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Composite

Part:BBa_K1045015:Design

Designed by: iGEM Team Göttingen 2013   Group: iGEM13_Goettingen   (2013-09-20)

RBS BBa_B0034 with inversed Pre- and Suffix- Promoter reverse - Promoter - DarR operator - BBa_E0240


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 21
    Illegal NheI site found at 44
    Illegal NheI site found at 124
    Illegal NheI site found at 147
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 75
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 845


Design Notes

The inverse RBS was generated by hybridizing oligos to a double-stranded DNA fragment that corresponded to the inverse RBS sequence cut with EcoRI and SpeI at the prefix and suffix sequence. Next, this hybridization product was cloned in a prefixing composition into BBa_K1045014.

Source

The hybridization oligos mentioned under "Design Notes" were purchased from Sigma-Aldrich. The sequence information of the RBS originated from the Parts Registry page of BBa_B0034. In addition, to construct the composite part BBa_K1045015, BBa_K1045014 was used. BBa_K1045014 was constructed using hybridization oligos (sequence information: Zhang et al., 2013 and parts registry) from Sigma-Aldrich and parts taken from the distribution kit 2013.

References

Lei Zhang, Weihui Li, and Zheng-Guo He (2013) “DarR, a TetR-like Transcriptional Factor, Is a Cyclic Di-AMP-responsive Repressor in Mycobacterium smegmatis”, THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 288, NO. 5, pp. 3085–3096