Difference between revisions of "Part:BBa K1172501:Design"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K1172501 short</partinfo> | <partinfo>BBa_K1172501 short</partinfo> | ||
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===Design Notes=== | ===Design Notes=== | ||
− | + | '''Amplification with Gibson-primer:''' | |
+ | *Amplification of the outer membrane protein (''oprF''):<br> | ||
+ | |||
+ | '''Forward primer oprF (49 bp)'''<br> | ||
+ | 5'-GAATTCGCGGCCGCTTCTAGATGAAACTGAAAAACACCTTGGGCTTTGC-3'<br> | ||
+ | '''Reverse primer oprF (51 bp)'''<br> | ||
+ | 5'-CTGCAGCGGCCGCTACTAGTATTACTTAGCTTGGGCTTCAACCTGCGCTTC-3'<br> | ||
+ | |||
+ | *Amplification of the pSB1C3-Backbone:<br> | ||
+ | |||
+ | '''Forward primer pSB1C3 oprF (43 bp)'''<br> | ||
+ | 5'-TTGAAGCCCAAGCTAAGTAATACTAGTAGCGGCCGCTGCAGTC-3'<br> | ||
+ | '''Reverse primer pSB1C3 oprF (43 bp)'''<br> | ||
+ | 5'-AAGGTGTTTTTCAGTTTCATCTAGAAGCGGCCGCGAATTCCAG-3'<br> | ||
+ | |||
+ | |||
+ | |||
+ | '''Ligation via [http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/ProtocolsPrograms#Gibson_assembly Gibson-Assembly].''' | ||
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===Source=== | ===Source=== | ||
− | + | ''Pseudomonas fluorescens'' Migula 1895 genomic DNA ([http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/ProtocolsPrograms#Whole_Genome_Isolation Whole Genome Isolation]) | |
+ | '''DSMZ No.: 7355''' | ||
===References=== | ===References=== |
Latest revision as of 15:37, 22 October 2013
Outer membrane porin OprF from Pseudomonas fluorescens
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 764
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1156
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 677
Design Notes
Amplification with Gibson-primer:
- Amplification of the outer membrane protein (oprF):
Forward primer oprF (49 bp)
5'-GAATTCGCGGCCGCTTCTAGATGAAACTGAAAAACACCTTGGGCTTTGC-3'
Reverse primer oprF (51 bp)
5'-CTGCAGCGGCCGCTACTAGTATTACTTAGCTTGGGCTTCAACCTGCGCTTC-3'
- Amplification of the pSB1C3-Backbone:
Forward primer pSB1C3 oprF (43 bp)
5'-TTGAAGCCCAAGCTAAGTAATACTAGTAGCGGCCGCTGCAGTC-3'
Reverse primer pSB1C3 oprF (43 bp)
5'-AAGGTGTTTTTCAGTTTCATCTAGAAGCGGCCGCGAATTCCAG-3'
Ligation via [http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/ProtocolsPrograms#Gibson_assembly Gibson-Assembly].
Source
Pseudomonas fluorescens Migula 1895 genomic DNA ([http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/ProtocolsPrograms#Whole_Genome_Isolation Whole Genome Isolation]) DSMZ No.: 7355