Difference between revisions of "Part:BBa K249004"
 
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This part was designed as a plasmid vector backbone and was expected to work by adding an N-terminal Arginine Tag to the protein to which it would be fused.
 
This part was designed as a plasmid vector backbone and was expected to work by adding an N-terminal Arginine Tag to the protein to which it would be fused.
  
−
This part was shown to work via 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.
 +
<br><br>
  
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pSB-N-terminus
+
<html>
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Score = 60.0 bits (30), Expect = 1e-15
+
<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 10 (The vector sent in)
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<tr>
−
ATGCGCCGCCGCCGCCGCCGCCGCCGCCGC 39
+
<th>
 +
<img src="https://static.igem.org/mediawiki/parts/3/3e/UofLNvsC-YFP1.jpg" width="450"/>
 +
</th>
  
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Sbjct 1 (the sequence expected)
+
<th>
−
ATGCGCCGCCGCCGCCGCCGCCGCCGCCGC 30
+
<img src="https://static.igem.org/mediawiki/parts/6/62/UofLNvsC-YFP2.jpg" width="450"/>
 +
</th>
 +
</tr>
  
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We fused a YFP gene with an N-terminal (Biofusion standard prefix)
+
</table>
 +
</body>
 +
</center>
 +
</html>
  
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mRBS-N-terminal EYFP
 
−
Score = 1379 bits (717), Expect = 0.0
 
−
Identities = 717/717 (100%), Gaps = 0/717 (0%)
 
−
Strand=Plus/Plus
 
−
 
−
Query 147 (The vector sent in)
 
−
ATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGAC 206
 
−
 
−
Sbjct 1 (the sequence expected)
 
−
ATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGAC 60
 
−
 
−
 
−
Query 207 GGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTAC 266
 
−
Sbjct 61  GGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTAC 120
 
−
 
−
 
−
Query 267 GGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACC 326
 
−
Sbjct 121 GGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACC 180
 
−
 
−
 
−
Query 327 CTCGTGACCACCTTCGGCTACGGCCTGCAATGCTTCGCCCGCTACCCCGACCACATGAAG 386
 
−
Sbjct 181 CTCGTGACCACCTTCGGCTACGGCCTGCAATGCTTCGCCCGCTACCCCGACCACATGAAG 240
 
−
 
−
 
−
Query 387 CTGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTC 446
 
−
Sbjct 241 CTGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTC 300
 
−
 
−
 
−
Query 447 TTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTG 506
 
−
Sbjct 301 TTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTG 360
 
−
 
−
 
−
Query 507 GTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCAC 566
 
−
Sbjct 361 GTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCAC 420
 
−
 
−
 
−
Query 567 AAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAAC 626
 
−
Sbjct 421 AAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAAC 480
 
−
 
−
 
−
Query 627 GGCATCAAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCC 686
 
−
Sbjct 481 GGCATCAAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCC 540
 
−
 
−
 
−
Query 687 GACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCAC 746
 
−
Sbjct 541 GACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCAC 600
 
−
 
−
 
−
Query 747 TACCTGAGCTACCAGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTC 806
 
−
Sbjct 601 TACCTGAGCTACCAGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTC 660
 
−
 
−
 
−
Query 807 CTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAG 863
 
−
Sbjct 661 CTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAG 717
 
−
 
−
The Part YFP part which was designed carried the N-terminal fusion tag.
 
  
  

Latest revision as of 20:35, 27 October 2010

N-terminal Arginine fusion vector

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

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 vector backbone and was expected to work by adding an N-terminal Arginine Tag to the protein to which it would be fused.

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.