Difference between revisions of "Part:BBa K1172501:Design"

(→‎Source)
(→‎Source)
 
(One intermediate revision by the same user not shown)
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1172501 short</partinfo>
 
<partinfo>BBa_K1172501 short</partinfo>
Line 7: Line 6:
  
 
===Design Notes===
 
===Design Notes===
−
Primeroverlap for Gibson-Assembly
+
'''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].'''
  
  
Line 13: Line 29:
 
===Source===
 
===Source===
  
−
''Pseudomonas fluorescens'' Migula 1895 genomic DNA  
+
''Pseudomonas fluorescens'' Migula 1895 genomic DNA ([http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/ProtocolsPrograms#Whole_Genome_Isolation Whole Genome Isolation])
 
'''DSMZ No.: 7355'''
 
'''DSMZ No.: 7355'''
  
 
===References===
 
===References===

Latest revision as of 15:37, 22 October 2013

Outer membrane porin OprF from Pseudomonas fluorescens


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 764
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1156
  • 1000
    INCOMPATIBLE 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

References