Coding

Part:BBa_K2881000

Designed by: Zhijian Li, Ruiyuan Zhang   Group: iGEM18_UC_San_Diego   (2018-10-04)


mMBD-eGFP-avi-His

MBD stands for Methyl-CpG-binding proteins. This family of proteins contain MBD domains specifically binding to methylated CpG sites in vitro. Our part aims to utilize MBD domain to quantify methylation levels of circulating tumor DNA (ctDNA); it is also useful for characterizing other methylated DNA substrates. A single mMBD domain is fused to eGFP to provide a way to visualize the signal; eGFP also significantly enhances expression of this part in BL-21(DE3). His-tag enables the protein to be purified by Nickel column. Avi-tag serves as a versatile platform for other signal detection/amplification possibilities: it could be specifically recognized by BirA enzyme in vitro to attach a biotin.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 201
    Illegal BamHI site found at 1287
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 2080


[edit]
Categories
Parameters
None