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 07:14:11
Fusion protein of Cre-EGFP, with RBS at the beginning
| Types | DnaRegion |
| Roles | Translational_Unit engineered_region |
| Sequences | BBa_K1641042_sequence (Version 1) |
Description
This is a sequence of RBS (BBa_B0034) followed by a fusion protein of Cre-EGFP. The part is for expression of Cre-EGFP, which aims to overturn our invertase modules in our Micro-timer.Cre (BBa_K1179058) is a most commonly used recombinase with invertase activity that turns the sequence between two anti-parallel LoxP sites up-side-down. An EGFP fusion at the C-term of Vcre 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. Unlike other invertase module in Micro-timer of SYSU-CHINA in 2015, this part does not have ssra-tag, and thus has better expression efficiency and enzymatic activity.
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. The scar will be "ACTAGA" safely without start code and restriction enzyme activity. This sequence is absolutely compatible with brick construction method and usually less likely to make trouble.
Notes
This Cre-EGFP do not process ssra, and hence much stable in vivo. As a result, the enzymatic activity seems much higher than those with ssra. Yet, a tiny leakage express might happen even if linked to inductive promoter.Source
Cre from BBa_K1179058 and EGFP from pBI121-EGFP is fused by overlap-PCR; RBS is added via PCR primer.| Sequence Annotation | Location | Component / Role(s) |
| B0034 Cre (in fusion) flexible chain EGFP (in fusion) | 1,12 19,960 961,984 985,1704 | feature/rbs ribosome_entry_site feature/protein CDS sequence_feature feature/misc feature/protein CDS |