Composite

Part:BBa_K1045014:Design

Designed by: iGEM Team Göttingen 2013   Group: iGEM13_Goettingen   (2013-09-20)
Revision as of 19:43, 4 October 2013 by Kati (Talk | contribs) (→‎Source)

Promoter reverse - Promoter - DarR operator - GFP generator BBa_E0240


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1
    Illegal NheI site found at 24
    Illegal NheI site found at 104
    Illegal NheI site found at 127
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 55
    Illegal AgeI site found at 963
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 825


Design Notes

This part was constructed using hybridization oligos for BBa_K1045011. The hybridization product corresponded to a DNA fragment harboring the sequence of BBa_K1045011 cut with EcoRI and SpeI at the prefix and suffix sites. This fragment was ligated into BBa_K1045013 in a prefixing composition.

This biobrick harbors a mutation in the suffix sequence. The mutated suffix sequence is: TACTAGTAACGGCCGCTGCAG. This eliminates the NotI restriction site. The plasmid might still be cut with SpeI and PstI.

Source

The hybridization oligos mentioned under "Design Notes" were purchased from Sigma-Aldrich. The sequence of the promoter originated from the Parts Registry page of BBa_J23117. The 54 bp upstream of the promoter represent a random sequence which would translate into "cyclicdiampacterim". To construct this composite part, BBa_K0145013 was used. It consisted of another hybridization oligo BBa_K0145012 and the part BBa_E0240, which derived from the distribution kit 2013.

References