Composite

Part:BBa_K4227025:Design

Designed by: Yang Chen   Group: iGEM22_ZJUT-China   (2022-10-13)


PnlmC-muB-sigma28


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 669
    Illegal EcoRI site found at 1638
    Illegal EcoRI site found at 3173
    Illegal PstI site found at 1587
    Illegal PstI site found at 1888
    Illegal PstI site found at 2200
    Illegal PstI site found at 2763
    Illegal PstI site found at 3487
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 669
    Illegal EcoRI site found at 1638
    Illegal EcoRI site found at 3173
    Illegal NheI site found at 1113
    Illegal PstI site found at 1587
    Illegal PstI site found at 1888
    Illegal PstI site found at 2200
    Illegal PstI site found at 2763
    Illegal PstI site found at 3487
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 669
    Illegal EcoRI site found at 1638
    Illegal EcoRI site found at 3173
    Illegal BglII site found at 413
    Illegal BglII site found at 676
    Illegal BglII site found at 2974
    Illegal BglII site found at 3746
    Illegal BamHI site found at 3355
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 669
    Illegal EcoRI site found at 1638
    Illegal EcoRI site found at 3173
    Illegal PstI site found at 1587
    Illegal PstI site found at 1888
    Illegal PstI site found at 2200
    Illegal PstI site found at 2763
    Illegal PstI site found at 3487
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 669
    Illegal EcoRI site found at 1638
    Illegal EcoRI site found at 3173
    Illegal PstI site found at 1587
    Illegal PstI site found at 1888
    Illegal PstI site found at 2200
    Illegal PstI site found at 2763
    Illegal PstI site found at 3487
    Illegal NgoMIV site found at 801
    Illegal NgoMIV site found at 1703
    Illegal NgoMIV site found at 2702
    Illegal NgoMIV site found at 2991
    Illegal NgoMIV site found at 3069
    Illegal NgoMIV site found at 3543
    Illegal NgoMIV site found at 3888
    Illegal AgeI site found at 257
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We design the part as the inhibit module of our oral probiotic. The PnlmC was a kind of inducible promoter of the CSP from the S.mutans. We choose it to better control the expression of the muB and sigma28.

The muB is chosen as a kind of the α-1,3 dextranase to decompose the EPS, and the sigma28 was used to control our kill-switch.

Source

Synthesized by the Tsingke Biotechnology Co., Ltd.

References

[1] Tianlei Liu, Shoubin Xue, Wenbo Cai, Xiaojing Liu, Xiaojuan Liu, Rongrong Zheng, Hongyan Luo, Wenhui Qi. ComCED signal loop precisely regulates nlmC expression in Streptococcus mutans. Annals of Microbiology.2013;1 (64) : 31-38..

[2] R Lutz. Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic Acids Research.1997;6 (25) : 1203-1210.

[3] Nobuyuki Sumitomo, Katsuhisa Saeki, Katsuya Ozaki, Susumu Ito, Tohru Kobayashi. Mutanase from a Paenibacillus isolate: Nucleotide sequence of the gene and properties of recombinant enzymes. Biochimica et Biophysica Acta (BBA) - General Subjects.2007;4 (1770) : 716-724.