Reporter

Part:BBa_K3505018

Designed by: Venetios Michelioudakis   Group: iGEM20_Thessaly   (2020-10-20)
Revision as of 20:46, 27 October 2020 by Asteriat (Talk | contribs) (→‎Design Notes)


sfGFP GB compatible with B3-B5


Level 0 sfGFP

Fig.1:sfGFP


Fig.2:sfGFP uder control of pTRC


Usage and Biology

A reporter gene fluorescent protein

  • Excitation at 485 nm
  • Emission at 510 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 GoldenBraid cloning. The CDS has position B3-B5.

Fig.3:The overhangs of this part in the Golden Braid Grammar

Verification of cloning

Fig.4:U4 C4 Level0 sfGFP Digested with EcoRI, PstI Expected bands 2029, 794

Experimental Use and Experience

This part is used in BBa_K3505030

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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 149
  • 1000
    COMPATIBLE WITH RFC[1000]
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Categories
Parameters
None