Difference between revisions of "Part:BBa K3122003"
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<partinfo>BBa_K3122003 short</partinfo> | <partinfo>BBa_K3122003 short</partinfo> | ||
− | . | + | This part is the constitutive medium promoter, from Anderson family, was adapted to type IIS enzymes standard. |
− | + | It has been used to express our Level 1 constructs related to membrane proteins. The reason behind this choice relays on its relative strength among other members of the family: it is a medium- strength regulator (0.36) which should not express too much membrane protein as to induce cell lysis. | |
− | + | ||
− | < | + | <h2> Characterization </h2> |
− | + | ||
− | + | ||
+ | Characterization of this part has been conducted by building two transcriptional units (BBa_K3122004, BBa_ K3122005), and performing Lambda fague and Cell separation assays. | ||
+ | |||
+ | BBa_K3122004 transcriptional unit was assembled in pARK1 alpha plasmid including the following parts: | ||
+ | <ul> | ||
+ | <li><partinfo>BBa_K3122003</partinfo>: Promoter J23107 adapted to type IIS assembly</li> | ||
+ | <li><partinfo>BBa_K2656009</partinfo>: RBS B0030 adapted to type IIS assembly</li> | ||
+ | <li><partinfo>BBa_K3122002</partinfo>: coding sequence. This composite part is built with BBa_K3122000 (LamB protein) and BBa_K3122001 (6xHis tag) creating our AEGIS' lamB display system exposing 6xHis Tag.</li> | ||
+ | <li><partinfo>BBa_K2656026</partinfo>: Terminator B0015 adapted to type IIS assembly</li> | ||
+ | </ul> | ||
+ | |||
+ | BBa_K3122005 transcriptional unit was assembled in pARK1 alpha plasmid including the following parts: | ||
+ | <ul> | ||
+ | <li><partinfo>BBa_K3122003</partinfo>: Promoter J23107 adapted to type IIS assembly</li> | ||
+ | <li><partinfo>BBa_K2656009</partinfo>: RBS B0030 adapted to type IIS assembly</li> | ||
+ | <li><partinfo>BBa_K3122000</partinfo>: LamB coding sequence. This part is our AEGIS' lamB display system.</li> | ||
+ | <li><partinfo>BBa_K2656026</partinfo>: Terminator B0015 adapted to type IIS assembly</li> | ||
+ | </ul> | ||
+ | |||
+ | To see the design of this part go to the Design tab. | ||
+ | |||
+ | <h2> How to use it </h2> | ||
+ | |||
+ | The only requirement is to adapt the promotor sequence to the MoClo standard, adding the appropiate restriction sites at the ends. | ||
+ | |||
+ | <partinfo>BBa_K3122003 SequenceAndFeatures</partinfo> | ||
− | |||
− | |||
<partinfo>BBa_K3122003 parameters</partinfo> | <partinfo>BBa_K3122003 parameters</partinfo> | ||
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Latest revision as of 09:42, 22 November 2019
Constitutive promoter J23107 (TypeIIS adapted)
This part is the constitutive medium promoter, from Anderson family, was adapted to type IIS enzymes standard.
It has been used to express our Level 1 constructs related to membrane proteins. The reason behind this choice relays on its relative strength among other members of the family: it is a medium- strength regulator (0.36) which should not express too much membrane protein as to induce cell lysis.
Characterization
Characterization of this part has been conducted by building two transcriptional units (BBa_K3122004, BBa_ K3122005), and performing Lambda fague and Cell separation assays.
BBa_K3122004 transcriptional unit was assembled in pARK1 alpha plasmid including the following parts:
- BBa_K3122003: Promoter J23107 adapted to type IIS assembly
- BBa_K2656009: RBS B0030 adapted to type IIS assembly
- BBa_K3122002: coding sequence. This composite part is built with BBa_K3122000 (LamB protein) and BBa_K3122001 (6xHis tag) creating our AEGIS' lamB display system exposing 6xHis Tag.
- BBa_K2656026: Terminator B0015 adapted to type IIS assembly
BBa_K3122005 transcriptional unit was assembled in pARK1 alpha plasmid including the following parts:
- BBa_K3122003: Promoter J23107 adapted to type IIS assembly
- BBa_K2656009: RBS B0030 adapted to type IIS assembly
- BBa_K3122000: LamB coding sequence. This part is our AEGIS' lamB display system.
- BBa_K2656026: Terminator B0015 adapted to type IIS assembly
To see the design of this part go to the Design tab.
How to use it
The only requirement is to adapt the promotor sequence to the MoClo standard, adding the appropiate restriction sites at the ends.
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 11
Illegal NheI site found at 34 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]