Parts

Introduction

Welcome to the page where you can find all the parts created by iGEM Stockholm during 2022–2023. These parts are present and documented on the Registry of Standard Biological Parts.

BBa_K4214000

This part codes for human gastric intrinsic factor protein (AA19-417). The gene is located in chromosome 11q12.1. This gene is a part of cobalamin transporters group. It is essential for the absorption of vitamin B12. Mutations in this gene may lead to pernicious anaemia.

We removed the native signal peptide (AA1-18) from the N-terminal in order to obtain a sequence that can be worked with more easily in bacterial and mammalian systems. Since we used a surface display expression vector which already includes an IgK leader sequence and other flanking sequences, we did not need the GIF signal sequence.

BBa_K4214002

Human GIF with His6-tag. This part has the native signal peptide removed, as it is not required for bacterial expression or in our mammalian expression vector. This part was used to produce and purify our Human GIF in bacteria, through the composite parts BBa_K4214003 (with LacI promotor) and BBa_K4214004 (with LacI promotor and pelB leader sequence).

BBa_K4214003

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.

BBa_K4214004

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 transport our protein to 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.