Regulatory

Part:BBa_K3505013

Designed by: Asteria Tsapadikou   Group: iGEM20_Thessaly   (2020-10-20)
Revision as of 16:15, 27 October 2020 by Asteriat (Talk | contribs)


AndersonJ23115:LacO:RBS GB compatible A1-B2

Constitutive Anderson BBa_J23115with Lac Operator and RBS BBa_B0034


Usage and Biology

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.



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.3:((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]
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Categories
Parameters
None