Part:BBa_K2739009:Design
Hybrid promoter-PhaCAB-Bktb
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 979
Illegal BglII site found at 1804 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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.
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)