Component | Description | Part/Accession # | Base Pairs | Plasmid | Resistance | Well |
RBS | Ribosomal Binding Site | BBa_B0034 | 12 | pSB1A2 | Ampicillin | plate 1, 2M |
Cph8 | Red Light sensor | BBa_I15010 | 2,238 | pSB2K3 | Kanamycin | N/A |
RFP | Red Fluorescent Protein | BBa_J04051 | 720 | N/A | ||
OmpR (E. coli) | description | NP_417864.1 | 720 | pSB1T3 | Tetracycline | N/A |
Terminator | Stops Transcription | BBa_B0015 | 129 | pSB1AK3 | Ampicillin and Kanamycin | plate 1, 23L |
OmpR + Terminator | description | sequence | 916 | pany-amp | Ampicillin | synthesized |
OmpC promoter | description | BBa_R0082 | 108 | pSB1A2 | Ampicillin | plate 1, 16K |
puc B/A | description | sequence | 375 | pSB1A3 | Ampicillin | N/A |
puc B | description | YP_353390 | 156 | ? | ? | N/A |
puc A | description | YP_353391 | 165 | ? | ? | N/A |
OmpC promoter+BA | description | sequence | 539 | pany-kana | Kanamycin | synthesized |
Light Response System | description | BBa_M30109 | 4,333 | ? | Ampicillin | N/A |
TetR repressible | description | BBa_J13002 | 74 | pSB1A2 | Ampicillin | plate 1, 13B |
Green Fluorescent Protein | Marker for characterization | BBa_E0240 | 876 | pSB1A2 | Ampicillin | plate 1, 12M |
Plasmids
Plasmid | Description | Base Pairs | Resistance | Copy Number |
pSB1A2 | BioBrick Vector | 2,079 | Ampicillin | high |
pSB1K3 | 2,206 | Kanamycin | high | |
pSB1A3 | Assembly plasmid | 2,157 | Ampicillin | high |
pSB2K3 | 4,425 | Kanamycin | variable | |
pSB1T3 | Assembly plasmid | 2,463 | Tetracycline | high |
pSB1AK3 | Assembly Plasmid | 3,189 | Ampicillin and Kanamycin | high |
pANY | Synthesized from Geneart | |||
pRKCBC3 | Tetracycline |
The first part of our characterization begins with the puc promoter. The puc promoter is what turns on the entire system naturally in Rhodobacter sphaeroides. The puc promoter is what ultimately controls the number of LH2 light harvesting complexes, which is how our system will yield an increase in photosynthetic efficiency. It is important that we are able to compare the transcription rate of the puc promoter in the two systems so that we can determine exactly how much efficiency is gained by adding a red light sensor. By attaching Green Fluorescent Protein (GFP) to the promoter we can quantify the rate of transcription by measuring the emission of green light using a fluorescence spectrophotometer. We would expect to see more fluorescence with more transcription and vis versa.
Under Development