Plasmid
pPICZ^

Part:BBa_K3842006:Design

Designed by: Xiaoyi Feng   Group: iGEM21_GA_State_SW_Jiaotong   (2021-10-17)


pPICZαA-Gas6(plasmid)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1215
    Illegal EcoRI site found at 4786
    Illegal XbaI site found at 1235
    Illegal PstI site found at 5731
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1215
    Illegal EcoRI site found at 4786
    Illegal PstI site found at 5731
    Illegal NotI site found at 1221
    Illegal NotI site found at 6847
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1215
    Illegal EcoRI site found at 4786
    Illegal BglII site found at 1
    Illegal BglII site found at 3553
    Illegal BglII site found at 5297
    Illegal BamHI site found at 1641
    Illegal XhoI site found at 1191
    Illegal XhoI site found at 4754
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1215
    Illegal EcoRI site found at 4786
    Illegal XbaI site found at 1235
    Illegal PstI site found at 5731
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1215
    Illegal EcoRI site found at 4786
    Illegal XbaI site found at 1235
    Illegal PstI site found at 5731
    Illegal NgoMIV site found at 2399
    Illegal NgoMIV site found at 2460
    Illegal AgeI site found at 1360
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1624
    Illegal BsaI.rc site found at 1823
    Illegal BsaI.rc site found at 2821


Design Notes

I have to consider the position of coding sequence, it must between the promoter and terminator. Also, the original plasmid has to been choosen wisely, becasue it's always better to get a high copy plasmid. Finally, the coding sequence of Gas6 needs to be optimized for pichia pastoris.


Source

It was synthesized by a genomic company called Qinke. The sequence was generally edited by inserting Gas6 coding sequence into the gap between AOX1 promoter and AOX1 terminator.

References