Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Pai Li
Date created: 2015-09-09 11:00:00
Date modified: 2015-09-10 12:01:46
Fusion protein of Dre::EGFP::ssra-tag, with RBS at beginning
| Types | DnaRegion |
| Roles | Translational_Unit engineered_region |
| Sequences | BBa_K1641008_sequence (Version 1) |
Description
This is a sequence of RBS (BBa_B0034) followed by a fusion protein of Dre::EGFP::ssra-tag. The part is for expression of Dre::EGFP::ssra-tag, which aims to overturn our invertase modules in our Micro-timer.Dre (BBa_K1641002) in our bricks is a Cre-like recombinase with invertase activity that turns the sequence between two anti-parallel VloxP sites up-side-down. An EGFP fusion at the C-term of Dre will not significantly interfere the enzymatic activity, and a flexible chain of 8 AA is added between the two parts to further avoid side-effect like misfolding. An ssra-tag (BBa_M0051) can accelerate the degradation of the protein, in order to clean it up when not in need and reduce the leakage expression.
Note: The brick sequence starts directly after the XbaI of prefix and ends before SpeI of the Suffix, without the useless bases (G and T respectively) added before and after brick sequence. This deletion do not interfere the usage of the brick.
Notes
Ssra-Tag: for efficiently cleaning the protein when not under induction, and helps to reduce leakage expression.Dre is a Cre-like recombinase, so C-term EGFP fusion will not significant reduce its activity.
Source
Dre from BBa_K1641002 and EGFP::ssra-tag from pBI121-EGFP is fused by overlap-PCR; others through connection of bricks.| Sequence Annotation | Location | Component / Role(s) |
| B0034 Dre (in fusion) Flexible chain EGFP-ssra (in fusion) | 1,12 19,1044 1045,1068 1069,1827 | feature/rbs ribosome_entry_site CDS feature/protein sequence_feature feature/misc CDS feature/protein |