, pTef1-ERG12 from Yarrowia lipolytica. The plasmid can be effectively amplified in E.coli DH5a and can be successfully expressed in Yarrowia lipolytica. At the same time, ERG12 and ERG20 are important enzyme genes in the MVA pathway, and they carry the PTS1 sequence to help the enzyme protein locate in the peroxisome. Therefore, the plasmid plays an important role in the construction of MVA pathway in the peroxisome of Yarrowia lipolytica. | Using pUC19 as plasmid backbone, AmpR as reporter gene in bacteria, LEU tag as reporter gene in Yarrowia lipolytica, shuttle plasmid carrying pTef1-BbTps3, pGAP-ERG20, pTef1-ERG12 from Yarrowia lipolytica. The plasmid can be effectively amplified in E.coli DH5a and can be successfully expressed in Yarrowia lipolytica. At the same time, ERG12 and ERG20 are important enzyme genes in the MVA pathway, and they carry the PTS1 sequence to help the enzyme protein locate in the peroxisome. Therefore, the plasmid plays an important role in the construction of MVA pathway in the peroxisome of Yarrowia lipolytica. | ||
+ | [[file:PUC-BbTps3-ERG12-ERG20-LEU.png | border | center | 400px]] | ||
+ | |||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Revision as of 14:39, 7 October 2022
pUC19 carrying with ERG12, ERG20, BbTps3 and LEU tag from Y.lipolytica
Using pUC19 as plasmid backbone, AmpR as reporter gene in bacteria, LEU tag as reporter gene in Yarrowia lipolytica, shuttle plasmid carrying pTef1-BbTps3, pGAP-ERG20, pTef1-ERG12 from Yarrowia lipolytica. The plasmid can be effectively amplified in E.coli DH5a and can be successfully expressed in Yarrowia lipolytica. At the same time, ERG12 and ERG20 are important enzyme genes in the MVA pathway, and they carry the PTS1 sequence to help the enzyme protein locate in the peroxisome. Therefore, the plasmid plays an important role in the construction of MVA pathway in the peroxisome of Yarrowia lipolytica.
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 6096
Illegal EcoRI site found at 6146
Illegal EcoRI site found at 7108
Illegal XbaI site found at 5583
Illegal SpeI site found at 1468
Illegal PstI site found at 1285
Illegal PstI site found at 2071
Illegal PstI site found at 2826
Illegal PstI site found at 3406
Illegal PstI site found at 3796
Illegal PstI site found at 5209 - 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 23INCOMPATIBLE WITH RFC[23]Unknown
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 6096
Illegal EcoRI site found at 6146
Illegal EcoRI site found at 7108
Illegal XbaI site found at 5583
Illegal SpeI site found at 1468
Illegal PstI site found at 1285
Illegal PstI site found at 2071
Illegal PstI site found at 2826
Illegal PstI site found at 3406
Illegal PstI site found at 3796
Illegal PstI site found at 5209
Illegal NgoMIV site found at 2200
Illegal NgoMIV site found at 3081
Illegal NgoMIV site found at 3772
Illegal NgoMIV site found at 6234
Illegal AgeI site found at 3680
Illegal AgeI site found at 7705 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1966
Illegal BsaI.rc site found at 3872
Illegal BsaI.rc site found at 5826
Illegal SapI site found at 5179