Difference between revisions of "Part:BBa K1773017"

 
 
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__NOTOC__
 
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<partinfo>BBa_K1773017 short</partinfo>
 
<partinfo>BBa_K1773017 short</partinfo>
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<partinfo>BBa_K1773017 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1773017 SequenceAndFeatures</partinfo>
  
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===Construction===
  
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<!-- Uncomment this to enable Functional Parameter display
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This part was constructed using overlapping oligonucleotides using PCR.
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===Functional Parameters===
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<partinfo>BBa_K1773017 parameters</partinfo>
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fw oligo: gctgaattcgcggccgcttctagagacctgtaggatcgtacaggtttacgcaagaaaatggttt
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rev oligo: gcaACTAGTctttcctgtgtgatatgcatgtgtatcaccgccagaggtattcgactataacaaaccattttcttgcgtaa
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Next the PCR products were digested with EcoRI and SpeI, and ligated to predigested pSB1C3 vector. Colony PCR of four colonies was executed using VF2 and VR primers (Figure 1), and a successfull cloning was confirmed. Note: the gel also shows colony PCR of BBa_K1773016 and BBa_K1773018
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Later these parts were sequence for confirmation.
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[[File:Promot su RBS1.png|300px|thumb|'''Figure1. Colony PCR.''' M - O'Gene DNA ladder mix DNA ladder; Four colonies of each pLux/cI promoter with Strong RBS (SRBS), Medium RBS (MRBS) and Weak RBS (WRBS) sites were analysed. |center]]

Latest revision as of 15:40, 11 September 2015

pLux/cI right promoter with medium RBS

pLux/cI right promoter with medium RBS.

Sequence and Features


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]

Construction

This part was constructed using overlapping oligonucleotides using PCR.

fw oligo: gctgaattcgcggccgcttctagagacctgtaggatcgtacaggtttacgcaagaaaatggttt

rev oligo: gcaACTAGTctttcctgtgtgatatgcatgtgtatcaccgccagaggtattcgactataacaaaccattttcttgcgtaa


Next the PCR products were digested with EcoRI and SpeI, and ligated to predigested pSB1C3 vector. Colony PCR of four colonies was executed using VF2 and VR primers (Figure 1), and a successfull cloning was confirmed. Note: the gel also shows colony PCR of BBa_K1773016 and BBa_K1773018

Later these parts were sequence for confirmation.

Figure1. Colony PCR. M - O'Gene DNA ladder mix DNA ladder; Four colonies of each pLux/cI promoter with Strong RBS (SRBS), Medium RBS (MRBS) and Weak RBS (WRBS) sites were analysed.