Difference between revisions of "Part:BBa K1955008:Design"

 
(→‎Design Notes)
 
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
Use the standard parts provided by iGEM, we choose BBa_J04500 as a promoter to induce production of protein. BBa_J04500 is a LacI inducible promoter with RBS, so it can be induced by IPTG to open its promoter and start transcription. Double restriction enzyme cutting(EcoRI and XbaI) of pSB1C3-HA to produce a sticky end in the anterior, then also double restriction enzyme cutting(EcoRI and SpeI) of BBa_J04500 to produce other sticky end which can be ligated with EcoRI and XbaI sticky end. Moreover, it will form a scar when XbaI ligate with SpeI, so we can simply compose the pSB1C3-J04500-HA.
+
Among the standard parts provided by iGEM, we chose BBa_J04500 as a promoter to induce production of protein. BBa_J04500 is a LacI inducible promoter with RBS, so it can be induced by IPTG to activate its promoter and start transcription. Double restriction enzyme cutting (EcoRI and XbaI) of pSB1C3-HA to produce a sticky end in the anterior, then also double restriction enzyme cutting (EcoRI and SpeI) of BBa_J04500 to produce other sticky end which can be ligated with EcoRI and XbaI sticky end. Moreover, it will form a scar when XbaI ligate with SpeI, so we can simply compose the pSB1C3-J04500-HA.
−
 
+
−
 
+
  
 
===Source===
 
===Source===

Latest revision as of 11:38, 19 October 2016


pSB1C3-LacI inducible Hemagglutinin(H1N1)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 232
    Illegal BamHI site found at 1938
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1868


Design Notes

Among the standard parts provided by iGEM, we chose BBa_J04500 as a promoter to induce production of protein. BBa_J04500 is a LacI inducible promoter with RBS, so it can be induced by IPTG to activate its promoter and start transcription. Double restriction enzyme cutting (EcoRI and XbaI) of pSB1C3-HA to produce a sticky end in the anterior, then also double restriction enzyme cutting (EcoRI and SpeI) of BBa_J04500 to produce other sticky end which can be ligated with EcoRI and XbaI sticky end. Moreover, it will form a scar when XbaI ligate with SpeI, so we can simply compose the pSB1C3-J04500-HA.

Source

LacI inducible promoter came from BBa_J04500; hemagglutinin came from BBa_K1955003.

References