Plasmid

Part:BBa_K4231019

Designed by: Shi Han   Group: iGEM22_USTC   (2022-09-30)


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-Bs, 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.

PUC-BbTps3-ERG12-ERG20-LEU.png


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE 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
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 6096
    Illegal EcoRI site found at 6146
    Illegal EcoRI site found at 7108
    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
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6096
    Illegal EcoRI site found at 6146
    Illegal EcoRI site found at 7108
    Illegal BglII site found at 812
    Illegal BglII site found at 944
    Illegal BglII site found at 8377
    Illegal BamHI site found at 2447
    Illegal BamHI site found at 5528
    Illegal XhoI site found at 1073
    Illegal XhoI site found at 1229
    Illegal XhoI site found at 3979
    Illegal XhoI site found at 7091
    Illegal XhoI site found at 7120
  • 23
    INCOMPATIBLE WITH RFC[23]
    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
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE 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


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Categories
Parameters
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