Wiki/Team:Warsaw/primers

From 2008.igem.org

(Difference between revisions)
Line 62: Line 62:
</pre>
</pre>
-
<h4>Fusion of the A protein with thwe OmpA
+
<h4>Fusion of the A protein with the Alpha part of beta-lactamase</h4>
-
 
+
<pre>
 +
<a name="AL+link10+homo2">AL+link10+homo2</a>  5' TCTGGAGGTGGAGGTAGCGG GGGTGGGGGTGGTTCGGGTGGAGGTGGT AAAACCGCGGCTCTTGCGC  3'
 +
<a name="AlphaP+link10+homo2">AlphaP+link10+homo2</a>  5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC TTCGCCAGTTAATAGTTTGCG  3'
 +
</pre>
<br><br><br><br>
<br><br><br><br>
Line 89: Line 92:
<a name="AL+SacI">AL+SacI</a>  5' GG GAGCTC T AAAACCGCGGCTCTTGCGCAAC  3'
<a name="AL+SacI">AL+SacI</a>  5' GG GAGCTC T AAAACCGCGGCTCTTGCGCAAC  3'
<a name="AP+NotI">AP+NotI</a>  5' AA GCGGCCGC C GGTTGACTTCCCCGCGGAATTC  3'
<a name="AP+NotI">AP+NotI</a>  5' AA GCGGCCGC C GGTTGACTTCCCCGCGGAATTC  3'
-
<a name="AL+link10+homo2">AL+link10+homo2</a>  5' TCTGGAGGTGGAGGTAGCGG GGGTGGGGGTGGTTCGGGTGGAGGTGGT AAAACCGCGGCTCTTGCGC  3'
+
 
<a name="AlphaL+SacI">AlphaL+SacI</a>  5' GG GAGCTC T CACCCAGAAACGCTGGTGAAAG  3'
<a name="AlphaL+SacI">AlphaL+SacI</a>  5' GG GAGCTC T CACCCAGAAACGCTGGTGAAAG  3'
-
<a name="AlphaP+link10+homo2">AlphaP+link10+homo2</a>  5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC TTCGCCAGTTAATAGTTTGCG  3'
+
 
<a name="OmegaL+SacI">OmegaL+SacI</a>  5' GG GAGCTC G CTACTTACTCTAGCTTCCCGG  3'
<a name="OmegaL+SacI">OmegaL+SacI</a>  5' GG GAGCTC G CTACTTACTCTAGCTTCCCGG  3'
<a name="OmegaP+link10+homo2">OmegaP+link10+homo2</a>  5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC CCAATGCTTAATCAGTGAGGC  3'
<a name="OmegaP+link10+homo2">OmegaP+link10+homo2</a>  5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC CCAATGCTTAATCAGTGAGGC  3'

Revision as of 15:37, 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

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'

Fusion of the A protein with the Alpha part of beta-lactamase

 
AL+link10+homo2   5' TCTGGAGGTGGAGGTAGCGG GGGTGGGGGTGGTTCGGGTGGAGGTGGT AAAACCGCGGCTCTTGCGC  3'
AlphaP+link10+homo2   5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC TTCGCCAGTTAATAGTTTGCG  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 ;) 


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'
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'
   5'   3'
   5'   3'
   5'   3'
   5'   3'
   5'   3'

Retrieved from "http://2008.igem.org/Wiki/Team:Warsaw/primers"