Difference between revisions of "Part:BBa K1773016"
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<partinfo>BBa_K1773016 short</partinfo> | <partinfo>BBa_K1773016 short</partinfo> | ||
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− | + | ===Construction=== | |
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− | + | This part was constructed using overlapping oligonucleotides using PCR. | |
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+ | fw oligo: gctgaattcgcggccgcttctagagacctgtaggatcgtacaggtttacgcaagaaaatggttt | ||
+ | |||
+ | rev oligo: ggcACTAGTtttctcctctttaattatgcatgtgtatcaccgccagaggtattcgactataacaaaccattttcttgcgtaa | ||
+ | |||
+ | 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_K1773017 and BBa_K1773018 | ||
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+ | Later these parts were sequence for confirmation. | ||
+ | |||
+ | [[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]] |
Revision as of 15:34, 11 September 2015
pLux/cI right promoter with strong RBS
pLux/cI promoter with strong RBS.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Construction
This part was constructed using overlapping oligonucleotides using PCR.
fw oligo: gctgaattcgcggccgcttctagagacctgtaggatcgtacaggtttacgcaagaaaatggttt
rev oligo: ggcACTAGTtttctcctctttaattatgcatgtgtatcaccgccagaggtattcgactataacaaaccattttcttgcgtaa
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_K1773017 and BBa_K1773018
Later these parts were sequence for confirmation.