Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Fredrik Lindeberg
Date created: 2015-09-14 11:00:00
Date modified: 2015-09-15 01:47:52
ModLac laccase with His-tag and HlyA export tag
| Types | DnaRegion |
| Roles | CDS Coding |
| Sequences | BBa_K1688020_sequence (Version 1) |
Description
A modified laccase (D439A/M510L CueO) with N-His6 tag attached to it via a linker sequence and an export tag (HlyA, BBa_K554002) attached to its C-terminus.Notes
The modified CueO was synthesized with iGEM prefix and suffix, RBS(BBa_B0034) and a M-scar after the RBS in order to guarantee the function of the RBS. Two Nsil restriction sites were put before and after a 6xHis-tag so that the enzyme could be purified with an IMAC and so that the tag could be removed. The gene is followed by a KpnI restriction site with glycines and serines on each side, which are small amino acids which increase the flexibility of the protein. This is needed for the export tag HlyA , to make it easier for the resulting signal protein to be folded properly. A second KpnI restriction site is put after the HlyA so that the export tag can be removed easily. It has then two stop codons, and a terminator at the end.Source
D439A/M510L CueO mutant sequence received from Professor Kunishige Kataoka and synthesized with His-tag and HlyA export tag.| Sequence Annotation | Location | Component / Role(s) |
| 6xHis HlyA export tag | 4,20 1590,1781 | tag feature/tag tag feature/tag |