Part:BBa_K4390016

6xHis :: TEV protease cleavage site

This part is not compatible with BioBrick RFC10 assembly but is compatible with the iGEM Type IIS Part standard which is also accepted by iGEM.

This is a Level 0 part of type N part generating the following 4 base overhangs at upstream (AATG) and downstream (AGCC) ends.

Usage and Biology

The TEV protease comes from the Tobacco Etch Virus (TEV). It is a cysteine protease that is commonly used as it has high specificity to particular sequences. The 7-amino acid sequence is commonly used as it is best recognised by this TEV protease, which belongs to the family of chymotrypsin-like proteases. By inserting this His-TEV Protease cleavage site, you are able to cleave the fusion protein directly in the solution or whilst it is immobilized on affinity resins. It can be used for the removal of tags on recombinant proteins both in vitro and in vivo. The optimum temperature for cleavage by the TEV protease of this characteristic sequence is at 30°C.

This part fuses the 6-His tag with the TEV cutting site. Thus fusion of protein with this part can be purified using nickle colomn and the His-tag can be cleaved by TEV protease after purification to remove any possibility of influence from His-tag.

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
  COMPATIBLE WITH RFC[1000]