Difference between revisions of "Part:BBa K2729009"
 
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<partinfo>BBa_K2729009 short</partinfo>
 
<partinfo>BBa_K2729009 short</partinfo>
  
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AmCyan is one of the existing violet fluorochromes for use in flow cytometers equipped with a violet (405 nm) laser. FMDV 2A is used widely for this purpose since multiple proteins can be coexpressed by linking open reading frames (ORFs) to form a single cistron.
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AmCyan is one of the existing violet fluorochromes for use in flow cytometers equipped with a violet (405 nm) laser. FMDV 2A is "self-cleaving" peptides, and upstream coding region can be a single cistron with the assistance.
  
 
===Characterization===
 
===Characterization===
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The sequence is optimized for <i>Homo Sapiens</i> using IDT Codon Optimization Tool. Our team checked the sequence length of synthesized oligo DNA and characterized it.
 
The sequence is optimized for <i>Homo Sapiens</i> using IDT Codon Optimization Tool. Our team checked the sequence length of synthesized oligo DNA and characterized it.
  
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[[File:T--Tokyo_Tech--test-electrop.gif|thumb|left|300px| '''Figure 1:''' '''Result of the qualitative experiment''' - description comes here. ]]
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[[File:Tokyotech electrophoresis.png|thumb|left|300px| '''Figure 1:''' '''Result of the Electrophoresis(Marker: lambda HindIII)''' ]]
  
 
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As shown in Figure 1, the band comes between 900 bp to 1,000 bp.  
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As shown in Figure 1, the band comes between 900 bp to 1,000 bp. The actual length without prefix and suffix is 920 bp.
 
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<br style="clear: both" />
  
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Our team successfully validated that the sequence was synthesized, and obtained enough amount of it for transfection.
 
Our team successfully validated that the sequence was synthesized, and obtained enough amount of it for transfection.
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The function of FMDV2a will be tested by testing the infection of pseudo-virus into Vero cells. If the 2a peptide works well, the fluorescence intensity can be observed robustly.
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===Sequence and Features===
 
===Sequence and Features===
 
<partinfo>BBa_K2729009 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2729009 SequenceAndFeatures</partinfo>
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===Reference===
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- [https://www.takarabio.com/products/gene-function/fluorescent-proteins/fluorescent-protein-plasmids/cyan-and-green-fluorescent-proteins/amcyan1-fluorescent-protein Takara Bio - AmCyan1 fluorescent protein]

Latest revision as of 03:24, 18 October 2018


AmCyan_FMDV2a

AmCyan is one of the existing violet fluorochromes for use in flow cytometers equipped with a violet (405 nm) laser. FMDV 2A is "self-cleaving" peptides, and upstream coding region can be a single cistron with the assistance.

Characterization

The sequence is optimized for Homo Sapiens using IDT Codon Optimization Tool. Our team checked the sequence length of synthesized oligo DNA and characterized it.

Figure 1: Result of the Electrophoresis(Marker: lambda HindIII)


As shown in Figure 1, the band comes between 900 bp to 1,000 bp. The actual length without prefix and suffix is 920 bp.

Analysis

Our team successfully validated that the sequence was synthesized, and obtained enough amount of it for transfection.

The function of FMDV2a will be tested by testing the infection of pseudo-virus into Vero cells. If the 2a peptide works well, the fluorescence intensity can be observed robustly.


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 43
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 820
    Illegal AgeI site found at 669
  • 1000
    COMPATIBLE WITH RFC[1000]

Reference

- Takara Bio - AmCyan1 fluorescent protein