Wiki/Team:Warsaw/primers

From 2008.igem.org

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<h4>Mutagenesis of the A protein</h4>
<h4>Mutagenesis of the A protein</h4>
<pre>
<pre>
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<a name="AmutDelecjaL+KpnI">AmutDelecjaL+KpnI</a> 5' AGGTACCCATGTAAGATCTCATAGAACGCG  3'
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<a name="ADelL+KpnI">ADe;L+KpnI</a> 5' AGGTACCCATGTAAGATCTCATAGAACGCG  3'
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<a name="AmutDelecjaP">AmutDelecjaP</a>      5' AACTTAAACGAAGAACAACGAAACGCC  3'
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<a name="ADelP">AdelP</a>      5' AACTTAAACGAAGAACAACGAAACGCC  3'
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<a name="AmutWiecejL+KpnI">AmutWiecejL+KpnI</a>  5' TCGTTTAAGTTAGGTACCGGTGAGATCTCAGAGGACGCGTTTTGTTGTTCTTTGTTG  3'
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<a name="AmutL+KpnI">AmutL+KpnI</a>  5' TCGTTTAAGTTAGGTACCGGTGAGATCTCAGAGGACGCGTTTTGTTGTTCTTTGTTG  3'
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<a name="AmutWiecejP">AmutWiecejP</a>      5' AGAACAAGGAAACGCCTTCGCCCAAAGTTTAGCAGATGACCCAAGCCAAAGCGC  3'
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<a name="AmutpP">AmutP</a>      5' AGAACAAGGAAACGCCTTCGCCCAAAGTTTAGCAGATGACCCAAGCCAAAGCGC  3'
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<a name="AlphaPlinkSac">AlphaPlinkSac</a>      5' TTGAGCTCCCTCCACCTCCGCTACCTCCACCAGAACCACCTCCTCCGCTTCCGGTACCTTCGCCAGTTAATAGTTTGC  3'
<a name="AlphaPlinkSac">AlphaPlinkSac</a>      5' TTGAGCTCCCTCCACCTCCGCTACCTCCACCAGAACCACCTCCTCCGCTTCCGGTACCTTCGCCAGTTAATAGTTTGC  3'
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<h2>BioBricks Preparation</h2>
<h2>BioBricks Preparation</h2>
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<a name="AP+XhoI+SpeI">AP+XhoI+SpeI</a>        5' AA ACTAGT CTCGAG GGTTGACTTCCCCGCGGAATTC  3'
<a name="AP+XhoI+SpeI">AP+XhoI+SpeI</a>        5' AA ACTAGT CTCGAG GGTTGACTTCCCCGCGGAATTC  3'
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Revision as of 15:52, 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'

Sequencing of the T7 polymerase gene

T7Rseq      5' GCTTCAACCGCAGCTACCAC 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

ADe;L+KpnI 5' AGGTACCCATGTAAGATCTCATAGAACGCG  3'
AdelP      5' AACTTAAACGAAGAACAACGAAACGCC  3'
AmutL+KpnI  5' TCGTTTAAGTTAGGTACCGGTGAGATCTCAGAGGACGCGTTTTGTTGTTCTTTGTTG  3'
AmutP       5' AGAACAAGGAAACGCCTTCGCCCAAAGTTTAGCAGATGACCCAAGCCAAAGCGC  3'

Fusion of the A protein with the OmpaA protein and the alpha/omega part of beta-lactamase protein

 
AL+link10+homo2     5' TCTGGAGGTGGAGGTAGCGG GGGTGGGGGTGGTTCGGGTGGAGGTGGT AAAACCGCGGCTCTTGCGC  3'
AlphaP+link10+homo2 5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC TTCGCCAGTTAATAGTTTGCG  3'
OmegaP+link10+homo2 5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC CCAATGCTTAATCAGTGAGGC  3'
AL+SacI             5' GG GAGCTC T AAAACCGCGGCTCTTGCGCAAC  3'
AP+NotI             5' AA GCGGCCGC C GGTTGACTTCCCCGCGGAATTC  3'
AlphaL+SacI         5' GG GAGCTC T CACCCAGAAACGCTGGTGAAAG  3'
OmegaL+SacI         5' GG GAGCTC G CTACTTACTCTAGCTTCCCGG  3'
AlphaPlinkSac       5' TTGAGCTCCCTCCACCTCCGCTACCTCCACCAGAACCACCTCCTCCGCTTCCGGTACCTTCGCCAGTTAATAGTTTGC  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'
AP+XhoI+SpeI         5' AA ACTAGT CTCGAG GGTTGACTTCCCCGCGGAATTC  3'


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