Difference between revisions of "Part:BBa K4231018"
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Using pUC19 as plasmid backbone, AmpR as reporter gene in bacteria, URA3 tag as reporter gene in Yarrowia lipolytica, shuttle plasmid carrying IDI, NADH-HMG, ERG8 and ERG19 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, IDI, NADH-HMG, ERG8 and ERG19 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, URA3 tag as reporter gene in Yarrowia lipolytica, shuttle plasmid carrying IDI, NADH-HMG, ERG8 and ERG19 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, IDI, NADH-HMG, ERG8 and ERG19 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-IDI-NADH-HMG-URA3-ERG8-ERG19.png | border | center | 400px]] | |
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===Usage and Biology=== | ===Usage and Biology=== |
Latest revision as of 14:26, 7 October 2022
pUC19 carrying with IDI, NADH-HMG, ERG8, ERG19 and URA3 tag from Y.lipolytica
Using pUC19 as plasmid backbone, AmpR as reporter gene in bacteria, URA3 tag as reporter gene in Yarrowia lipolytica, shuttle plasmid carrying IDI, NADH-HMG, ERG8 and ERG19 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, IDI, NADH-HMG, ERG8 and ERG19 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]Unknown
- 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 23INCOMPATIBLE WITH RFC[23]Unknown
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 2325
Illegal EcoRI site found at 8712
Illegal PstI site found at 862
Illegal PstI site found at 1992
Illegal PstI site found at 7315
Illegal NgoMIV site found at 638
Illegal NgoMIV site found at 1366
Illegal NgoMIV site found at 2247
Illegal NgoMIV site found at 2463
Illegal NgoMIV site found at 2967
Illegal NgoMIV site found at 3219
Illegal NgoMIV site found at 3531
Illegal AgeI site found at 2343
Illegal AgeI site found at 3540
Illegal AgeI site found at 3627
Illegal AgeI site found at 4153
Illegal AgeI site found at 8270 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 7991
Illegal BsaI.rc site found at 2386
Illegal BsaI.rc site found at 3048
Illegal BsaI.rc site found at 3396
Illegal BsaI.rc site found at 7710
Illegal BsaI.rc site found at 8060