Translational_Unit

Part:BBa_K4879011:Design

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


ScRAD52 Expression Construct


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 1780
    Illegal suffix found in sequence at 2050
    Illegal EcoRI site found at 2044
    Illegal SpeI site found at 192
    Illegal PstI site found at 157
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1780
    Illegal EcoRI site found at 2044
    Illegal SpeI site found at 192
    Illegal SpeI site found at 2051
    Illegal PstI site found at 157
    Illegal PstI site found at 2065
    Illegal NotI site found at 1786
    Illegal NotI site found at 2058
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1780
    Illegal EcoRI site found at 2044
    Illegal BglII site found at 766
    Illegal BglII site found at 896
    Illegal XhoI site found at 1586
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 1780
    Illegal suffix found in sequence at 2051
    Illegal EcoRI site found at 2044
    Illegal SpeI site found at 192
    Illegal PstI site found at 157
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 1780
    Illegal EcoRI site found at 2044
    Illegal XbaI site found at 1795
    Illegal SpeI site found at 192
    Illegal SpeI site found at 2051
    Illegal PstI site found at 157
    Illegal PstI site found at 2065
    Illegal NgoMIV site found at 1570
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The expression cassette contained a downstream 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 5'overhang to the guaB 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