Difference between revisions of "Part:BBa K3505014"
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===Verification of cloning=== | ===Verification of cloning=== | ||
− | [[File:T--Thessaly--TET0-digestion.png | | + | [[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=== |
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.
Verification of cloning
Experimental Use and Experience
This part is used in BBa_K3505034
Source
Synthesized by IDT.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 11
Illegal NheI site found at 34 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]