Difference between revisions of "Part:BBa K1465207:Design"

Revision as of 23:36, 1 October 2014

Pore-forming protein (csoS1D) of the carboxysome of Halothiobacillus neapolitanus

Design Notes

• Amplification of the pSB1C3-Backbone using specific the Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#pSB1C3_pre_csoS1D pSB1C3_pre_csoS1D] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#pSB1C3_suf_csoS1D pSB1C3_suf_csoS1D]:
  

pSB1C3_pre_csoS1D (43 bp)

5'-CGCATCTTCCCTACTAGACACTCTAGAAGCGGCCGCGAATTCC-3'

pSB1C3_suf_csoS1D (43 bp)

5'-GTCGCGCTGAAGGGTTCTAATACTAGTAGCGGCCGCTGCAGTC-3'

• Amplification of the csoS1D containing also the natural interspace with the RBS using specific the Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#csoS1D_fwd csoS1D_fwd] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#csoS1D_rev csoS1D_rev]:
  

csoS1D_fwd (40 bp)

5'-ATTCGCGGCCGCTTCTAGAGTGTCTAGTAGGGAAGATGCG-3'

csoS1D_rev (41 bp)

5'-TGCAGCGGCCGCTACTAGTATTAGAACCCTTCAGCGCGACG-3'

• [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Gibson Gibson-Assembly] of the pSB1C3-Backbone and the coding sequence of csoS1D 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 0903

• [http://www.addgene.org/52065/ Plasmid] 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.]