Difference between revisions of "Part:BBa K249005"
 
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This part was designed as a plasmid backbone to allow for easy attachment of a C-terminal Arginine TAg to a gene of choice for localization into the lumazine Synthase microcompartment.
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This part was designed as a plasmid backbone to allow for easy attachment of a C-terminal Arginine Tag to a gene of choice for localization into the Lumazine Synthase microcompartment.
  
 
This part was shown to work vis the sequencing information that we received.
 
This part was shown to work vis the sequencing information that we received.
  
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pSB - C-terminus
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N-terminal tagged YFP did not have substantially more fluorescence than control cells. Cells expressing C-terminal tagged YFP had ten times more fluorescence than control cells and cells expressing N-terminal tagged YFP.
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>lcl|20071
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<br><br>
  
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Length=30
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<html>
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Score = 60.0 bits (30), Expect = 1e-15
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<center>
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Identities = 30/30 (100%), Gaps = 0/30 (0%)
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<body>
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Strand=Plus/Plus
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<table border="0" cellpadding="8"  width="28%">
  
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Query 12 (the plasmid sent in)
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<tr>
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CGCCGCCGCCGCCGCCGCCGCCGCCGCTAA 41
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<th>
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<img src="https://static.igem.org/mediawiki/parts/3/3e/UofLNvsC-YFP1.jpg" width="450"/>
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</th>
  
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Sbjct 1 (the sequence expected)
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<th>
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CGCCGCCGCCGCCGCCGCCGCCGCCGCTAA 30
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<img src="https://static.igem.org/mediawiki/parts/6/62/UofLNvsC-YFP2.jpg" width="450"/>
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</th>
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</tr>
  
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We fused a YFP gene with a C-terminal BioFusion standar suffix to this gene
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</table>
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</body>
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C-terminal EYFP-dT
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</center>
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Score = 19.9 bits (10), Expect = 0.80
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</html>
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Identities = 10/10 (100%), Gaps = 0/10 (0%)
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Strand=Plus/Plus
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Query 659 (the plasmid sent in)
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GAAGTTCATC 668
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Sbjct 135 (the sequence expected)
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GAAGTTCATC 144
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Query 714 ACAGCCACA 722
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Sbjct 440 ACAGCCACA 448
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The part was sequenced and showed some gaps due to low resolution of the sequencing data. However, it does show that the YFP gene is present, as is the C-terminal Arginine tag. Theses are simply examples of positive hits that came back, although there are many more that are present. In all there were 10 gaps in the sequence, all of which corresponded to N nucleotides in the sequencing information we received.
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K249005 parameters</partinfo>
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<!-- -->
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Latest revision as of 20:32, 27 October 2010

C-terminal Arginine Fusion Vector

pSB1A3 with an C-terminal Arginine tag. This particular plasmid utilizes the BioFusion (aka Silver Lab) Standard for BioBrick assembly. Adds 10 Arginine residues to the C-terminus of a Protein (Includes Stop Codon)

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.


This part was designed as a plasmid backbone to allow for easy attachment of a C-terminal Arginine Tag to a gene of choice for localization into the Lumazine Synthase microcompartment.

This part was shown to work vis the sequencing information that we received.

N-terminal tagged YFP did not have substantially more fluorescence than control cells. Cells expressing C-terminal tagged YFP had ten times more fluorescence than control cells and cells expressing N-terminal tagged YFP.