Difference between revisions of "Part:BBa K4808011"
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     <p> Step1: Transformation of the pEcCas plasmid(carry cas9 protein) into E. coli AIS-0 (CICC20905). Step2:  construction of the pTarget plasmid and donor DNA. Step3:  transformation of pTarget plasmid and donor DNA into the AIS-0 with pEsCas inserted. Step4: Incubation of the transformed strains. </p >
 
     <p> Step1: Transformation of the pEcCas plasmid(carry cas9 protein) into E. coli AIS-0 (CICC20905). Step2:  construction of the pTarget plasmid and donor DNA. Step3:  transformation of pTarget plasmid and donor DNA into the AIS-0 with pEsCas inserted. Step4: Incubation of the transformed strains. </p >
  

Revision as of 14:10, 12 October 2023

pEcCas

pEcCas plasmid carry the cas 9 protein which can cut the target gene.

Characterization

pEcCas plasmid play an important role in our CRISPR-CAS 9 knocking out experiment.

https://static.igem.wiki/teams/4808/wiki/.png

Step1: Transformation of the pEcCas plasmid(carry cas9 protein) into E. coli AIS-0 (CICC20905). Step2: construction of the pTarget plasmid and donor DNA. Step3: transformation of pTarget plasmid and donor DNA into the AIS-0 with pEsCas inserted. Step4: Incubation of the transformed strains.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 4334
    Illegal EcoRI site found at 8435
    Illegal EcoRI site found at 9944
    Illegal EcoRI site found at 11128
    Illegal EcoRI site found at 12594
    Illegal XbaI site found at 7132
    Illegal SpeI site found at 1124
    Illegal PstI site found at 1805
    Illegal PstI site found at 2760
    Illegal PstI site found at 9439
    Illegal PstI site found at 9686
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4334
    Illegal EcoRI site found at 8435
    Illegal EcoRI site found at 9944
    Illegal EcoRI site found at 11128
    Illegal EcoRI site found at 12594
    Illegal NheI site found at 4093
    Illegal NheI site found at 14303
    Illegal SpeI site found at 1124
    Illegal PstI site found at 1805
    Illegal PstI site found at 2760
    Illegal PstI site found at 9439
    Illegal PstI site found at 9686
    Illegal NotI site found at 1443
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4334
    Illegal EcoRI site found at 8435
    Illegal EcoRI site found at 9944
    Illegal EcoRI site found at 11128
    Illegal EcoRI site found at 12594
    Illegal BglII site found at 13274
    Illegal BamHI site found at 6372
    Illegal BamHI site found at 8374
    Illegal XhoI site found at 2613
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 4334
    Illegal EcoRI site found at 8435
    Illegal EcoRI site found at 9944
    Illegal EcoRI site found at 11128
    Illegal EcoRI site found at 12594
    Illegal XbaI site found at 7132
    Illegal SpeI site found at 1124
    Illegal PstI site found at 1805
    Illegal PstI site found at 2760
    Illegal PstI site found at 9439
    Illegal PstI site found at 9686
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 4334
    Illegal EcoRI site found at 8435
    Illegal EcoRI site found at 9944
    Illegal EcoRI site found at 11128
    Illegal EcoRI site found at 12594
    Illegal XbaI site found at 7132
    Illegal SpeI site found at 1124
    Illegal PstI site found at 1805
    Illegal PstI site found at 2760
    Illegal PstI site found at 9439
    Illegal PstI site found at 9686
    Illegal AgeI site found at 8209
    Illegal AgeI site found at 9866
    Illegal AgeI site found at 13006
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 8191