Part:BBa_K1189037:Design
His tag + Human ferritin di-subunit with E coil under LacI promoter
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1307
Design notes
This part was created by fusing the heavy chain (BBa_K1189025, P02794 [UniParc]) and light chains (BBa_K1189024, P02792 [UniParc]) of human ferritin together. It is expressed under the LacI promoter (BBa_J04500) and has a his-tag for protein purification. An E-coil (BBa_K1189011) is included in order to allow binding of parts containing the respective K-coil (BBa_K1189010). This particular part is expressed under a strong RBS and the LacI promoter. The iGEM Calgary 2013 team attempted expression and purification of E coil ferritin di-subunit using this system. Additionally, they tested this same coding sequence under the T7 promoter in a different vector.
Source
We synthesized heavy ferrtin (BBa_K1189019) and light ferritin (BBa_K1189020) from a commerical synthesis company. These parts were optimized for expression in E. coli K12. These two elements were extracted via PCR, the pSB1C3 backbone was isolated with PCR, and these linear fragments were assembled into BBa_K1189037 using isothermal assembly.