Difference between revisions of "Part:BBa K3505034"

(→‎Design Notes)
 
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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.
 
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 2.</b> The level A module of Tet-Off system : AndersonJ23115:TetO:RBS-LacI-Double terminator </i>]]
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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===
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[[File:T--Thessaly--tetO-LacI-digestionpng.png|700px|thumb|none|<i><b>Fig.1:</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>]]
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[[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.

Figure 1. The level A module of Tet-Off system : AndersonJ23115:TetO:RBS-LacI-Double terminator

Verification of Cloning

Fig.2:(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

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]