Difference between revisions of "Part:BBa K3505018"
 
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<partinfo>BBa_K3505018 short</partinfo>
 
<partinfo>BBa_K3505018 short</partinfo>
  
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Superfolder GFP derived from <i>Aequorea victoria</i> with weak dimer oligomerization
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Level 0 sfGFP
  
−
[[File:T--Thessaly--GB-AATG-GCTT.jpeg|600px|thumb|none|<i><b>Fig.1:</b>The overhangs of this part in the Golden Braid Grammar</i>]]
+
[[File:T--Thessaly--sf.png|700px|thumb|none|<i><b>Fig.1:</b>sfGFP</i>]]
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 +
 
 +
 
 +
[[File:T--Thessaly--sfpiato.jpeg|700px|thumb|none|<i><b>Fig.2:</b>sfGFP uder control of pTRC</i>]]
  
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[[File:T--Thessaly--sfpiato.png|600px|thumb|none|<i><b>Fig.2:</b>sfGFP uder control of pTRC</i>]]
 
  
 
===Usage and Biology===
 
===Usage and Biology===
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===Design Notes===
 
===Design Notes===
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The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is presents in pUPD2 and has overhangs compatible for Golden Braid cloning.  
+
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 <bbpart>BBa_K3505007</bbpart>and has overhangs compatible for GoldenBraid cloning.  
 
The CDS has position B3-B5.
 
The CDS has position B3-B5.
  
−
 
+
[[File:T--Thessaly--GB-AATG-GCTT.jpeg|700px|thumb|none|<i><b>Fig.3:</b>The overhangs of this part in the Golden Braid Grammar</i>]]
  
 
===Verification of cloning===
 
===Verification of cloning===
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[[File:T--Thessaly--sf0.png|600px|thumb|none|<i><b>Fig.3:</b>U4 C4 Level0 sfGFP Digested with EcoRI, PstI Expected bands 2029, 794</i>]]
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[[File:T--Thessaly--sf0.png|600px|thumb|none|<i><b>Fig.4:</b>U4 C4 Level0 sfGFP Digested with EcoRI, PstI Expected bands 2029, 794</i>]]
  
 
===Experimental Use and Experience===
 
===Experimental Use and Experience===
  
 
This part is used in <bbpart>BBa_K3505030</bbpart>
 
This part is used in <bbpart>BBa_K3505030</bbpart>
 +
===Source===
 +
Christos Batianis [1]
  
 
===Sequence and Features===
 
===Sequence and Features===
 
<partinfo>BBa_K3505018 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3505018 SequenceAndFeatures</partinfo>
 +
 +
===References===
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*[1]Damalas, S., Batianis, C., Martin‐Pascual, M., Lorenzo, V. and Martins dos Santos, 2020. SEVA 3.1: enabling interoperability of DNA assembly among the SEVA, BioBricks and Type IIS restriction enzyme standards. <i>Microbial Biotechnology</i>, 13(6), pp.1793-1806.

Latest revision as of 00:10, 28 October 2020


sfGFP GB compatible with B3-B5

Superfolder GFP derived from Aequorea victoria with weak dimer oligomerization 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

Source

Christos Batianis [1]

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]

References

  • [1]Damalas, S., Batianis, C., Martin‐Pascual, M., Lorenzo, V. and Martins dos Santos, 2020. SEVA 3.1: enabling interoperability of DNA assembly among the SEVA, BioBricks and Type IIS restriction enzyme standards. Microbial Biotechnology, 13(6), pp.1793-1806.