Wiki/Team:Warsaw/primers

From 2008.igem.org

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<h2>BioBricks Preparation</h2>
<h2>BioBricks Preparation</h2>
<pre>
<pre>
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<a name=""></a>  5'   3'
+
<a name="AIDpHSNP">AIDP_HindSpeNotPst</a>  5' AAACTGCAGCGGCCGCTACTAGTAAAGCTTTCAAAGTCCCAAAGTACGAAATG  3'
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<a name=""></a>  5'   3'
+
<a name="AL_BNXNE">AL_BNXNE</a>  5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGGGATCCAAAACCGCGGCTCTTGCGCAAC  3'
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<a name=""></a>  5'   3'
+
<a name="AlphaPSpe">AlfaPSpe</a>  5' GCACTAGTATTATTCGCCAGTTAATAGTTTGCG  3'
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<a name=""></a>  5'   3'
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<a name="APSacSpe">APSacSpe</a>  5' GGACTAGTAGAGCTCCCGTCTACTTTCGGCGCCTGAGC  3'
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<a name=""></a>  5'   3'
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<a name="LinLSXNE">LinLSXNE</a>  5' ATGAATTCGCGGCCGCTTCTAGAGGGAGCTCTGGTGGAAGCGGTGGCGGAGGATC  3'
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<a name=""></a>  5'   3'
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<a name="LinP_BS">LinP_BS</a>  5' CCACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTAC  3'
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<a name=""></a>  5'   3'
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<a name="LinPBSNP">LinPBSNP</a>  5' AGCCTGCAGCGGCCGCTACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTACC  3'
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<a name=""></a>  5'   3'
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<a name="OmegPSpe">OmegPSpe</a>  5' GCACTAGTATTACCAATGCTTAATCAGTGAG  3'
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<a name=""></a>  5'   3'
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<a name="OmegLNde">OmegLNde</a>  5' CAGCCATATGCTACTTACTCTAGCTTCCCGG  3'
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<a name=""></a>  5'   3'
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<a name="OmpLNXNE">OmpLNXNE</a>  5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTAC  3'
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<a name=""></a>  5'   3'
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<a name="OmLNXNEB">OmLNXNEB</a>  5' ATTAGATCTGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTAC  3'
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<a name=""></a>  5'  3'
+
<a name="PETt7L_XNE">pETt7L_XNE</a>  5' ATGAATTCGCGGCCGCTTCTAGAGATCCCGCGAAATTAATACG  3'
 +
<a name="PlacL_XNE">PlacL_XNE</a>   5' TAGAATTCGCGGCCGCTTCTAGAGCTGGCACGACAGGTTTCCC  3'
</pre>
</pre>

Revision as of 15:09, 26 September 2008

Gallery Bricks Notebook Team Project Home

 

Primers

One-armed bandit preparation

AID and T7 cloning

T7lLinkB   5' CGGATCCGGCGGTGGCGGTAGTGGAGGAGGTGGGTCTGGAGGCGGAGGGAACACGATTAACATCGCTAAGAAC  3'
T7lRBSHi   5' CACAAGCTTAGGAGGTGTACTAGATGAACACGATTAACATCGCTAAG  3'
T7pXbSal   5' TCTGTCGACTCTAGATTACGCGAACGCGAAGTCC  3'
AIDlNrH    5' CGTAAGCTTAGGAGGTGTAGACCATGGACAGCCTCTTGATG  3'
AIDpLinB   5' CCACTACCGCCACCGCCGGATCCGCCACCTCCGCTGCCTCCACCACCAAGTCCCAAAGTACGAAATGC  3'

Sequencing of pMPM-T5

pMPMT5seqF  5' CTACTGTTTCTCCATACCCG  3'
pMPMT5seqR  5' GAAAATCTTCTCTCATCCGC 3'
AIDseqP     5' GCTTCGGCGCATCCTTTTG 3'

Sequencing of β-galactosidase from pZC320

pZCseqL     5' CTTGACCCGCAGTTGCAAAC 3'
pZCseqP     5' CCCAACTGATCTTCAGCATC 3'

Hunter and prey

Fusion of protein A and ompA and alpha or omega part of beta-lactamase

alphaL+link14+homo (alphaL_link)          5' CAGTGGTGGAAGCGGTGGC GGAGGATCTGGAGGAGGTGGTTCTGGAGGTGGAGGT CACCCAGAAACGCTGGTGAAAG 3'
alphaP+STOP+XbaI+BamHI (alphaP_XB)        5' GG GGATCC GG TCTAGA TTA TTCGCCAGTTAATAGTTTGCG 3'
omegaL+link14+homo (omegaL_link)          5' CAGTGGTGGAAGCGGTGGC GGAGGATCTGGAGGAGGTGGTTCTGGAGGTGGAGGT CTACTTACTCTAGCTTCCCGG 3'
omegaP+STOP+EcoRI+PstI+BamHI (omegaP_EPB) 5' AAGGATCCAACTGCAGAAGAATTC TTACCAATGCTTAATCAGTG 3'
OmpAL+NdeI (ompaL_N)                      5' CATATGAAAGCTACTAAACTGGTACTGGG 3'
OmpAP+link8+SacI+NotI+homo (ompaP_link)   5' GCCACCGCTTCCACCACT GCGGCCGC CA GAGCTC CCTCCTCCAGAACCTCC GCCGTTGTCCGGACGAGTGCCGATG 3'

