Device
Part:BBa_K1465306:Design
Designed by: iGEM-Team Bielefeld 2014 Group: iGEM14_Bielefeld-CeBiTec (2014-10-08)
Isobutanol pathway under Ptac control
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 6916
Illegal XhoI site found at 6196 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 3579
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 3165
Illegal BsaI site found at 5982
Illegal BsaI site found at 6239
Illegal BsaI.rc site found at 1567
Illegal BsaI.rc site found at 2161
Illegal BsaI.rc site found at 3984
Design Notes
Coding sequences
We started with the pSB1C3_alsS_ilvC_ilvD_kivD construct which can be found in the parts registry as BBa_K1465302. Therefore we used the CDS of the NCTU team Formosa 2011/2012 BioBricks alsS (BBa_K539627), ilvC (BBa_K539621),
ilvD (BBa_K539626) and kivD (BBa_K539742).
For the characterization we wanted to add the Ptac promoter (BBa_K731500) to our construct.
We tried BioBrick Suffix Assembly where the Ptac was cut out of the gel and BioBrick Prefix Assembly where we cut the alsS_ilvC_ilvD_kivD part out of the gel. In the end, the suffix version worked out and we created the BioBrick device pSB1C3_Ptac_alsS_ilvC_ilvD_kivD
Source
For this device we used the following BioBricks: