Regulatory

Part:BBa_K1773016

Designed by: Sarunas Tumas   Group: iGEM15_Vilnius-Lithuania   (2015-08-25)

pLux/cI right promoter with strong RBS

pLux/cI promoter with strong 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: 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.

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.
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