Freiburg07/report fusion parts
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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> | ||
</ul> | </ul> | ||
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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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| - | <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> | + | <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> |
1 ---------+---------+---------+---------+---------+--------- 59<br> | 1 ---------+---------+---------+---------+---------+--------- 59<br> | ||
CTTAAGcgccggcgaAGATCTacnnnnnnTGGCCAattaTGATCAtcgccggcGACGTC<br> | CTTAAGcgccggcgaAGATCTacnnnnnnTGGCCAattaTGATCAtcgccggcGACGTC<br> | ||
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R P L Q -</font></b></p> | R P L Q -</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 -
