Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Yu-chang Ku, Pang-ling Huang
Date created: 2016-10-18 11:00:00
Date modified: 2016-10-20 02:29:35
pSB1C3-LacI inducible Hemagglutinin(H1N1)
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K1955008_sequence (Version 1) |
Description
In order to prove the HA sequence which we designed can smoothly express protein, we use BL21 competent cell to express the HA sequence and detect the protein by Western blot analysis.Notes
Use the standard parts provided by iGEM, we choose BBa_J04500 as a promoter to induce production of protein. BBa_J04500 is a LacI inducible promoter with RBS, so it can be induced by IPTG to open its promoter and start transcription. Double restriction enzyme cutting(EcoRI and XbaI) of pSB1C3-HA to produce a sticky end in the anterior, then also double restriction enzyme cutting(EcoRI and SpeI) of BBa_J04500 to produce other sticky end which can be ligated with EcoRI and XbaI sticky end. Moreover, it will form a scar when XbaI ligate with SpeI, so we can simply compose the pSB1C3-J04500-HA.Source
LacI inducible promoter came from BBa_J04500; hemagglutinin came from BBa_K1955003.| Access | Instance | Definition |
| public public | BBa_J04500 BBa_K1955000 | BBa_J04500 BBa_K1955000 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_J04500 BBa_K1955000 | 1,220 229,1943 | BBa_J04500 BBa_K1955000 |