Part:BBa_K3271022:Design
Ade4::Ura3-EX-RFP-SP pSB1K3 Type IIS
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found at 3530
Illegal suffix found at 3586 - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 3530
Illegal SpeI site found at 3587
Illegal PstI site found at 3601
Illegal NotI site found at 3536
Illegal NotI site found at 3594 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 3530
Illegal XhoI site found at 1026
Illegal XhoI site found at 2052
Illegal XhoI site found at 3577 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found at 3530
Illegal suffix found at 3587 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found at 3530
Plasmid lacks a suffix.
Illegal XbaI site found at 3545
Illegal SpeI site found at 3587
Illegal PstI site found at 3601 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI site found at 3212
Illegal BsaI site found at 3552
Illegal BsaI.rc site found at 3580
Illegal SapI.rc site found at 3059
Design Notes
Please note the upstream and downstream Ade4 locus homologies as well as the Ura3 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 Ade4 upstream and downstream homologies are comprised of 100 bp each from the promoter region and the terminator region of the Ade4 locus of S. cerevisiae, respectively. The Ura3 selection marker is identical in sequence to that of the Ura3 locus in S. cerevisiae. The Ura3 auxotrophic selection marker is under the control of the endogenous Ura3 promoter and the ADH1 terminator. The RFP cassette is identical in sequence to BBa_J04450.