Difference between revisions of "Part:BBa K1256004"
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[[File: Mingdao_pSB1C3-Plac-SS-Cecropin-MprF.png|400px]] | [[File: Mingdao_pSB1C3-Plac-SS-Cecropin-MprF.png|400px]] | ||
| − | MprF of Bacillus subtilis (DB2)added a RBS (BBa_B0034)was amplified by PCR and cloned into the vector of [https://parts.igem.org/Part:BBa_K1256003 pSB1C3-Plac-SS-NB (Part:BBa_K1256003)] using NheI and BamHI sites. Next, the cds of cecropin was synthesized on the primers and subjeted to Gibson Assembly to make this plasmid. The amino acid sequences of cecropin was obtained from Sigma-Aldrich, and the nucleotide sequence was optimized for engineering Escherichia coli using the [http://sg.idtdna.com/CodonOpt codon optimization] tool provided by Integrated DNA Technologies (IDT). | + | MprF of Bacillus subtilis (DB2)added a RBS (BBa_B0034)was amplified by PCR and cloned into the vector of [https://parts.igem.org/Part:BBa_K1256003 pSB1C3-Plac-SS-NB (Part:BBa_K1256003)] using NheI and BamHI sites. Next, the cds of cecropin was synthesized on the primers and subjeted to Gibson Assembly to make this plasmid. The [http://www.sigmaaldrich.com/catalog/product/sigma/c6830?lang=en®ion=TW amino acid sequences] of cecropin was obtained from Sigma-Aldrich, and the nucleotide sequence was optimized for engineering Escherichia coli using the [http://sg.idtdna.com/CodonOpt codon optimization] tool provided by Integrated DNA Technologies (IDT). |
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Latest revision as of 02:01, 10 June 2014
pSB1C3-Plac-SS-Cecropin-MprF
MprF of Bacillus subtilis (DB2)added a RBS (BBa_B0034)was amplified by PCR and cloned into the vector of pSB1C3-Plac-SS-NB (Part:BBa_K1256003) using NheI and BamHI sites. Next, the cds of cecropin was synthesized on the primers and subjeted to Gibson Assembly to make this plasmid. The [http://www.sigmaaldrich.com/catalog/product/sigma/c6830?lang=en®ion=TW amino acid sequences] of cecropin was obtained from Sigma-Aldrich, and the nucleotide sequence was optimized for engineering Escherichia coli using the [http://sg.idtdna.com/CodonOpt codon optimization] tool provided by Integrated DNA Technologies (IDT).
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 398
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2924
Illegal BamHI site found at 2993 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 371
Illegal NgoMIV site found at 1319
Illegal NgoMIV site found at 2375 - 1000COMPATIBLE WITH RFC[1000]