Composite

Part:BBa_K3239011:Design

Designed by: Xingyu Zhou   Group: iGEM19_ShanghaiFLS_China   (2019-10-16)


pMIT1-PRM1-pPRM1-MIT1


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2034
    Illegal SpeI site found at 793
    Illegal PstI site found at 2556
    Illegal PstI site found at 6970
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2034
    Illegal NheI site found at 3815
    Illegal SpeI site found at 793
    Illegal PstI site found at 2556
    Illegal PstI site found at 6970
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2034
    Illegal BglII site found at 1387
    Illegal BglII site found at 4268
    Illegal BglII site found at 7428
    Illegal BamHI site found at 2917
    Illegal BamHI site found at 3836
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2034
    Illegal SpeI site found at 793
    Illegal PstI site found at 2556
    Illegal PstI site found at 6970
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2034
    Illegal SpeI site found at 793
    Illegal PstI site found at 2556
    Illegal PstI site found at 6970
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The two subparts shall be separately integrated into the PAOX1-GFP strain. They are therefore not necessarily conjoined as shown in the sequence illustration.

Source

P.pastoris GS115 and https://www.ncbi.nlm.nih.gov/nuccore/82569479

Registered parts BBa_K3239010, BBa_K3239009

References

yEGFP3 protein: https://www.ncbi.nlm.nih.gov/protein/82569482

yEGFP2 sequence source: https://www.ncbi.nlm.nih.gov/nuccore/82569479

Liang, S., Wang, B., Pan, L., Ye, Y., He, M., Han, S., … Lin, Y. (2012). Comprehensive structural annotation of Pichia pastoris transcriptome and the response to various carbon sources using deep paired-end RNA sequencing. BMC Genomics, 13(1). https://doi.org/10.1186/1471-2164-13-738

Shi, L., Wang, J., Wang, X., Zhang, Y., Song, Z., Cai, M., & Zhou, X. (2019). Transcriptome and metabolome analyses reveal global behaviour of a genetically engineered methanol-independent Pichia pastoris strain. Process Biochemistry, 76(August 2018), 46–54. https://doi.org/10.1016/j.procbio.2018.10.014

Wang, X., Wang, Q., Wang, J., Zhou, M., Shi, L., Zhou, X., … Shen, W. (2016). Mit1 Transcription Factor Mediates Methanol Signaling and Regulates the Alcohol Oxidase 1 ( AOX1 ) Promoter in Pichia pastoris. Journal of Biological Chemistry, 291(12), 6245–6261. https://doi.org/10.1074/jbc.m115.692053