Freiburg07/report fusion parts

From 2007.igem.org

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additional amino acids (A, G) are encoded before the start of the protein.</li>
additional amino acids (A, G) are encoded before the start of the protein.</li>
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<p><b><font size="4">N-part</font></b><br>
<p><b><font size="4">N-part</font></b><br>
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|&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; |&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;  
|&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; |&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;  
|&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; |&nbsp;&nbsp;&nbsp; ||&nbsp;&nbsp; |<br>
|&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; |&nbsp;&nbsp;&nbsp; ||&nbsp;&nbsp; |<br>
-
&nbsp;&nbsp;&nbsp; <u><font color="#FF00FF">GAATTC</font><font color="#009900">gcggccgc</font>t<font color="#FF0000">TCTAG</u><i>A</i></font><i>tg</i>nnnnnn<font color="#0000FF">ACCGGT</font>taa<u>t<font color="#FF0000">ACTAGT</font>a<font color="#009900">gcggccg</font><font color="#FF00FF">CTGCAG</font></u><br>
+
&nbsp;&nbsp;&nbsp; <u><font color="#FF00FF">GAATTC</font><font color="#009900">gcggccgc</font>t<font color="#FF0000">TCTAG</font></u><font color="#FF0000"><i>A</i></font><i>tg</i>nnnnnn<font color="#0000FF">ACCGGT</font>taa<u>t<font color="#FF0000">ACTAGT</font>a<font color="#009900">gcggccg</font><font color="#FF00FF">CTGCAG</font></u><br>
&nbsp; 1 ---------+---------+---------+---------+---------+--------- 59<br>
&nbsp; 1 ---------+---------+---------+---------+---------+--------- 59<br>
&nbsp;&nbsp;&nbsp; CTTAAGcgccggcgaAGATCTacnnnnnnTGGCCAattaTGATCAtcgccggcGACGTC<br>
&nbsp;&nbsp;&nbsp; CTTAAGcgccggcgaAGATCTacnnnnnnTGGCCAattaTGATCAtcgccggcGACGTC<br>
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R&nbsp; P&nbsp; L&nbsp; Q&nbsp;&nbsp; -</font></b></p>
R&nbsp; P&nbsp; L&nbsp; Q&nbsp;&nbsp; -</font></b></p>
<p><br></p>
<p><br></p>
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<h2>Discussion</h2>
<h2>Discussion</h2>
<h2>References</h2>
<h2>References</h2>

Revision as of 20:36, 21 October 2007

Contents

Proposal for a BioBrick extension for fusion proteins

iGEM team Freiburg

Introduction

Proposal

iGEM parts for mix-and-match construction of fusion proteins

proposed by Katja M. Arndt, Kristian M. Müller, University of Freiburg

  • Both parts are compatible with all standard iGEM parts as they have the BioBricks prefix for coding sequences and the standard BioBrick suffix.
  • nnnnnn is a place holder for the coding sequence of the respective part.
  • Both parts have two additional enzymes, NgoMIV and AgeI, which have compatible cohesive ends and enable in-frame fusion of protein parts with the linker sequence TG (no stop codons).
  • The only difference of the N-part and FusionPart is the additional NgoMIV site in the FusionPart.
  • The N-part is designed to be the start of a fusion protein or a stand-alone protein part, to be cloned via XbaI/PstI to any iGEM RBS expression part.
  • The FusionPart is designed to be fused to the N-part by digesting the N-part with AgeI/SpeI and the FusionPart with NgoMIV/SpeI.
  • Any number of FusionParts can be combined and fused to the N-part.
  • The FusionPart can also be a stand-alone part as it has a start codon after the XbaI site (BioBrick prefix for coding sequence), however two additional amino acids (A, G) are encoded before the start of the protein.

N-part

                             BsrFI                    SfcI
 ApoI                        BsaWI                 MspA1I|
EcoRI   NotI    XbaI          AgeI      SpeI    NotI    ||PstI
    |      |       |             |         |       |    ||   |
    GAATTCgcggccgctTCTAGAtgnnnnnnACCGGTtaatACTAGTagcggccgCTGCAG
  1 ---------+---------+---------+---------+---------+--------- 59
    CTTAAGcgccggcgaAGATCTacnnnnnnTGGCCAattaTGATCAtcgccggcGACGTC
c     I  R  G  R  F  *  M  ?  ?  T  G  *  Y  *  *  R  P  L  Q   -


FusionPart

                          NaeI     BsrFI                    SfcI
 ApoI                  BsrFI |     BsaWI                 MspA1I|
EcoRI   NotI    XbaI  NgoMIV |      AgeI      SpeI    NotI    ||PstI
    |      |       |       | |         |         |       |    ||   |
    GAATTCgcggccgctTCTAGAtgGCCGGCnnnnnnACCGGTtaatACTAGTagcggccgCTGCAG
  1 ---------+---------+---------+---------+---------+---------+----- 65
    CTTAAGcgccggcgaAGATCTacCGGCCGnnnnnnTGGCCAattaTGATCAtcgccggcGACGTC
c     I  R  G  R  F  *  M  A  G  ?  ?  T  G  *  Y  *  *  R  P  L  Q   -


Discussion

References