Part:BBa_K1807007:Design
Synthetic Phosphate-specific Transporter Operon
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 2133
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 529
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 655
Illegal NgoMIV site found at 716
Illegal NgoMIV site found at 1732
Illegal NgoMIV site found at 2305
Illegal AgeI site found at 164
Illegal AgeI site found at 2762 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1417
Illegal BsaI site found at 2383
Illegal SapI site found at 1223
Illegal SapI.rc site found at 3169
Design Notes
iGEM York 2015 generated this part by designing two overlapping G Block fragments (1900bp and 1943bp) that were used in a Gibson Assembly Reaction with BBa_K1807000.
This Phosphate-specific transporter operon has been designed and re-factored (ribosome-binding sites have been introduced between each gene). Each protein domain has been based on amino acid sequence from a metagenomic sequence. The operon coding sequences have been codon-optimized for Escherichia coli K-12 translation.
Notes on Growth
A noticeable slowing of the growth rate was observed in DH5 alpha cells even when transcription was not induced (grown on LB-Agar with appropriate antibiotic). This may be due to leaky expression (WT lacI is present in DH5 alpha rather than lacIq) increasing the translational energetic demand on the cells. It is also possible (but not confirmed) that the protein has a toxic effect on cellular metabolism.
Source
Candidatus Accumulibacter phosphatis SK-1 strain