Plasmid
pNW33N+la

Part:BBa_K3842010:Design

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


pNW33N+lac+ Gas6


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 4319
    Illegal XbaI site found at 4186
    Illegal PstI site found at 4174
    Illegal PstI site found at 5264
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4319
    Illegal NheI site found at 6395
    Illegal PstI site found at 4174
    Illegal PstI site found at 5264
    Illegal NotI site found at 6380
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4319
    Illegal BglII site found at 4830
    Illegal BamHI site found at 4192
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 4319
    Illegal XbaI site found at 4186
    Illegal PstI site found at 4174
    Illegal PstI site found at 5264
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 4319
    Illegal XbaI site found at 4186
    Illegal PstI site found at 4174
    Illegal PstI site found at 5264
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 548
    Illegal BsaI.rc site found at 2239
    Illegal SapI.rc site found at 1630


Design Notes

The plasmid is large, when generating, mutations needs more care.


Source

It is generated by combination ofpNW33N plasmide backbone, T7 promoter, lac operator, Gas6 coding sequence, T7 terminator.

References

[1]Yang, C. K., Zhang, X. Z., Lu, C. D., & Tai, P. C. (2014). An internal hydrophobic helical domain of Bacillus subtilis enolase is essential but not sufficient as a non-cleavable signal for its secretion. Biochemical and biophysical research communications, 446(4), 901–905. https://doi.org/10.1016/j.bbrc.2014.03.032


[2]Yang, C. K., Ewis, H. E., Zhang, X., Lu, C. D., Hu, H. J., Pan, Y., Abdelal, A. T., & Tai, P. C. (2011). Nonclassical protein secretion by Bacillus subtilis in the stationary phase is not due to cell lysis. Journal of bacteriology, 193(20), 5607–5615. https://doi.org/10.1128/JB.05897-11


[3]Yang, C. K., Tai, P. C., & Lu, C. D. (2014). Time-related transcriptome analysis of B. subtilis 168 during growth with glucose. Current microbiology, 68(1), 12–20. https://doi.org/10.1007/s00284-013-0432-4


[4]Yang, C. K., Lu, C. D., & Tai, P. C. (2013). Differential expression of secretion machinery during bacterial growth: SecY and SecF decrease while SecA increases during transition from exponential phase to stationary phase. Current microbiology, 67(6), 682–687. https://doi.org/10.1007/s00284-013-0421-7