To be able to gather specific information regarding our project we tested several different constrcuts. Our Best New Composite Part (BBa_K4604023) is made up of three main components; a gene essential for boosted AdoCbl (B12) synthesis, named bluB (BBa_K4604005), a riboswitch as a regulatory element (BBa_K4604031) and a toxin-antitoxin system (BBa_K4604037/BBa_K4604011).
In the following, all of our registered basic and composite parts, along with their functions, are listed.
Basic Parts
| ID | Name | Function |
|---|---|---|
| BBa_K4604002 | TetR repressor | Negatively regulates the tet promoter |
| BBa_K4604003 | rrnB Terminator | Terminates transcription |
| BBa_K4604004 | Modified T7 RBS | Strong RBS, functions as a recruitment area for ribosomes |
| BBa_K4604005 | bluB normal | Gene for BluB, oxidises FMNH2 to form DMB |
| BBa_K4604006 | bluB mutated | Mutated gene for BluB, resulting in premature termination |
| BBa_K4604011 | mazF | Endoribunuclease, cuts cellular RNA |
| BBa_K4604013 | mTurqouise | Fluorescent protein |
| BBa_K4604030 | RBS of piG_01a | Functions as a recruitment area for a ribosome |
| BBa_K4604031 | K12 riboswitch | Negatively regulates gene downstream of it in presence of AdoCbl |
| BBa_K4604032 | PF riboswitch | Negatively regulates downstream genes in presence of AdoCbl |
| BBa_K4604033 | SY riboswitch | Negatively regulates gene downstream of it in presence of AdoCbl |
| BBa_K4604036 | rrnB terminator short | Terminates transcription |
| BBa_K4604037 | mazE | Neutralizes mazF |
| BBa_K4604038 | MazE RBS | Functions as a recruitment area for a ribosome |
| BBa_K4604039 | lacI promoter | Constituitive promoter of the lac-Operon |
| BBa_K4604040 | TetR repressor no sdm | Negatively regulates the tet promoter |
| BBa_K4604043 | terminator of piG_23a/b | Terminates transcription |
| BBa_K4604044 | RBS of AdoCbl riboswitch | Part of the AdoCbl riboswitch, functions as a recruitment area for ribosomes |
| BBa_K4604042 | kanR | Results in kanamycine resistance |
Composite Parts
| ID | Name | Function |
|---|---|---|
| BBa_K4604015 | piG_01b (tetR_bluB) |
Expresses BluB upon induction with doxycycline |
| BBa_K4604016 | piG_01a (leaky_tetR_bluB) |
Expresses BluB upon induction with doxycycline, leaky |
| BBa_K4604017 | piG_02a (leaky_tetR _riboK12_mazF_mTurq) |
Expresses MazF and mTurquoise in little amounts upon induction with doxycycline, leaky, fluorescent |
| BBa_K4604018 | piG_02b (tetR_riboK12 _mazF_mTurq) |
Expresses MazF and mTurquoise in little amounts upon induction with doxycycline, fluorescent |
| BBa_K4604019 | piG_03 (tetR_riboK12_mazF) |
Expresses MazF in little amounts upon induction with doxycycline |
| BBa_K4604020 | piG_07 (tetR_mut_bluB) |
Expresses a mutated, shortened version of BluB upon induction with doxycycline |
| BBa_K4604021 | piG_10a (tetR_bluB _riboK12_mazF) |
Expresses MazF (in absence of AdoCbl) in little amounts and BluB upon induction with doxycycline |
| BBa_K4604022 | piG_10b (tetR_bluB_ T7RBS_riboK12_mazF) |
Expresses MazF (in absence of AdoCbl) and BluB upon induction with doxycycline |
| BBa_K4604023 | piG_21 (tetR_bluB _riboK12_mazF _AmpProm_mazE) |
CELLECT - Regulational unit that should induce cell death in the absence of AdoCbl |
| BBa_K4604024 | piG_23 (tetR_mazF) |
Expressional unit to test MazF toxicity |
| BBa_K4604025 | piG_23b (tetR_RiboRBS_mazF) |
Expressional unit that shows decreased toxicity due to changed RBS |
| BBa_K4604026 | piG_K12BSa (LacIprom _riboK12_lacI _trcProm_sfGFP) |
Expressional unit that makes E. coli glow upon detection of AdoCbl |
| BBa_K4604027 | piG_K12BSb (LacIprom_riboK12 _lacI_trcProm_sfGFP) |
Expressional unit that makes E. coli glow upon detection of AdoCbl, decreased backround fluorescence |
| BBa_K4604028 | piG_PFBS (LacIprom_riboK12 _lacI_trcProm_sfGFP) |
Expressional unit that makes E. coli glow upon detection of AdoCbl |
| BBa_K4604029 | piG_SYBS (LacIprom_riboK12 _lacI_trcProm_sfGFP) |
Expressional unit that makes E. coli glow upon detection of AdoCbl |
| BBa_K4604035 | piG_08 (ampProm_mazE) |
Expresses MazE |
| BBa_K4604041 | piG_11 (tetR_kanR _riboK12_mazF) |
Expresses MazF in little amounts upon induction with doxycycline, added selection marker to ensure transcription of mazF |