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

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===Design Notes===
 
===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]:<br>
+
*Amplification of the pSB1C3-Backbone (<bbpart>BBa_J04450</bbpart>) using the specific 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]:<br>
  
 
  '''pSB1C3_pre_csoS1D (43 bp)'''<br>
 
  '''pSB1C3_pre_csoS1D (43 bp)'''<br>
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  5'-GTCGCGCTGAAGGGTTCTAATACTAGTAGCGGCCGCTGCAGTC-3'<br>
 
  5'-GTCGCGCTGAAGGGTTCTAATACTAGTAGCGGCCGCTGCAGTC-3'<br>
  
−
*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]:<br>
+
*Amplification of the csoS1D 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#csoS1D_fwd csoS1D_fwd] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#csoS1D_rev csoS1D_rev]:<br>
  
 
  '''csoS1D_fwd (40 bp)'''<br>
 
  '''csoS1D_fwd (40 bp)'''<br>
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* [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.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
+
* Plasmid [http://www.addgene.org/52065/ pHnCBS1D] from Bonacci ''et al'', 2011
  
 
===References===
 
===References===

Latest revision as of 19:20, 3 October 2014


Pore-forming protein (csoS1D) 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
    COMPATIBLE WITH RFC[25]
  • 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_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 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#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 pore-froming protein 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
  • 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.]