Part:BBa_K4214003

hGIF with LacI promotor and 6xHis-tag

LacI (BBa_J04500) and Human GIF with His6-tag. This part is a composite part from BBa_K4214002 with the addition of LacI sequence (BBa_J04500). The LacI sequence is used in order to induce our protein expression with IPTG, and the 6xHis-tag was used for protein purification.

Fig.: LacI (BBa_J04500) and Human GIF with His6-tag (BBa_K4214002)

Usage and Biology

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• 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

Characterization

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SDS-PAGE

A production culture was set up using BL21 E. coli and induced with 0.5 mM IPTG once their ODs reached 0.5. The cultures were left overnight at 30°C for protein production. The cells were then harvested, pelleted and purified using a HisPur Cobalt resin. The SDS-PAGE analysis of the production process is shown on the right.

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
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 485
  Illegal BsaI.rc site found at 1165
  Illegal SapI site found at 1155