Coding

Part:BBa_K3815005

Designed by: Shiotsu Koudai   Group: iGEM21_Kyoto   (2021-10-16)
Revision as of 10:40, 20 October 2021 by Shiotsu (Talk | contribs)


NOP1v-Mxe GryA intein-PT-linker-ELK16

Descriotion of this part

Targeted protein

This part is for the purification of NOP1v.This is made by adding a valine to the N-terminus of NOP1.The effect of it is the same as NOP1. We could not get NOP1 sufficiently when using BBa_K3815004, so we added a valine to the N-terminus of NOP1 according to N-end-rule.

Sequence and Features

Purification system

In order to purify the peptide, we adopted ELK16 system that the past iGEM teams had not used. This part is composed of ELK16, Mxe Gyr intein, and PT linker. When this fused protein is produced, ELK16 self-assembles and precipitates. After that, taking this aggregate and adding DTT to it, the N terminal of Mxe GyrA intein is cut. Then, we can get the targeted protein.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 120
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 120
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 120
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 120
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 120
    Illegal NgoMIV site found at 553
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None