Wiki/Team:Warsaw/primers

From 2008.igem.org

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<h2>Preparation of the BioBricks Standard Parts</h2>
<h2>Preparation of the BioBricks Standard Parts</h2>
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<a name="AIDpHSNP">AIDP_HindSpeNotPst</a>   5' AAACTGCAGCGGCCGCTACTAGTAAAGCTTTCAAAGTCCCAAAGTACGAAATG 3'
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<a name="AIDpSNP">AIDP_SpeNotPst</a>       5' AAACTGCAGCGGCCGCTACTAGTATCAAAGTCCCAAAGTACGAAATG 3'
<a name="AL_BNXNE">AL_BNXNE</a>            5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGGGATCCAAAACCGCGGCTCTTGCGCAAC  3'
<a name="AL_BNXNE">AL_BNXNE</a>            5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGGGATCCAAAACCGCGGCTCTTGCGCAAC  3'
<a name="AlphaPSpe">AlphaPSpe</a>            5' GCACTAGTATTATTCGCCAGTTAATAGTTTGCG  3'
<a name="AlphaPSpe">AlphaPSpe</a>            5' GCACTAGTATTATTCGCCAGTTAATAGTTTGCG  3'

Revision as of 14:23, 28 October 2008

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Primers

'One-armed bandit' strain 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'
pZCseqR     5' CCCAACTGATCTTCAGCATC 3'

Sequencing of the T7 polymerase gene

T7-2seq   5' AGGTGCGCTGGCTGGCATC 3'
T7-3seq   5' AAGGAAGGTTACTACTGGCT 3'
T7-4seq   5' CAAAGAGTTCGGCTTCCGTC 3'
T7Rseq    5' GCTTCAACCGCAGCTACCAC 3'

'Hunter and prey' strain preparation

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

ADelL+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'

Preparation of the BioBricks Standard Parts

AIDP_SpeNotPst        5' AAACTGCAGCGGCCGCTACTAGTATCAAAGTCCCAAAGTACGAAATG  3'
AL_BNXNE             5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGGGATCCAAAACCGCGGCTCTTGCGCAAC  3'
AlphaPSpe             5' GCACTAGTATTATTCGCCAGTTAATAGTTTGCG  3'
APSacSpe             5' GGACTAGTAGAGCTCCCGTCTACTTTCGGCGCCTGAGC  3'
LinLSXNE             5' ATGAATTCGCGGCCGCTTCTAGAGGGAGCTCTGGTGGAAGCGGTGGCGGAGGATC  3'
LinP_BS              5' CCACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTAC  3'
LinP_BS2             5' ATACTAGTACCGGATCCAGAACCTCCTCCACCGCTACCTCCACCTCCAGAAC  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+0XhoI+SpeI        5' AA ACTAGT CTCGAG GGTTGACTTCCCCGCGGAATTC  3'
AIDl                 5' TATCTAGAACCATGGACAGCCTCTTG 3'
AraCl                5' GCTCTAGATTATGACAACTTGACGGC 3'


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