Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Changxing Hu
Date created: 2015-09-01 11:00:00
Date modified: 2015-09-07 09:04:10
SV40(with Enhancer)+Rluc Composite
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K1722012_sequence (Version 1) |
Description
SV40 is an abbreviation for Simian Virus 40, a polyomavirus that is found in both monkeys and humans. SV40 promoter, which is one of the earliest virus promoter being found by biologists, can improve the gene expression level of many host cells. Similar to 35S promoter, SV40 has a relatively small genetic structure and high expression driving ability. As a strong promoter being widely used in genetic engineering, SV40 has a close affinity with RNA polymerase and can direct the massive synthesizing of mRNA. Different from the SV40 promoter that is already existed in iGEM Distribution kit, this promoter has an enhancer in its sequence.Rluc can express Renilla luciferase which has become popular as a reporter enzyme for gene expression assays. Renilla luciferase(RLUC) is a blue-light emitting luciferase of marine anthozoan Renilla reniformis. As a reporter gene, researchers attach it to a regulatory sequence of another gene of interest in bacteria, cell culture, animals or plants. RLUC is chosen as a reporter because the characteristic it confer on organisms expressing it is easily identified and measured. The bioluminescence of the sea pancy, is under the control of a nerve network and is stimulated by changes of intracellular Ca2+ concerntration. RLUC catalyzes the oxidation of coelenteramide, CO2 and light(480nm）,as in the following scheme.
2015 SZU-iGEM construct SV40 and Rluc with one codon being amber mutated in the same plasmid. This plasmid, together with two other plasmids, are inserted into the cell. Only when the three plasmids work simultanuously can our orthogonal system behave its function, specifically recognise bladder cancer cells and kill them.
Notes
We designed the primers of these two genes and amplified them from the vector psi-Check2, the two genes were then flanked by the iGEM prefix and suffix after amplification. We constructed the two genes in pSB1C3 using 3A Assembly as described in iGEM.org.Source
We achieved both SV40 promoter and Rluc from Shenzhen Second People's Hospital.| Access | Instance | Definition |
| public public | BBa_K1722006 BBa_K1722005 | SV40(en) Rluc |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_K1722006 BBa_K1722005 | 1,419 426,1361 | SV40(en) Rluc |