Difference between revisions of "Part:BBa K2996002"
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<partinfo>BBa_K2996002 short</partinfo>
 
<partinfo>BBa_K2996002 short</partinfo>
  
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The arrays consist of 69 bp of the respective leader, including the sequence elements essential for adaptation, and one spacer flanked by two repeats. Wildtype E.coli has two types of CRISPRER array, with their repeat sequence differ in one base pair. Thus, we name them 1T and 2A.
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The cassette consists of 69 bp of the respective leader, including the sequence elements essential for adaptation, and one spacer flanked by two repeats. Wildtype E.coli has two types of CRISPRER array, with their repeat sequence differ in one base pair. Thus, we name them 1T and 2A.
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Following RSRL, there is a fluorescent protein gene, EGFP. Originally, there is a stop codon within the RSRL sequence and the fluorescent protein gene was not in the correct reading frame, so there would be no fluorescent protein expressed. After illuminating, Cas1/2 would insert 61bp protospacer and another repeat sequence, which would result in frame shifting. In that case, the stop codon would disappear, and EGFP enter the correct reading frame.
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When you want to read the information, inducer should be added and expression of fluorescent protein would be activated.  
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===Usage and Biology===
 
===Usage and Biology===
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In our design, this integration cassette RSRL in pRead consists of a leader and two repeats interspaced by a spacer fused downstream, to the complete coding sequence of an out-of-frame EGFP gene (EGFP +2).
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Originally, there is a stop codon within the RSRL sequence and the fluorescent protein gene was not in the correct reading frame, so there would be no fluorescent protein expressed. After illuminating, Cas1/2 would insert 61bp protospacer and another repeat sequence, which would result in frame shifting. In that case, the stop codon would disappear, and EGFP enter the correct reading frame.
 +
 +
When you want to read the information, inducer should be added and expression of fluorescent protein would be activated.
 +
 +
 +
  
 
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Revision as of 12:21, 4 October 2019


minimal CRISPR array-A

The cassette consists of 69 bp of the respective leader, including the sequence elements essential for adaptation, and one spacer flanked by two repeats. Wildtype E.coli has two types of CRISPRER array, with their repeat sequence differ in one base pair. Thus, we name them 1T and 2A.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]