Reporter

Part:BBa_K3505019

Designed by: Venetios Michelioudakis   Group: iGEM20_Thessaly   (2020-10-20)
Revision as of 15:05, 27 October 2020 by Venetios (Talk | contribs) (→‎Verification of cloning)


eGFP GB compatible with B3-B5

Level 0 eGFP

Fig.1:eGFP



Usage and Biology

  • Derived from jellyfish Aequeora victoria
  • A reporter gene fluorescent protein
  • Excitation at 488 nm
  • Emission at 515 nm

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 pUPD2 BBa_K3505007and has overhangs compatible for Golden Braid cloning. The CDS has position B3-B5.

Fig.2:The overhangs of this part in the Golden Braid Grammar

Verification of cloning

Fig.3::(U=Uncut C=Cut) Restriction digestion of EGFP (C1-C4 ) with : PvuII, Expected bands : 2826 bp , Positive Result : C4

Experimental Use and Experience

This part is used in BBa_K3505029

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 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