Difference between revisions of "Part:BBa K3505044"
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<partinfo>BBa_K3505044 short</partinfo> | <partinfo>BBa_K3505044 short</partinfo> | ||
+ | eCFP <bbpart>BBa_K3505020</bbpart>uder control of a constitutive promoter (andersonJ23115 with a tet operator) <bbpart>BBa_K2924014</bbpart>. | ||
− | + | [[File:T--Thessaly--TET0-ECFP-PHOT.png|600px|thumb|none|<i><b>Fig.1:</b>The Level A construct AndersonJ23115:TetO:rbs-eCFP-Terminator</i>]] | |
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===Usage and Biology=== | ===Usage and Biology=== | ||
+ | This Trancriscription Unit (TU) is continuesly activated exressing the eCFP protein as a reporter. The Lac operator that is downsteam the anderson exists for the lac regulated inhibition. | ||
− | + | ===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 a1R <bbpart>BBa_K3505008</bbpart> and has overhangs compatible for Golden Braid cloning. | |
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+ | ===Verification of cloning=== | ||
+ | [[File:T--Thessaly--TET0-ECFP-digestion.png|600px|thumb|none|<i><b>Fig.2:</b>(U=Uncut C=Cut) (U=Uncut , C= Cut) Restriction digestion of TE: AndersonJ23115:Tet0-EGFP-double terminator(C1-C 4) with : EcoRV(C1-C2) , Expected bands : 2847+ 954 bp ,Positive result: C1,C2</i>]] | ||
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− | === | + | ===Sequence and Features=== |
− | <partinfo>BBa_K3505044 | + | <partinfo>BBa_K3505044 SequenceAndFeatures</partinfo> |
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Revision as of 00:26, 27 October 2020
pAndersonJ23115:TetO:RBS-eCFP -terminator
eCFP BBa_K3505020uder control of a constitutive promoter (andersonJ23115 with a tet operator) BBa_K2924014.
Usage and Biology
This Trancriscription Unit (TU) is continuesly activated exressing the eCFP protein as a reporter. The Lac operator that is downsteam the anderson exists for the lac regulated inhibition.
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 a1R BBa_K3505008 and has overhangs compatible for Golden Braid cloning.
Verification of cloning
Sequence and Features
Assembly Compatibility:
- 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]