Difference between revisions of "Part:BBa K3505013"

Revision as of 16:11, 27 October 2020

AndersonJ23115:LacO:RBS GB compatible A1-B2

Constitutive Anderson BBa_J23115with Lac Operator and RBS BBa_B0034 This part is a constitutive promoter from Andesron family , J23115 , having downstream with a Lac Operator and RBS. The activity of this part is regulated by LacI BBa_K3505003. LacI binds to Lac Operator and inhibits the expression of the desirable transcription unit.

File:T--Thessaly--anderson.png

Fig.1:J23115 with LacO and RBS

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.

Fig.1:The overhangs of this part in the Golden Braid Grammar

Fig.2:The level 0 module :pupd2- AndersonJ23115:LacO:RBS I (illustration from SnapGene)

Verification of cloning

Fig.1:((U=Uncut , C= Cut)Restriction digestion of LacO (C3 & C4) with : EcoRV + HindIII (C3 & C4) , Expected bands : 2173bp ,Positive result : C3 + C4

Experimental Use and Experience

This part is used in BBa_K3505032

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]

@@ Line 4: / Line 4: @@
 Constitutive Anderson <bbpart>BBa_J23115</bbpart>with <i>Lac Operator</i>   and RBS <bbpart>BBa_B0034</bbpart>
+This part is a constitutive promoter from Andesron family , J23115 , having downstream with a Lac Operator and RBS. The activity of this part is regulated by LacI  <bbpart>BBa_K3505003</bbpart>. LacI binds to Lac Operator and inhibits the expression of the desirable transcription unit.
 [[File:T--Thessaly--anderson.png|700px|thumb|none|<i><b>Fig.1:</b>J23115 with LacO and RBS</i>]]