Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Pai Li
Date created: 2015-09-08 11:00:00
Date modified: 2015-09-10 03:02:39
Fusion protein of Flp-EGFP-ssra, with RBS at beginning
| Types | DnaRegion |
| Roles | engineered_region Translational_Unit |
| Sequences | BBa_K1641005_sequence (Version 1) |
Description
This is a sequence of RBS (BBa_B0034) followed by a fusion protein of Flp::EGFP::ssra-tag. This part is for expression of Flp::EGFP::ssra-tag, which aims to overturn our invertase modules in our Micro-timer.Flp is a commonly used recombinase originated from yeast, with invertase activity that turns the DNA sequence between two anti-parallel FRT sites up-side-down. A EGFP fusion at the C-term of Flp will not significantly interfere the enzymatic activity. A ssra-tag (BBa_M0051) can accelerate the degradation of the protein, in order to clean it up when not in need.
Notes
Ssra-Tag is efficiently cleaning the protein when not under induction, and helps to reduce leakage expression. C-term EGFP fusion on Flp seems have far better activity than N-term fusion.Source
Flp is from BBa_K1015001 and EGFP::ssra-tag from pBI121-EGFP in Yongjun Lu's lab is fused by overlap-PCR; others through connection of bricks.| Sequence Annotation | Location | Component / Role(s) |
| B0034 scar flexible chain Flp (without stop) EGFP-ssra(M0051) | 1,12 13,18 1287,1311 19,1286 1312,2070 | feature/rbs ribosome_entry_site feature/scar sequence_feature feature/misc CDS feature/protein CDS feature/protein |