Difference between revisions of "Part:BBa K4158010"
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We designed <b>Psrtf-GFP(<partinfo>BBa_K4158010</partinfo>)</b>, and confirmed its activity <i>in vitro</i>. This part is the reporter plasmid used for progesterone detection. It encodes GFP gene in the downstream of the binding site of SRTF1, the transcriptional factor specific to progesterone. | We designed <b>Psrtf-GFP(<partinfo>BBa_K4158010</partinfo>)</b>, and confirmed its activity <i>in vitro</i>. This part is the reporter plasmid used for progesterone detection. It encodes GFP gene in the downstream of the binding site of SRTF1, the transcriptional factor specific to progesterone. | ||
| − | + | Promoter and SRTF1 binding site:We cited information about the sequence of SRTF1 dependent promoter from paper[1]. | |
| − | Consequently, the promoter sequence was the same as <partinfo>BBa_J32102</partsinfo> and the SRTF1 binding site was the same as | + | Consequently, the promoter sequence was the same as <partinfo>BBa_J32102</partsinfo> and the SRTF1 binding site was the same as BBa_K3889030. |
| − | + | RBS:We designed the RBS by using RBS calculator and RNA fold as it optimizes to GFPuv. | |
Then, we constructed this part by infusion cloning. A fragment of Psrtf1-gfp was inserted into pACYC184 vector which was cut on restriction enzyme sites of HindIII and BamHI. | Then, we constructed this part by infusion cloning. A fragment of Psrtf1-gfp was inserted into pACYC184 vector which was cut on restriction enzyme sites of HindIII and BamHI. | ||
Revision as of 11:08, 10 October 2022
Psrtf1-GFP
This part contains RBS, GFPuv coding site and a promoter regulated by SRTF1 and Progesterone((1S,3aS,3bS,9aR,9bS,11aS)-1-Acetyl-9a,11a-dimethyl-1,2,3,3a,3b,4,5,8,9,9a,9b,10,11,11a-tetradecahydro-7H-cyclopenta[a]phenanthren-7-one) and works as the reporter plasmid to confirm exist of Progesterone and SRTF1.
RBS+GFPuv sequence is the same as BBa_K4158011.
We designed Psrtf-GFP(BBa_K4158010), and confirmed its activity in vitro. This part is the reporter plasmid used for progesterone detection. It encodes GFP gene in the downstream of the binding site of SRTF1, the transcriptional factor specific to progesterone.
Promoter and SRTF1 binding site:We cited information about the sequence of SRTF1 dependent promoter from paper[1]. Consequently, the promoter sequence was the same as No part name specified with partinfo tag.) was successfully achieved</b>. we achieved a project based on BioBricks, which is an important standard component in synthetic biology. </li> </ul>
Thus, we succeeded in engineering SRTF1 regulated gfp reporter plasmid(BBa_K4158010).
(For more details, go to https://2022.igem.wiki/waseda-tokyo/results#progesterone-detection.)
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 13
Illegal NheI site found at 36 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 826
Illegal XhoI site found at 529 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
References
1. Sankar K et al. A progesterone biosensor derived from microbial screening. ACS Sens. 7(4):1132-1137(2022).