Part:BBa_K1465222:Design

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