Difference between revisions of "Part:BBa J72008:Design"

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Constructed from CRIM integration plasmid pAH123.  See ACCESSION AY048726 and PMID: 11591683 for details.
 
Constructed from CRIM integration plasmid pAH123.  See ACCESSION AY048726 and PMID: 11591683 for details.
−
PCR ca648F/R on pAH123 (~5200 bp, BglII/XbaI)
+
IPCR ca648F/R on pAH123 (~5200 bp, BglII/XbaI)
 
PCR ca649F/R on BW23474 gen (1154 bp, BamHI/XbaI)
 
PCR ca649F/R on BW23474 gen (1154 bp, BamHI/XbaI)
 
Ligate, product is pInt80-649 (6335 bp)
 
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
 
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===References===
 
===References===

Latest revision as of 21:15, 16 July 2008

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