Cloning of A and/or Z protein to expression vector (pET15b/pET24a)

AlphaP+link+Sac   5' GG GAGCTC CC ACCACCCCCACCCCCGCTACCTC  3'
AlphaL+Nde        5' CGGG CATATG CACCCAGAAACGCTGGTGAAAG  3'

Mutagenesis of the A protein

AmutDelecjaL+KpnI   5' AGGTACCCATGTAAGATCTCATAGAACGCG  3'
AmutDelecjaP   5' AACTTAAACGAAGAACAACGAAACGCC  3'
AmutMniejL+KpnI   5' AGTTAGGTACCGGTGAGATCTCATAGGACGCGTTTTGTTGTTCTTTGTTG  3'
AmutMniejP   5' TAAACGAAGAACAAGGAAACGCCTTCATCCAAAGTTTAAAAGATGACCC  3'
AmutWiecejL+KpnI   5' TCGTTTAAGTTAGGTACCGGTGAGATCTCAGAGGACGCGTTTTGTTGTTCTTTGTTG  3'
AmutWiecejP   5' AGAACAAGGAAACGCCTTCGCCCAAAGTTTAGCAGATGACCCAAGCCAAAGCGC  3'




BioBricks Preparation

AIDP_HindSpeNotPst   5' AAACTGCAGCGGCCGCTACTAGTAAAGCTTTCAAAGTCCCAAAGTACGAAATG  3'
AL_BNXNE   5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGGGATCCAAAACCGCGGCTCTTGCGCAAC  3'
AlfaPSpe   5' GCACTAGTATTATTCGCCAGTTAATAGTTTGCG  3'
APSacSpe   5' GGACTAGTAGAGCTCCCGTCTACTTTCGGCGCCTGAGC  3'
LinLSXNE   5' ATGAATTCGCGGCCGCTTCTAGAGGGAGCTCTGGTGGAAGCGGTGGCGGAGGATC  3'
LinP_BS   5' CCACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTAC  3'
LinPBSNP   5' AGCCTGCAGCGGCCGCTACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTACC  3'
OmegPSpe   5' GCACTAGTATTACCAATGCTTAATCAGTGAG  3'
OmegLNde   5' CAGCCATATGCTACTTACTCTAGCTTCCCGG  3'
OmpLNXNE   5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTAC  3'
OmLNXNEB   5' ATTAGATCTGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTAC  3'
pETt7L_XNE   5' ATGAATTCGCGGCCGCTTCTAGAGATCCCGCGAAATTAATACG  3'
PlacL_XNE   5' TAGAATTCGCGGCCGCTTCTAGAGCTGGCACGACAGGTTTCCC  3'

For now - unclassified ;)

pZCt7l   5' TACCTTAGGTATTACGCGCGCTTGG  3'
pZCt7p   5' ATCCTAAGGTATAGGGCGGCCGCTAG  3'
AL+SacI   5' GG GAGCTC T AAAACCGCGGCTCTTGCGCAAC  3'
AP+NotI   5' AA GCGGCCGC C GGTTGACTTCCCCGCGGAATTC  3'
AL+link10+homo2   5' TCTGGAGGTGGAGGTAGCGG GGGTGGGGGTGGTTCGGGTGGAGGTGGT AAAACCGCGGCTCTTGCGC  3'
AlphaL+SacI   5' GG GAGCTC T CACCCAGAAACGCTGGTGAAAG  3'
AlphaP+link10+homo2   5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC TTCGCCAGTTAATAGTTTGCG  3'
OmegaL+SacI   5' GG GAGCTC G CTACTTACTCTAGCTTCCCGG  3'
OmegaP+link10+homo2   5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC CCAATGCTTAATCAGTGAGGC  3'
AlphaP+link+SacI   5' GG GAGCTC CC ACCACCCCCACCCCCGCTACCTC  3'
AlphaL+NdeI        5' CGGG CATATG CACCCAGAAACGCTGGTGAAAG  3'
AP+XhoI+SpeI   5' AA ACTAGT CTCGAG GGTTGACTTCCCCGCGGAATTC  3'
AL+NdeI+XbaI   5' GG TCTAGA AA CATATG AAAACCGCGGCTCTTGCGCAAC  3'
AlpaLNde   5' CCCATATGCACCCAGAAACGCTGGTGAAAG  3'
AlphaPlinkSac   5' TTGAGCTCCCTCCACCTCCGCTACCTCCACCAGAACCACCTCCTCCGCTTCCGGTACCTTCGCCAGTTAATAGTTTGC  3'
   5'   3'
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