Plasmid

Part:BBa_K1160001

Designed by: Yang Rongwu   Group: iGEM13_NJU_NJUT_China   (2013-09-14)


chimeric RNA，combining crRNA and tracrRNA

CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system provide protection against mobile genetic element by guided specific DNA cleavage. Cas9 protein belongs type II CRISPR system and showed strong DNA cleavage activity. In order to accomplish site specific cleavage, cas9 protein must be guided by crRNA and tracrRNA. This plasmid carries the fragment corresponding to a chimeric RNA that combines crRNA and tracrRNA. The crRNA is designed to target part BBa_K1160002, which is used to examine the DNA cleavage conducted by Cas9 protein or detect the existence of CRISPR system in other organism. The crRNA segment is replaceable，and it's flanked by BsaA I site. (both upstream and down stream)

You can treat this part with BasA I and ligate it with your seed sequence (your seed sequence must also be flanked by BasA I sticky end).

NOTE：crRNA contains the sequence that is complementary to your target sequence.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 249
    Illegal XbaI site found at 1427
    Illegal SpeI site found at 712
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 249
    Illegal SpeI site found at 712
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 249
    Illegal BglII site found at 1412
    Illegal BamHI site found at 969
    Illegal BamHI site found at 1689
    Illegal XhoI site found at 260
    Illegal XhoI site found at 450
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 249
    Illegal XbaI site found at 1427
    Illegal SpeI site found at 712
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 249
    Illegal XbaI site found at 1427
    Illegal SpeI site found at 712
    Illegal NgoMIV site found at 1442
    Illegal NgoMIV site found at 1813
    Illegal NgoMIV site found at 3882
    Illegal AgeI site found at 137
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 156
    Illegal BsaI site found at 4895
    Illegal BsaI.rc site found at 143
    Illegal BsaI.rc site found at 3988
    Illegal SapI site found at 1822


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Categories
Parameters
None