Difference between revisions of "Part:BBa K1465222:Design"

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===Design Notes===
 
===Design Notes===
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*Amplification of the pSB1C3-Backbone containing the shell-proteins (<bbpart>BBa_K1465206</bbpart>) using the specific Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw-csoS1A-GFP fw-csoS1A-GFP] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv-csoS1B-GFP rv-csoS1B-GFP]:<br>
  
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'''fw-csoS1A-GFP (45 bp)'''<br>
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5'-CTGCCTAAGGCGCCACAAGCCATGCGTAAAGGAGAAGAACTTTTC-3'<br>
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'''rv-csoS1B-GFP (55 bp)'''<br>
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5'-GTGAGCCGTCTTAGGAATATCTGACTTATTATTTGTATAGTTCATCCATGCCATG-3'<br>
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*Amplification of coding sequence of the reporter GFP <bbpart>BBa_E0040</bbpart> using specific the Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw-GFP-csoS1B fw-GFP-csoS1B] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv-GFP-csoS1A rv-GFP-csoS1A]:<br>
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'''fw-GFP-csoS1B (48 bp)'''<br>
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5'-CATGGATGAACTATACAAATAATAAGTCAGATATTCCTAAGACGGCTC-3'<br>
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'''rv-GFP-csoS1A (45 bp)'''<br>
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5'-GAAAAGTTCTTCTCCTTTACGCATGGCTTGTGGCGCCTTAGGCAG-3'<br>
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*[http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Gibson Gibson-Assembly] of the pSB1C3-Backbone containing the shell-proteins of the carboxysome from ''Halothiobacillus neapolitanus'' and the coding sequence of GFP fused to shell-protein ''csoS1A''.<br>
  
 
===Source===
 
===Source===

Latest revision as of 01:30, 8 October 2014


Carboxysome (shell-proteins (csoS14)) with translational GFP fusion (under T7 and without csoS2)


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 704
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1853


Design Notes

  • Amplification of the pSB1C3-Backbone containing the shell-proteins (BBa_K1465206) using the specific Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw-csoS1A-GFP fw-csoS1A-GFP] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv-csoS1B-GFP rv-csoS1B-GFP]:
fw-csoS1A-GFP (45 bp)

5'-CTGCCTAAGGCGCCACAAGCCATGCGTAAAGGAGAAGAACTTTTC-3'

rv-csoS1B-GFP (55 bp)

5'-GTGAGCCGTCTTAGGAATATCTGACTTATTATTTGTATAGTTCATCCATGCCATG-3'
  • Amplification of coding sequence of the reporter GFP BBa_E0040 using specific the Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw-GFP-csoS1B fw-GFP-csoS1B] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv-GFP-csoS1A rv-GFP-csoS1A]:
fw-GFP-csoS1B (48 bp)

5'-CATGGATGAACTATACAAATAATAAGTCAGATATTCCTAAGACGGCTC-3'

rv-GFP-csoS1A (45 bp)

5'-GAAAAGTTCTTCTCCTTTACGCATGGCTTGTGGCGCCTTAGGCAG-3'
  • [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Gibson Gibson-Assembly] of the pSB1C3-Backbone containing the shell-proteins of the carboxysome from Halothiobacillus neapolitanus and the coding sequence of GFP fused to shell-protein csoS1A.

Source

BBa_K1465206: shell proteins of the carboxysome from Halothiobacillus neapolitanus.
BBa_E0040: green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP

References