Plasmid

Part:BBa_J72008:Design

Designed by: John Anderson   Group: Anderson Lab   (2008-07-16)

phi80 integration helper plasmid pInt80-649


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 878
    Illegal XbaI site found at 2279
    Illegal SpeI site found at 939
    Illegal SpeI site found at 4519
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 878
    Illegal NheI site found at 884
    Illegal SpeI site found at 939
    Illegal SpeI site found at 4519
    Illegal NotI site found at 4838
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 878
    Illegal BglII site found at 2722
    Illegal BglII site found at 3416
    Illegal BamHI site found at 1263
    Illegal XhoI site found at 1576
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 878
    Illegal XbaI site found at 2279
    Illegal SpeI site found at 939
    Illegal SpeI site found at 4519
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 878
    Illegal XbaI site found at 2279
    Illegal SpeI site found at 939
    Illegal SpeI site found at 4519
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 5484


Design Notes

N/A


Source

Constructed from CRIM integration plasmid pAH123.  See ACCESSION AY048726 and PMID: 11591683 for details.
IPCR ca648F/R on pAH123		(~5200 bp, BglII/XbaI)
PCR ca649F/R on BW23474 gen	(1154 bp, BamHI/XbaI)
Ligate, product is pInt80-649	(6335 bp)
----
ca648F	Forward BglII/BsaI PCR of pAH helpers
ggagaGGTCTCagatctATATGTAACGGTGAACAGTTG
ca648R	Reverse XbaI/BsaI PCR of pAH helpers
ctgaaGGTCTCtctagaCAAAGGGAAAACTGTCCATACC
ca649F	Forward XbaI PCR of pir-116
CAGTGtctagaCATGAGTGGATAGTACGTTGC
ca649R	Reverse BamHI PCR of pir-116
CTCAAAggatccGCGTTTAACCTCTGGCTTAC

References