Difference between revisions of "Part:BBa K1773003:Design"
Line 9: Line 9:
 
These mutagenic primers were used:
 
These mutagenic primers were used:
  
−
EcoRI mutagenesis :
+
EcoRI mutagenesis: <br />
−
Fw: cgttttcaatggcagaatccggcatttgatttggca
+
F: cgttttcaatggcagaatccggcatttgatttggca<br />
−
Rev: tgccaaatcaaatgccggattctgccattgaaaacg
+
R: tgccaaatcaaatgccggattctgccattgaaaacg<br />
  
−
XbaI Mutagenesis:
+
XbaI Mutagenesis:<br />
−
Fw: cgatcatcattattcttctctggatgctgatgttaatttggg
+
F: cgatcatcattattcttctctggatgctgatgttaatttggg<br />
−
Rev: cccaaattaacatcagcatccagagaagaataatgatgatcg
+
R: cccaaattaacatcagcatccagagaagaataatgatgatcg<br />
  
−
PstI Mutagenesis:
+
PstI Mutagenesis:<br />
−
Fw: gaagactgaactgcctgcggttaaacaacataaac
+
F: gaagactgaactgcctgcggttaaacaacataaac<br />
−
Rev: gtttatgttgtttaaccgcaggcagttcagtcttc
+
Re: gtttatgttgtttaaccgcaggcagttcagtcttc<br />

Revision as of 15:51, 9 September 2015

I-F type CRISPR-Cas Cas3 gene


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Unknown
  • 12
    INCOMPATIBLE WITH RFC[12]
    Unknown
  • 21
    INCOMPATIBLE WITH RFC[21]
    Unknown
  • 23
    INCOMPATIBLE WITH RFC[23]
    Unknown
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2307
    Illegal AgeI site found at 2680
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Gene mutagenesis was performed using Invitrogen Gene-art multi site mutagenesis PLUS kit.

These mutagenic primers were used:

EcoRI mutagenesis:
F: cgttttcaatggcagaatccggcatttgatttggca
R: tgccaaatcaaatgccggattctgccattgaaaacg

XbaI Mutagenesis:
F: cgatcatcattattcttctctggatgctgatgttaatttggg
R: cccaaattaacatcagcatccagagaagaataatgatgatcg

PstI Mutagenesis:
F: gaagactgaactgcctgcggttaaacaacataaac
Re: gtttatgttgtttaaccgcaggcagttcagtcttc