Part:BBa_K3271024:Design
Gal4::His3-EX-RFP-SP pSB1K3 Type IIS Assembly
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found at 3304
Illegal suffix found at 3360 - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 3304
Illegal NheI site found at 2982
Illegal SpeI site found at 3361
Illegal PstI site found at 3375
Illegal NotI site found at 3310
Illegal NotI site found at 3368 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 3304
Illegal BglII site found at 2873
Illegal BglII site found at 2933
Illegal XhoI site found at 1026
Illegal XhoI site found at 2052
Illegal XhoI site found at 3351 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found at 3304
Illegal suffix found at 3361 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found at 3304
Plasmid lacks a suffix.
Illegal XbaI site found at 3319
Illegal SpeI site found at 3361
Illegal PstI site found at 3375 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI site found at 3326
Illegal BsaI.rc site found at 3354
Design Notes
Please note the upstream and downstream Ade4 locus homologies as well as the His3 auxotrophic selection marker are found outside the prefix and suffix, not within the multiple cloning site. This contribution consists of an updated pSB1K3 BioBrick backbone that makes the BioBrick plasmids compatible with cloning in both E. coli and S. cerevisiae. This plasmid backbone is compatible with traditional and Type IIS cloning.
Source
The Gal4 upstream and downstream homologies are comprised of 100 bp each from the promoter region and the terminator region of the Gal4 locus of S. cerevisiae, respectively. The His3 auxotrophic selection marker is identical in sequence to the endogenous His3 locus of S. cerevisiae. The His3 auxotrophic selection marker is under the control of the endogenous His3 promoter and the ADH1 terminator. The RFP cassette is identical in sequence to BBa_J04450.