Difference between revisions of "Part:BBa K2680050"
(→Design) |
|||
| (One intermediate revision by the same user not shown) | |||
| Line 5: | Line 5: | ||
===Design=== | ===Design=== | ||
| − | This part is a j23107 ts-cI repressed mf-Lon. For more information on mf-Lon, and to see the source of the mf-Lon, see BBa_K2333011 | + | This part is a j23107 ts-cI repressed mf-Lon. For more information on mf-Lon, and to see the source of the mf-Lon, see <partinfo>BBa_K2333011</partinfo> |
===Mechanics=== | ===Mechanics=== | ||
| Line 14: | Line 14: | ||
Mf-Lon is capable of degrading proteins tagged with a specific pdt tag. If used in combination with such a tagged protein, an IFFL circuit can be produced. During our research, we utilized BBa_K2680050 as our initial heat sensitive mf-Lon. After experimentation, we found that the amount of mf-Lon produced was too high to create an IFFL circuit, as either not enough mscarlet-I could be produced to generate a pulse, or the amount of mf-Lon and mscarlet-I was too high, causing cell death (see the figure below). | Mf-Lon is capable of degrading proteins tagged with a specific pdt tag. If used in combination with such a tagged protein, an IFFL circuit can be produced. During our research, we utilized BBa_K2680050 as our initial heat sensitive mf-Lon. After experimentation, we found that the amount of mf-Lon produced was too high to create an IFFL circuit, as either not enough mscarlet-I could be produced to generate a pulse, or the amount of mf-Lon and mscarlet-I was too high, causing cell death (see the figure below). | ||
| − | "https://static.igem.org/mediawiki/2018/ | + | "https://static.igem.org/mediawiki/2018/e/ea/T--William_and_Mary--014_%2B_012_death.png" |
Latest revision as of 00:48, 18 October 2018
J23107 ts-CI+ R0051 mf-Lon
Design
This part is a j23107 ts-cI repressed mf-Lon. For more information on mf-Lon, and to see the source of the mf-Lon, see BBa_K2333011
Mechanics
In this circuit, temperature-sensitive CI is expressed by J23107, a medium-weak constitutive promoter. This temperature-sensitive CI then represses the downstream CI promoter. Once the temperature is sufficiently raised, the ts-CI protein denatures, loosing its repression capabilities. Therefore, the CI promoter is freed, allowing for expression of mf-Lon.
Characterization
Mf-Lon is capable of degrading proteins tagged with a specific pdt tag. If used in combination with such a tagged protein, an IFFL circuit can be produced. During our research, we utilized BBa_K2680050 as our initial heat sensitive mf-Lon. After experimentation, we found that the amount of mf-Lon produced was too high to create an IFFL circuit, as either not enough mscarlet-I could be produced to generate a pulse, or the amount of mf-Lon and mscarlet-I was too high, causing cell death (see the figure below).
"
"
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 51
Illegal NheI site found at 74 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 2903
Illegal AgeI site found at 2987
Illegal AgeI site found at 3193
Illegal AgeI site found at 3218 - 1000COMPATIBLE WITH RFC[1000]