Coding

Part:BBa_K4214004

Designed by: Jol Tekoniemi   Group: iGEM22_Stockholm   (2022-10-08)


hGIF with LacI promotor, pelB sequence and 6xHis-tag

LacI (BBa_J04500) and pelB sequence (BBa_J32015) and Human GIF with His6-tag. This part is a composite part from BBa_K4214002 with the addition of pelB sequence (BBa_J32015) and LacI (BBa_J04500). The pelB sequence is used in order to express our protein in the periplasm. The enzymes in the periplasm of E.coli can add disulfide bonds in our protein, ensuring its correct conformation. This part was used to produce and purify our Human GIF.


Fig.: LacI(BBa_J04500), pelB sequence (BBa_J32015) and Human GIF with His6-tag (BBa_K4214002)


Usage and Biology

  • NCBI page of the human gastric intrinsic factor [1]
  • This part of hGIF does not contain the native signal peptide (AA1-18)
  • The LacI promotor is described in part BBa_J04500
  • The pelB sequence is described in the part BBa_J32015


Characterization


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 280
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 557
    Illegal BsaI.rc site found at 1237
    Illegal SapI site found at 1227


[edit]
Categories
Parameters
None