Composite

Part:BBa_K3120016

Designed by: Ruoyi Xue   Group: iGEM19_BM-AMU   (2019-08-14)


pU6+gRNA2+pCAG+hspCas9+AmpR promoter+AmpR

U6 promoter for transcription of downstream gRNA sequences. gRNA2 consists of E-cadherin's 20nt target sequence and sgRNA scaffold, which directs Cas protein localization downstream of the E-cadherin sequence. CAG promoter consists of CMV enhancer, chicken β-actin promoter and rabbit β-globin， which is used to initiate transcription and translation of downstream hspCas9 proteins. hspCas9 works with gRNA to cut a gap of the E-cadherin gene's downstream for insertion of BBa_K3120018. AmpR promoter+ and AmpR are used to screen plasmids.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1202
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2029
    Illegal NgoMIV site found at 3133
    Illegal NgoMIV site found at 3206
    Illegal NgoMIV site found at 3691
    Illegal NgoMIV site found at 4600
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 5890


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Categories
Parameters
None