Difference between revisions of "Part:BBa K3505034"
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===Usage and Biology=== | ===Usage and Biology=== | ||
− | This parts consists of AndersonJ23115 promoter <bbpart>BBa_K3505012</bbpart> , LacI <bbpart>BBa_K3505003</bbpart> and Double Terminator <bbpart>BBa_K3505017</bbpart>. By this way ,as AndersonJ23115 is a strong constitutive promoter, LacI is constitutively expressed. LacI binds to lac operator and inhibits the expression of a desirable transcription unit | + | This parts consists of AndersonJ23115 promoter <bbpart>BBa_K3505012</bbpart> , LacI <bbpart>BBa_K3505003</bbpart> and Double Terminator <bbpart>BBa_K3505017</bbpart>. By this way ,as AndersonJ23115 is a strong constitutive promoter, LacI is constitutively expressed. LacI binds to lac operator and inhibits the expression of a desirable transcription unit. |
===Design Notes=== | ===Design Notes=== | ||
− | The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in alpha vector <bbpart>BBa_K3505009</bbpart> and has overhangs compatible for | + | The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in alpha vector <bbpart>BBa_K3505009</bbpart> and has overhangs compatible for GoldenBraid cloning. |
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+ | [[Image:T--Thessaly--teO-LacI-PHOT.png|900px|thumb|none|<I><b>Figure 1.</b> The level A module of Tet-Off system : AndersonJ23115:TetO:RBS-LacI-Double terminator </i>]] | ||
===Verification of Cloning=== | ===Verification of Cloning=== | ||
− | [[File:T--Thessaly--tetO-LacI-digestionpng.png|700px|thumb|none|<i><b>Fig. | + | [[File:T--Thessaly--tetO-LacI-digestionpng.png|700px|thumb|none|<i><b>Fig.2:</b>(U=Uncut C=Cut) Restriction digestion of TL: AndersonJ23115:Tet0-LacI-double terminator(C5-C8 ) with: HindIII + NheI (C5-C18) , Expected bands : 2573+891+427,Positive result : C5,C6</i>]] |
===Sequence and Features=== | ===Sequence and Features=== | ||
<partinfo>BBa_K3505034 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3505034 SequenceAndFeatures</partinfo> |
Latest revision as of 00:41, 28 October 2020
pAndersonJ23115:tetO:RBS-lacI-terminator
Usage and Biology
This parts consists of AndersonJ23115 promoter BBa_K3505012 , LacI BBa_K3505003 and Double Terminator BBa_K3505017. By this way ,as AndersonJ23115 is a strong constitutive promoter, LacI is constitutively expressed. LacI binds to lac operator and inhibits the expression of a desirable transcription unit.
Design Notes
The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in alpha vector BBa_K3505009 and has overhangs compatible for GoldenBraid cloning.
Verification of Cloning
Sequence and Features
Assembly Compatibility:
- 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]