Parts are essential to the engineering component of synthetic biology. With the registry of parts, we can standardize biological parts and share data to advance our progress as a scientific community.
This year, the Penn State iGEM Team has produced a number of new riboswitch sequences and associated parts. Our riboswitches serve as an excellent example of the modular applications of biological parts; unfortunately, unexpected setbacks delayed our characterizations. Listed below are the basic parts, composite parts, and collections of parts that we have designed and hope better characterize in the future. Riboswitch parts were only inputted for mCRP, bFGF, and BThrom riboswitches with the intent of collecting more data for these parts before the Jamboree.
Our basic parts consist of each component of our SensoREX plasmid. These parts include a variety of riboswitch designs to test in different combinations with either the J23100 promoter or the T7 promoter in the pFTV1 vector.
1. pFTV1 Vector: BBa_K4950000
2. mRFP1 spacer + Tre37A: BBa_K4950001
3. mCRP1 Riboswitch Insert: BBa_K4950002
4. mCRP2 Riboswitch Insert: BBa_K4950003
5. mCRP3 Riboswitch Insert: BBa_K4950004
6. mCRP4 Riboswitch Insert: BBa_K4950005
7. bFGF1 Riboswitch Insert: BBa_K4950007
8. mCRP5 Riboswitch Insert: BBa_K4950006
9. bFGF2 Riboswitch Insert: BBa_K4950008
10. bFGF3 Riboswitch Insert: BBa_K4950009
11. bFGF4 Riboswitch Insert: BBa_K4950010
12. bFGF5 Riboswitch Insert: BBa_K4950011
13. bFGF5 Riboswitch Insert: BBa_K4950011
14. BThrom1 Riboswitch Insert: BBa_K4950012
15. BThrom2 Riboswitch Insert: BBa_K4950013
16. BThrom3 Riboswitch Insert: BBa_K4950014
17. BThrom4 Riboswitch Insert: BBa_K4950015
18. BThrom5 Riboswitch Insert: BBa_K4950016
19. BThrom5 Riboswitch Insert: BBa_K4950016
20. mRFP1 Codon Optimized: BBa_K4950017
21. pFTV1 T7 Insert: BBa_K4950018
22. pFTV1 J23100 Insert: BBa_K4950019
The composite parts are composed of our complete plasmids with pFTV1 vector, J23100 promoter, ON riboswitch, and mRFP1 gene. In the presence of the biomarker, the riboswitch mRNA is intended to undergo a conformational change to expose the ribosome binding site, permitting translation of the downstream mRFP1 gene.
1. pFTV1_mCRP1_mRFP1 Riboswitch: BBa_K4950020
2. pFTV1_mCRP2_mRFP1 Riboswitch: BBa_K4950021
3. pFTV1_mCRP3_mRFP1 Riboswitch: BBa_K4950022
4. pFTV1_mCRP4_mRFP1 Riboswitch: BBa_K4950023
5. pFTV1_mCRP5_mRFP1 Riboswitch: BBa_K4950024
6. pFTV1_bFGF1_mRFP1 Riboswitch: BBa_K4950025
7. pFTV1_bFGF2_mRFP1 Riboswitch: BBa_K4950026
8. pFTV1_bFGF3_mRFP1 Riboswitch: BBa_K4950027
9. pFTV1_bFGF4_mRFP1 Riboswitch: BBa_K4950028
10. pFTV1_bFGF5_mRFP1 Riboswitch: BBa_K4950029
11. pFTV1_BThrom1_mRFP1 Riboswitch: BBa_K4950030
12. pFTV1_BThrom2_mRFP1 Riboswitch: BBa_K4950031
13. pFTV1_BThrom3_mRFP1 Riboswitch: BBa_K4950032
14. pFTV1_BThrom4_mRFP1 Riboswitch: BBa_K4950033
15. pFTV1_BThrom5_mRFP1 Riboswitch: BBa_K4950034
Our collection of parts serves as a sample template for the SensoREX system. This collection includes the vector, both the J23100 and T7 promoter inserts, the BThrom4 ON riboswitch, and both the mRFP1 and modified Tre37A coding sequences. The BThrom4 riboswitch was included in the collection as it is one of our more promising designs, and we plan to better characterize the part so that other teams may use it as a standard. This collection holds the components of a modular system; as more RNA aptamers are discovered, future teams may implement their own designs into our plasmid and can mix and match as they please.
1. pFTV1 Vector: BBa_K4950000
2. mRFP1 spacer + Tre37A: BBa_K4950001
3. BThrom4 Riboswitch Insert: BBa_K4950015
4. mRFP1 Codon Optimized: BBa_K4950017
5. pFTV1 T7 Insert: BBa_K4950018
6. pFTV1 J23100 Insert: BBa_K4950019