Regulatory

Part:BBa_K3505012

Designed by: Asteria Tsapadikou   Group: iGEM20_Thessaly   (2020-10-20)


AndersonJ23115:RBS GB compatible with A1-B2


Constitutive Anderson BBa_J23115with RBS BBa_B0034

AndersonJ23115 is a strong constitutive promoter that can be used to tune the expression level of constitutively expressed parts.

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 GoldenBraid cloning. This has position A1-B3.

Fig.1:The overhangs of this part in the GoldenBraid Grammar
Fig.2:J23115 with RBS

Verification of cloning

Fig.3:((U=Uncut , C= Cut) Restriction digestion of AndersonJ23115 with : EcoRI (C1a & C2a),Expected bands : 2156 bp, XhoI (C1b & C2b),Expected bands : 1264 + 892 bp Positive result: The set of C1a and C1b (same sample)

Experimental Use and Experience

This part is used in BBa_K3505033 BBa_K3505035

Source

Synthesized by IDT.


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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Parameters
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