Composite

Part:BBa_K3505044

Designed by: Asteria Tsapadikou   Group: iGEM20_Thessaly   (2020-10-26)
Revision as of 20:33, 27 October 2020 by Asteriat (Talk | contribs)


pAndersonJ23115:TetO:RBS-eCFP -terminator

eCFP BBa_K3505020uder control of a constitutive promoter (andersonJ23115 with a tet operator) BBa_K3505014.

Fig.1:The Level A construct AndersonJ23115:TetO:rbs-eCFP-Terminator

Usage and Biology

This Trancriscription Unit (TU) is continuesly activated exressing the eCFP protein as a reporter. The Tet operator that is downsteam the anderson exists for the TetR 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 GoldenBraid cloning.

Verification of cloning

Fig.2:(U=Uncut C=Cut) (U=Uncut , C= Cut) Restriction digestion of TE: AndersonJ23115:Tet0-ECFP-double terminator(C1-C 4) with : EcoRV(C1-C2) , Expected bands : 2847+ 954 bp ,Positive result: C1,C2

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