Generator

Part:BBa_K1758350:Design

Designed by: Team Bielefeld-CeBiTec 2015   Group: iGEM15_Bielefeld-CeBiTec   (2015-09-08)

Nickel repressor under control of constitutive promoter and strong RBS


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We used the codon optimized, synthesized Sequence of rcnR from E.coli K12 under the control of a constitutive Primer( BBa_K608002). We designed the used primes, in specially split primers, for Gibson Assembly to ligate the constitutive Promoter and rcnR with pSB1C3. For this aim we designed four primers for the generation of homologous overlaps between the synthesized constitutive promoter+rcnR and the pSB1C3:

cm_fwd

CGGCATCAGCACCTTGTC

pSB1C3_rcnR_rev

GAAAGTCCTTGATTCGTACATCAAATGACTCTAGAAGCGGCCGCGAAT

cm_rev

TATACGCAAGGCGACAAG

pSB1C3_kPrm_fwd

CTAGGACTGAGCTAGCTGTCAATACTAGTAGCGGCCGCTGCA


Source

Synthesized, codon optimized gene of E. coli Synthesized by IDT


References

Synthesized by IDT