Difference between revisions of "Part:BBa K2739009:Design"
 
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===References===
 
===References===
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1. Poehlein,A., Kusian,B., Friedrich,B., Daniel,R. and Bowien,B. (2011) Complete genome sequence of the type strain Cupriavidus necator N-1. J. Bacteriol. 193 (18)

Latest revision as of 20:33, 16 October 2018


Hybrid promoter-PhaCAB-Bktb


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
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 979
    Illegal BglII site found at 1804
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 285
    Illegal NgoMIV site found at 356
    Illegal NgoMIV site found at 956
    Illegal NgoMIV site found at 1268
    Illegal NgoMIV site found at 1547
    Illegal NgoMIV site found at 2199
    Illegal NgoMIV site found at 2221
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4065
    Illegal BsaI site found at 5108


Design Notes

This is a composite part, which developed based on the BBa_K2739008 (Btkb) and BBa_K1149051 (Hybrid-promoter-phaCAB).

For the Bktb part, its length is 1185bp and excess the 1kb length limit of the IDT sponsor limitation to be generated. Therefore two overlapped fragments of Bktb were then designed and generated by IDT. A set of PCR primer were then design to generate the whole Bktb through the PCR amplification. T--Edinburgh_OG--Notebook_-_bktb_owen1.png

Figure 1. B1 and B2 fragments were codon-optimised for E. coli and synthesised by IDT. The 696 bp overlap was in green and restriction enzyme sites were coloured by red. Two pairs of primers were ordered from IDT to amplify bktB, among which B1 forward primer and B2 reverse primer were used to amplify bktB fragments.


Source

BBa_K2739008 (Btkb) used in the part was created through IDT. The original sequence would be accessed in NCBI: NC_015726.1.

Bktb sequence is codon optimised by IDT. BBa_K1149051 (Hybrid-promoter-phaCAB) is a available biobrick.

References

1. Poehlein,A., Kusian,B., Friedrich,B., Daniel,R. and Bowien,B. (2011) Complete genome sequence of the type strain Cupriavidus necator N-1. J. Bacteriol. 193 (18)