Difference between revisions of "Part:BBa K3505013"
Line 5: Line 5:
 
Constitutive Anderson <bbpart>BBa_J23115</bbpart>with <i>Lac Operator</i>  and RBS <bbpart>BBa_B0034</bbpart>
 
Constitutive Anderson <bbpart>BBa_J23115</bbpart>with <i>Lac Operator</i>  and RBS <bbpart>BBa_B0034</bbpart>
 
This part is a constitutive promoter from Andesron family , J23115 , having downstream with a Lac Operator and RBS. The activity of this part is regulated by LacI  <bbpart>BBa_K3505003</bbpart>. LacI binds to Lac Operator and inhibits the expression of the desirable transcription unit.  
 
This part is a constitutive promoter from Andesron family , J23115 , having downstream with a Lac Operator and RBS. The activity of this part is regulated by LacI  <bbpart>BBa_K3505003</bbpart>. LacI binds to Lac Operator and inhibits the expression of the desirable transcription unit.  
−
[[File:T--Thessaly--anderson.png|700px|thumb|none|<i><b>Fig.1:</b>J23115 with LacO and RBS</i>]]
 
  
  
Line 20: Line 19:
 
===Verification of cloning===
 
===Verification of cloning===
  
−
[[File:T--Thessaly--LACO--digestion.png|700px|thumb|none|<i><b>Fig.1:</b>((U=Uncut , C= Cut)Restriction digestion of LacO (C3 & C4) with : EcoRV + HindIII (C3 & C4) , Expected bands : 2173bp ,Positive result : C3 + C4</i>]]
+
[[File:T--Thessaly--LACO--digestion.png|700px|thumb|none|<i><b>Fig.3:</b>((U=Uncut , C= Cut)Restriction digestion of LacO (C3 & C4) with : EcoRV + HindIII (C3 & C4) , Expected bands : 2173bp ,Positive result : C3 + C4</i>]]
  
  

Revision as of 16:15, 27 October 2020


AndersonJ23115:LacO:RBS GB compatible A1-B2

Constitutive Anderson BBa_J23115with Lac Operator and RBS BBa_B0034 This part is a constitutive promoter from Andesron family , J23115 , having downstream with a Lac Operator and RBS. The activity of this part is regulated by LacI BBa_K3505003. LacI binds to Lac Operator and inhibits the expression of the desirable transcription unit.



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 Golden Braid Grammar
Fig.2:The level 0 module :pupd2- AndersonJ23115:LacO:RBS I (illustration from SnapGene)

Verification of cloning

Fig.3:((U=Uncut , C= Cut)Restriction digestion of LacO (C3 & C4) with : EcoRV + HindIII (C3 & C4) , Expected bands : 2173bp ,Positive result : C3 + C4


Experimental Use and Experience

This part is used in BBa_K3505032

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]