DNA

Part:BBa_K4765135

Designed by: Siliang Zhan   Group: iGEM23_Fudan   (2023-10-10)


ribozyme connected: Dsup + H. ex mtSSB + SAHS 33020 + XRCC1 + FEN1

contributed by Fudan iGEM 2023

Usage and Biology

This part is the combination of all the anti-desiccation and anti-UV proteins, we've transformed it to E .coli BL21 DE3 and test its performance.

Characterization

Agarose gel electrophoresis

contributed by Fudan iGEM 2023
Figure 1. Agarose gel electrophoresis of PCR products, amplified from bacterial colonies/cultures.

From right lane(3) to left lane(1) indicate the successful construction of Dsup, Dsup + H. ex mtSSB, and Dsup + H. ex mtSSB + SAHS 33020.

Successful Protein Expression

contributed by Fudan iGEM 2023
Figure 2. SDS-PAGE electrophoresis of BBa K4765135

We constructed BBa K4765135 into the pET28a plasmid and transformed it into E. coli BL21 DE3. This lane represents the leaky expression of BBa K4765135. As indicated by the red arrow, we successfully expressed Rv Dsup, H. ex mtSSB, SAHS 33020 and XRCC1 in this part.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 3307
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 5485
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1516
    Illegal NgoMIV site found at 6063
    Illegal AgeI site found at 230
    Illegal AgeI site found at 301
    Illegal AgeI site found at 5169
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2149
    Illegal BsaI.rc site found at 227
    Illegal BsaI.rc site found at 1784
    Illegal BsaI.rc site found at 1865
    Illegal BsaI.rc site found at 4357
    Illegal SapI site found at 1678
    Illegal SapI.rc site found at 956


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