Difference between revisions of "Part:BBa K3505037"

 
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===Usage and Biology===
 
===Usage and Biology===
  
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<span class='h3bb'>Sequence and Features</span>
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===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 omega1 vector <bbpart>BBa_K3505010</bbpart> and has overhangs compatible for Golden Braid cloning.
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[[Image:T--Thessaly--omega1-tetoff-PHOT.png|900px|thumb|none|<I><b>Figure 2.</b> The level B module of Tet-Off system : a1R:AndersonJ23115:RBS-TetR-Double terminator-a2:AndersonJ23115:TetO:RBS- LacI-Double terminator </i>]]
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===Verification of Cloning===
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[[File:T--Thessaly--omga1-tetoff-digestion.png|700px|thumb|none|<i><b>Fig.1:</b>: (U=Uncut , C= Cut) Restriction digestion of omega1R-TetR-LacI (C1-C 4) with : �EcoRV +EcoRI(C1-C4) , Expected bands : 4823 + 465bp ,Positive result: C1,C2,C3,C4</i>]]
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===Sequence and Features===
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<partinfo>BBa_K3505034 SequenceAndFeatures</partinfo>
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===Sequence and Features</span>===
 
<partinfo>BBa_K3505037 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3505037 SequenceAndFeatures</partinfo>
  

Revision as of 14:33, 26 October 2020


pAndersonJ23115:RBS-TetR-terminator- pAndersonJ23115:tetO:RBS-LacI-terminator

long


Usage and Biology

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 omega1 vector BBa_K3505010 and has overhangs compatible for Golden Braid cloning.

Figure 2. The level B module of Tet-Off system : a1R:AndersonJ23115:RBS-TetR-Double terminator-a2:AndersonJ23115:TetO:RBS- LacI-Double terminator

Verification of Cloning

Fig.1:: (U=Uncut , C= Cut) Restriction digestion of omega1R-TetR-LacI (C1-C 4) with : �EcoRV +EcoRI(C1-C4) , Expected bands : 4823 + 465bp ,Positive result: C1,C2,C3,C4

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]


Sequence and Features</span>


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 76
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 11
    Illegal NheI site found at 34
    Illegal NheI site found at 838
    Illegal NheI site found at 861
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 76
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 76
  • 1000
    COMPATIBLE WITH RFC[1000]