Difference between revisions of "Part:BBa K3505036"
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<partinfo>BBa_K3505036 short</partinfo>
 
<partinfo>BBa_K3505036 short</partinfo>
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[[File:T--Thessaly--pflic.jpeg|600px|thumb|none|<i><b>Fig.1:</b> Activation of eCFP in absence of SCFAs,Based on NOT GATE</i>]]
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[[File:T--Thessaly--notgate.png|600px|thumb|none|<i><b>Fig.2:</b> The NOT GATE</i>]]
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===Usage and Biology===
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2 Trancription Units
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*LacI <bbpart>BBa_K3505003</bbpart>regulated by inducible promoter FliC <bbpart>BBa_K2924016</bbpart>which is activated from SFCAs and more specifically from Butyrate
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*eCFP <bbpart>BBa_K3505019</bbpart>under control of lac regulated Anderson<bbpart>BBa_K3505013</bbpart>.
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<b>This way we take signal in the Absence of SCFAs</b>
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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 present in <bbpart>BBa_K3505010</bbpart> and has overhangs compatible for Golden Braid cloning.
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===Verification of cloning===
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[[File:T--Thessaly--pflicfinal.png|600px|thumb|none|<i><b>Fig.3:</b>U2 C2  pFliC:RBS-LacI-Terminator-pAndersonJ23115:LacO:RBS-ECFP-terminator  EcoRV, HindIII. Expected bands 3734, 1699.</i>]]
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===Experimental Use and Experience===
  
  

Revision as of 17:18, 24 October 2020


pFliC:RBS-LacI-Terminator- pAndersonJ23115:LacO:RBS-ECFP-terminator

Fig.1: Activation of eCFP in absence of SCFAs,Based on NOT GATE
Fig.2: The NOT GATE


Usage and Biology

2 Trancription Units

This way we take signal in the Absence of SCFAs


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 present in BBa_K3505010 and has overhangs compatible for Golden Braid cloning.



Verification of cloning

Fig.3:U2 C2 pFliC:RBS-LacI-Terminator-pAndersonJ23115:LacO:RBS-ECFP-terminator EcoRV, HindIII. Expected bands 3734, 1699.

Experimental Use and Experience

Sequnce and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1321
    Illegal NheI site found at 1344
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]