Plasmid_Backbone

Part:BBa_K394001:Design

Designed by: Brian Landry   Group: iGEM10_WashU   (2010-10-20)

Plasmid for Chromosomal Integration in Yeast at His3


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3436
    Illegal NheI site found at 1267
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 3442
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3436
    Illegal BamHI site found at 1413
    Illegal XhoI site found at 1032
    Illegal XhoI site found at 2315
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 3436
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 3436
    Plasmid lacks a suffix.
    Illegal XbaI site found at 3451
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NgoMIV site found at 1439
    Illegal NgoMIV site found at 1807
    Illegal NgoMIV site found at 1967
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 2461


Design Notes

None


Source

This part was derived from plasmid pSB1AT3. Two successive rounds of PCR inserted a BbsI cut site and 40 bp homologous region in before the prefix and after the suffix.

The following Primers were used: 

Forward 1	GCACAGAACAATAACCTGCTGGAAACGAAGATAAATCgaagacGATTACTTCGCGTTATGCAGGC 
Reverse 1	GCATCTTCTCAAATATGCTTCCCAGCCTGCTTATCcttctgAAATTCTGCCTCGTGATACGCC
Forward 2	tactagtagcggccgctgcagTCTTAACCCAACTGCACAGAACAATAACCTGCTGGAAACG
Reverse 2	ctctagaagcggccgcgaattcTTAGTATTGCTGGCCGCATCTTCTCAAATATGCTTCCCAGCC

References