Difference between revisions of "Part:BBa K3505033"
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===Usage and Biology=== | ===Usage and Biology=== | ||
− | + | This parts consists of AndersonJ23115 promoter <bbpart>BBa_K3505012</bbpart> , TetR <bbpart>BBa_K3505005</bbpart> and Double Terminator <bbpart>BBa_K3505017</bbpart>. By this way ,as AndersonJ23115 is a strong constitutive promoter, TetR is constitutively expressed. TetR binds to tetracycline response element and inhibits the expression of a desirable transcription unit (Gossen and Bujard, 1993). | |
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===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 alpha1R vector <bbpart>BBa_K3505008</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 alpha1R vector <bbpart>BBa_K3505008</bbpart> and has overhangs compatible for GoldenBraid cloning. |
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+ | [[Image:T--Thessaly--ANDERSON-TETR-PHOT.png|900px|thumb|none|<I><b>Figure 1.</b> The level A module of Tet-Off system : AndersonJ23115:RBS-TetR-Double terminator </i>]] | ||
===Verification of Cloning=== | ===Verification of Cloning=== | ||
− | [[File:T--Thessaly--Anderson--TETR-LEVEL1.png|700px|thumb|none|<i><b>Fig. | + | [[File:T--Thessaly--Anderson--TETR-LEVEL1.png|700px|thumb|none|<i><b>Fig.2:</b>(U=Uncut C=Cut) Restriction digestion of T: AndersonJ23115-TetR-double terminator(C3-C4 ) with: HindIII (C3-C4) , Expected bands : 2847 + 836 bp ,Positive result : C3,C4</i>]] |
===Sequence and Features=== | ===Sequence and Features=== | ||
<partinfo>BBa_K3505033 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3505033 SequenceAndFeatures</partinfo> | ||
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+ | ===References=== | ||
+ | *Gossen, M. and Bujard, H., 1993. Anhydrotetracycline, a novel effector for tetracycline controlled gene expression systems in eukaryotic cells. Nucleic Acids Research, 21(18), pp.4411-4412. |
Latest revision as of 10:06, 14 September 2021
pAndersonJ23115:RBS:tetR-terminator
Usage and Biology
This parts consists of AndersonJ23115 promoter BBa_K3505012 , TetR BBa_K3505005 and Double Terminator BBa_K3505017. By this way ,as AndersonJ23115 is a strong constitutive promoter, TetR is constitutively expressed. TetR binds to tetracycline response element and inhibits the expression of a desirable transcription unit (Gossen and Bujard, 1993).
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 alpha1R vector BBa_K3505008 and has overhangs compatible for GoldenBraid cloning.
Verification of Cloning
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 76
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 11
Illegal NheI site found at 34 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 76
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 76
- 1000COMPATIBLE WITH RFC[1000]
References
- Gossen, M. and Bujard, H., 1993. Anhydrotetracycline, a novel effector for tetracycline controlled gene expression systems in eukaryotic cells. Nucleic Acids Research, 21(18), pp.4411-4412.