Difference between revisions of "Part:BBa K3505014"
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===Usage and Biology===
 
===Usage and Biology===
 
This part is a constitutive promoter from Andesron family , J23115 , having downstream with a Τet Operator and RBS. The activity of this part is regulated by TetR  <bbpart>BBa_K3505003</bbpart>. TetR binds to TRE -tetracycline response element, and inhibits the expression of the desirable transcription unit.  
 
This part is a constitutive promoter from Andesron family , J23115 , having downstream with a Τet Operator and RBS. The activity of this part is regulated by TetR  <bbpart>BBa_K3505003</bbpart>. TetR binds to TRE -tetracycline response element, and inhibits the expression of the desirable transcription unit.  
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This part can be used in a Tet-off system, expression of Tet-regulated genes can be inhibited by tetracycline and its chemical derivatives , such as anhydrotetracycline aTc-a strong chemical repressor , that binds to TRE and the desirable transcription unit is blocked.
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+
−
 
+
−
 
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+
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+
  
 
===Design Notes===
 
===Design Notes===
 
The sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in pUPD2 <bbpart>BBa_K3505007</bbpart> and has overhangs compatible for Golden Braid cloning.  
 
The sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in pUPD2 <bbpart>BBa_K3505007</bbpart> and has overhangs compatible for Golden Braid cloning.  
 
This has position A1-B3.
 
This has position A1-B3.
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[[File:T--Thessaly--GB-GGAG-AATG.jpeg|700px|thumb|none|<i><b>Fig.1:</b>The overhangs of this part in the Golden Braid Grammar</i>]]
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[[File:T--Thessaly--GB-GGAG-AATG.jpeg|700px|thumb|none|<i><b>Fig.1:</b>The overhangs of this part in the GoldenBraid Grammar</i>]]
  
 
[[Image:T--Thessaly--andersontetO-snap.png|900px|thumb|none|<I><b>Figure 2.</b> The level 0 module : pupd2- AndersonJ23115:TetO:RBS (illustration from SnapGene)</i>]]
 
[[Image:T--Thessaly--andersontetO-snap.png|900px|thumb|none|<I><b>Figure 2.</b> The level 0 module : pupd2- AndersonJ23115:TetO:RBS (illustration from SnapGene)</i>]]
  
 
===Verification of cloning===
 
===Verification of cloning===
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[[File:T--Thessaly--TET0-digestion.png |700px|thumb|none|<i><b>Fig.1:</b>Figure 7: (U=Uncut , C= Cut) Restriction digestion of TetO (C1-C4 ) with :Not-HF(C1-C4) , Expected bands : 2046 + 129 bp, Positive result: C1,C2,C4</i>]]
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[[File:T--Thessaly--TET0-digestion.png |500px|thumb|none|<i><b>Fig.3:</b> (U=Uncut , C= Cut) Restriction digestion of TetO (C1-C4 ) with :Not-HF(C1-C4) , Expected bands : 2046 + 129 bp, Positive result: C1,C2,C4</i>]]
  
 
===Experimental Use and Experience===
 
===Experimental Use and Experience===
  
 
This part is used in <bbpart>BBa_K3505034</bbpart>
 
This part is used in <bbpart>BBa_K3505034</bbpart>
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 +
===Source===
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Synthesized by IDT.
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===Sequence and Features===
 
===Sequence and Features===
 
<partinfo>BBa_K3505014 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3505014 SequenceAndFeatures</partinfo>

Latest revision as of 00:40, 28 October 2020


AndersonJ23115:TetO:RBS GB compatible with A1-B2

Constitutive Anderson BBa_J23115with Tet Operator and RBS BBa_B0034

Usage and Biology

This part is a constitutive promoter from Andesron family , J23115 , having downstream with a Τet Operator and RBS. The activity of this part is regulated by TetR BBa_K3505003. TetR binds to TRE -tetracycline response element, and inhibits the expression of the desirable transcription unit. This part can be used in a Tet-off system, expression of Tet-regulated genes can be inhibited by tetracycline and its chemical derivatives , such as anhydrotetracycline aTc-a strong chemical repressor , that binds to TRE and the desirable transcription unit is blocked.

Design Notes

The sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in pUPD2 BBa_K3505007 and has overhangs compatible for Golden Braid cloning. This has position A1-B3.

Fig.1:The overhangs of this part in the GoldenBraid Grammar
Figure 2. The level 0 module : pupd2- AndersonJ23115:TetO:RBS (illustration from SnapGene)

Verification of cloning

Fig.3: (U=Uncut , C= Cut) Restriction digestion of TetO (C1-C4 ) with :Not-HF(C1-C4) , Expected bands : 2046 + 129 bp, Positive result: C1,C2,C4

Experimental Use and Experience

This part is used in BBa_K3505034

Source

Synthesized by IDT.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 11
    Illegal NheI site found at 34
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]