Translational_Unit

Part:BBa_K1465205:Design

Designed by: iGEM-Team Bielefeld 2014   Group: iGEM14_Bielefeld-CeBiTec   (2014-09-22)


Carbonic anhydrase (csoS3) of the carboxysome of Halothiobacillus neapolitanus


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 NgoMIV site found at 166
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

  • Amplification of the pSB1C3-Backbone (BBa_J04450) using the specific Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#pSB1C3_pre_can pSB1C3_pre_can] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#pSB1C3_suf_can pSB1C3_suf_can]:
pSB1C3_pre_can (46 bp)

5'-GTTCATGATCGTTACACTTTACTTAACTCTAGAAGCGGCCGCGAAT-3'

pSB1C3_suf_can (46 bp)

5'-TCGAAGAGGTTGCATCCGCATGATACTAGTAGCGGCCGCTGCAGTC-3'
  • Amplification of coding sequence of the carbonic anhydrase csoS3 (can) from the Plasmid [http://www.addgene.org/52065/ pHnCBS1D] containing also the natural interspace with the RBS using specific the Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#csoS3_can_fwd csoS3_can_fwd] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#csoS3_can_rev csoS3_can_rev]:
csoS3_can_fwd (50 bp)

5'-ATTCGCGGCCGCTTCTAGAGTTAAGTAAAGTGTAACGATCATGAACACCC-3'

csoS3_can_rev (41 bp)

5'-TGCAGCGGCCGCTACTAGTATCATGCGGATGCAACCTCTTC-3'
  • [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Gibson Gibson-Assembly] of the pSB1C3-Backbone and the coding sequence of carbonic anhydrase csoS3 (can) containing also the natural interspace region with RBS from Halothiobacillus neapolitanus.

Source

  • [http://www.ncbi.nlm.nih.gov/nucleotide/261835099?report=genbank&log$=nuclalign&blast_rank=1&RID=2T27D737014 Halothiobacillus neapolitanus c2 (GenBank: CP001801.1)]: Hneap 0919
  • Plasmid [http://www.addgene.org/52065/ pHnCBS1D] from Bonacci et al, 2011

References

  • Walter Bonacci, Poh K. Teng, Bruno Afonso, Henrike Niederholtmeyer, Patricia Grob, Pamela A. Silver

and David F. Savage (2011) Modularity of a carbon-fixing protein organelle [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3258634/pdf/pnas.201108557.pdf Proc Natl Acad Sci U S A. 109: 478–483.]