Coding

Part:BBa_K2725013

Designed by: Eva Steele   Group: iGEM18_Edinburgh_UG   (2018-10-11)


P1003 Casette with 10 Amber substitutions

P1003 Kanamycin resistance cassette with 10 serine codons in the coding sequence substituted for amber stop codons (TAG): S17Amb, S36Amb, S60Amb, S116Amb, S133Amb, S154Amb, S189Amb, S200Amb, S240Amb, S242Amb. When transformed into E .coli there is no P1003 expression and no kanamycin resistance showing that 10 amber substitutions effectively prevents expression. This part is an improvement upon BBa_K914018 (http://parts.igem.org/Part:BBa_K914018)

T--Edinburgh_UG--400.png

Figure 1. Growth curves of double transformed Top10 E. coli strains in 400 μg/mL Kanamycin.

T--Edinburgh_UG--prob_per_stop.png

Figure 2. Graph showing the probability of a read-through (and therefore expression) of a gene with number of amber STOP codons within the coding sequence of the gene. When there are 10 amber STOP codons in the coding sequence 1 in every 10^18 translation attempts may result in expression of the gene.

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]


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Parameters
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