Difference between revisions of "Part:BBa K4808009"
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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 >
  
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K4808009 SequenceAndFeatures</partinfo>
  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K4808009 parameters</partinfo>
 
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Revision as of 10:42, 11 October 2023

pTarget

pTarget plasmid that carrying specific gRNA sequence which can identity the target gene for the further gene knockout.

Characterization

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

1280x1280.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 330
    Illegal XbaI site found at 336
    Illegal SpeI site found at 221
    Illegal PstI site found at 348
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 330
    Illegal NheI site found at 198
    Illegal SpeI site found at 221
    Illegal PstI site found at 348
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 330
    Illegal BglII site found at 360
    Illegal BamHI site found at 186
    Illegal XhoI site found at 384
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 330
    Illegal XbaI site found at 336
    Illegal SpeI site found at 221
    Illegal PstI site found at 348
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 330
    Illegal XbaI site found at 336
    Illegal SpeI site found at 221
    Illegal PstI site found at 348
    Illegal NgoMIV site found at 1155
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 122