Difference between revisions of "Part:BBa K1648051"
 
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This construct composed of constitutive promoter, rbs, K1172501 and HoxKGZ flanking sequence. As the HoxKGZ flanking sequence was a hydrogenase flanking sequence amplified from Azotobacter vinelandii (A. vinelandii), the part would overexpress both porin and hydrogenase once transformed into A. vinelandii with homologous recombination. A BamHI site was included in the middle of the flanking sequence for linearizing the plasmid DNA before transformation. (After cutting with BamHI the 5' sequence in front of HoxKGZ would be at the beginning and the CDS of HoxKGZ would be at the end of the sequence so an insertion would occurred right in front of the CDS of HoxKGZ.)
 
This construct composed of constitutive promoter, rbs, K1172501 and HoxKGZ flanking sequence. As the HoxKGZ flanking sequence was a hydrogenase flanking sequence amplified from Azotobacter vinelandii (A. vinelandii), the part would overexpress both porin and hydrogenase once transformed into A. vinelandii with homologous recombination. A BamHI site was included in the middle of the flanking sequence for linearizing the plasmid DNA before transformation. (After cutting with BamHI the 5' sequence in front of HoxKGZ would be at the beginning and the CDS of HoxKGZ would be at the end of the sequence so an insertion would occurred right in front of the CDS of HoxKGZ.)
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https://static.igem.org/mediawiki/2015/b/b4/PfOprF_with_HoxKGZ.png
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Gel photo of double digested Biobrick with BamHI and EcoRI (takara 1kb ladder)
  
 
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Latest revision as of 19:17, 17 September 2015

PfOprF with HoxKGZ flanking sequences

This construct composed of constitutive promoter, rbs, K1172501 and HoxKGZ flanking sequence. As the HoxKGZ flanking sequence was a hydrogenase flanking sequence amplified from Azotobacter vinelandii (A. vinelandii), the part would overexpress both porin and hydrogenase once transformed into A. vinelandii with homologous recombination. A BamHI site was included in the middle of the flanking sequence for linearizing the plasmid DNA before transformation. (After cutting with BamHI the 5' sequence in front of HoxKGZ would be at the beginning and the CDS of HoxKGZ would be at the end of the sequence so an insertion would occurred right in front of the CDS of HoxKGZ.)


PfOprF_with_HoxKGZ.png Gel photo of double digested Biobrick with BamHI and EcoRI (takara 1kb ladder)

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 838
    Illegal BamHI site found at 1624
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1230
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 751