Translational_Unit

Part:BBa_K4879010:Design

Designed by: Apoorva Gopal   Group: iGEM23_IISER-Pune-India   (2023-10-08)


Yarrowia lipolytica guaB construct


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 1831
    Illegal suffix found in sequence at 2101
    Illegal EcoRI site found at 928
    Illegal EcoRI site found at 1382
    Illegal EcoRI site found at 2095
    Illegal SpeI site found at 192
    Illegal PstI site found at 157
    Illegal PstI site found at 1241
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 928
    Illegal EcoRI site found at 1382
    Illegal EcoRI site found at 1831
    Illegal EcoRI site found at 2095
    Illegal SpeI site found at 192
    Illegal SpeI site found at 2102
    Illegal PstI site found at 157
    Illegal PstI site found at 1241
    Illegal PstI site found at 2116
    Illegal NotI site found at 1837
    Illegal NotI site found at 2109
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 928
    Illegal EcoRI site found at 1382
    Illegal EcoRI site found at 1831
    Illegal EcoRI site found at 2095
    Illegal BamHI site found at 1444
    Illegal XhoI site found at 1214
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 1831
    Illegal suffix found in sequence at 2102
    Illegal EcoRI site found at 928
    Illegal EcoRI site found at 1382
    Illegal EcoRI site found at 2095
    Illegal SpeI site found at 192
    Illegal PstI site found at 157
    Illegal PstI site found at 1241
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 1831
    Illegal EcoRI site found at 928
    Illegal EcoRI site found at 1382
    Illegal EcoRI site found at 2095
    Illegal XbaI site found at 1846
    Illegal SpeI site found at 192
    Illegal SpeI site found at 2102
    Illegal PstI site found at 157
    Illegal PstI site found at 1241
    Illegal PstI site found at 2116
    Illegal NgoMIV site found at 1231
    Illegal NgoMIV site found at 1456
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The expression cassette contained an upstream sequence homologous to the A08 gene locus, as it was previously seen that gene insertion by recombination at this locus caused very little detrimental effect on the chassis (Qingchun Ji et al. 2020). There is also a 3'overhang to the ScRAD52 expression construct in order to facilitate DNA assembly.


Source

The expression construct was synthesised by IDT.


References

Qingchun Ji, Jie Mai, Ying Ding, Yongjun Wei, Rodrigo Ledesma-Amaro, Xiao-Jun Ji, Improving the homologous recombination efficiency of Yarrowia lipolytica by grafting heterologous component from Saccharomyces cerevisiae, Metabolic Engineering Communications, Volume 11, 2020, e00152, ISSN 2214-0301, https://doi.org/10.1016/j.mec.2020.e00152