Wiki/Team:Warsaw/vectors/pACYC177+ompA-deltaA-alpha
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<p class=vector>Description</p><br> | <p class=vector>Description</p><br> | ||
<b>Author</b>: Michał K.<br> | <b>Author</b>: Michał K.<br> | ||
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<b>Based on</b>: <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-alpha>pACYC177+OmpA-alpha</a><br> | <b>Based on</b>: <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-alpha>pACYC177+OmpA-alpha</a><br> | ||
+ | <b>Resistance</b>: kanamycin<br> | ||
+ | <b>Features</b>: Fusion of OmpA, alpha fragment of β-lactamase and B-domain of protein A (ΔA) under control of IPTG inducible lactose promoter. Low (10) copy plasmid<br> | ||
<b>Preparation</b>: ΔA coding sequence amplified from pDRIVE-TapTag using <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AL+SacI ">AL+SacI</a> and | <b>Preparation</b>: ΔA coding sequence amplified from pDRIVE-TapTag using <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AL+SacI ">AL+SacI</a> and | ||
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AP+NotI">AP+NotI</a> <br> | <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AP+NotI">AP+NotI</a> <br> | ||
- | primers. Resulting fragment was digested with NotI and SacI and ligated into <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-alpha>pACYC177+OmpA-alpha</a> | + | primers. Resulting fragment was digested with NotI and SacI and ligated into <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-alpha>pACYC177+OmpA-alpha</a> <b><a href=https://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=5&arg0=30_July_2008&arg1=31_July_2008&arg2=1_August_2008&arg3=4_August_2008&arg4=5_August_2008&name=Cloning%20of%20truncated%20fragment%20of%20protein%20A%20(%26Delta%3BA)>[Notebook entries]</a></b> |
<br> | <br> | ||
<b>Usage:</b> OmpA-ΔA-alpha protein is directed to bacterial outer membrane thanks to presence of OmpA protein. This is one of our 'hunter' protein variants<br> | <b>Usage:</b> OmpA-ΔA-alpha protein is directed to bacterial outer membrane thanks to presence of OmpA protein. This is one of our 'hunter' protein variants<br> |
Latest revision as of 17:47, 28 October 2008
pACYC177_lac + ompA-ΔA-alphaDescription Author: Michał K. Based on: pACYC177+OmpA-alpha Resistance: kanamycin Features: Fusion of OmpA, alpha fragment of β-lactamase and B-domain of protein A (ΔA) under control of IPTG inducible lactose promoter. Low (10) copy plasmid Preparation: ΔA coding sequence amplified from pDRIVE-TapTag using AL+SacI and AP+NotI primers. Resulting fragment was digested with NotI and SacI and ligated into pACYC177+OmpA-alpha [Notebook entries] Usage: OmpA-ΔA-alpha protein is directed to bacterial outer membrane thanks to presence of OmpA protein. This is one of our 'hunter' protein variants Vector map Vector sequence TATGTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGA AGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTG CGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGC CAACGCGCGGGGAGAGGCGGTTTGCGTATTGCTGCAGCAATGGCAACAACGTTGCGCAAA CTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAG GCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCT GATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGAT GGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAA CGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGAC CAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATC TAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTC CACTGAGCGTCAGACCCCTTAATAAGATGATCTTCTTGAGATCGTTTTGGTCTGCGCGTA ATCTCTTGCTCTGAAAACGAAAAAACCGCCTTGCAGGGCGGTTTTTCGAAGGTTCTCTGA GCTACCAACTCTTTGAACCGAGGTAACTGGCTTGGAGGAGCGCAGTCACCAAAACTTGTC CTTTCAGTTTAGCCTTAACCGGCGCATGACTTCAAGACTAACTCCTCTAAATCAATTACC AGTGGCTGCTGCCAGTGGTGCTTTTGCATGTCTTTCCGGGTTGGACTCAAGACGATAGTT ACCGGATAAGGCGCAGCGGTCGGACTGAACGGGGGGTTCGTGCATACAGTCCAGCTTGGA GCGAACTGCCTACCCGGAACTGAGTGTCAGGCGTGGAATGAGACAAACGCGGCCATAACA GCGGAATGACACCGGTAAACCGAAAGGCAGGAACAGGAGAGCGCACGAGGGAGCCGCCAG GGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCACTGATTTGAGCGTC AGATTTCGTGATGCTTGTCAGGGGGGCGGAGCCTATGGAAAAACGGCTTTGCCGCGGCCC TCTCACTTCCCTGTTAAGTATCTTCCTGGCATCTTCCAGGAAATCTCCGCCCCGTTCGTA AGCCATTTCCGCTCGCCGCAGTCGAACGACCGAGCGTAGCGAGTCAGTGAGCGAGGAAGC GGAATATATCCTGTATCACATATTCTGCTGACGCACCGGTGCAGCCTTTTTTCTCCTGCC ACATGAAGCACTTCACTGACACCCTCATCAGTGCCAACATAGTAAGCCAGTATACACTCC GCTAGCGCTGAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCG CCCCATCATCCAGCCAGAAAGTGAGGGAGCCACGGTTGATGAGAGCTTTGTTGTAGGTGG ACCAGTTGGTGATTTTGAACTTTTGCTTTGCCACGGAACGGTCTGCGTTGTCGGGAAGAT GCGTGATCTGATCCTTCAACTCAGCAAAAGTTCGATTTATTCAACAAAGCCACGTTGTGT CTCAAAATCTCTGATGTTACATTGCACAAGATAAAAATATATCATCATGAACAATAAAAC TGTCTGCTTACATAAACAGTAATACAAGGGGTGTTATGAGCCATATTCAACGGGAAACGT CTTGCTCGAGGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGG CTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGATTGTATGGGAAGCCCGATG CGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGA TGGTCAGACTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCC GTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGGAAAACAGCATTCCAGG TATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGC GCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGTC TCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGATGCGAGTGATTTTGATGACG AGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAGCTTTTGCCATTCT CACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGG GGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATC TTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTC AAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATG AGTTTTTCTAATCAGAATTGGTTAATTGGTTGTAACACTGGCAGAGCATTACGCTGACTT GACGGGACGGCGGCTTTGTTGAATAAATCGAACTTTTGCTGAGTTGAAGGATCAGATCAC GCATCTTCCCGACAACGCAGACCGTTCCGTGGCAAAGCAAAAGTTCAAAATCACCAACTG GTCCACCTACAACAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGATGATGG GGCGATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCTC AAAGATGCAGGGGTAAAAGCTAACCGCATCTTTACCGACAAGGCATCCGGCAGTTCAACA GATCGGGAAGGGCTGGATTTGCTGAGGATGAAGGTGGAGGAAGGTGATGTCATTCTGGTG AAGAAGCTCGACCGTCTTGGCCGCGACACCGCCGACATGATCCAACTGATAAAAGAGTTT GATGCTCAGGGTGTAGCGGTTCGGTTTATTGACGACGGGATCAGTACCGACGGTGATATG GGGCAAATGGTGGTCACCATCCTGTCGGCTGTGGCACAGGCTGAACGCCGGAGGATCCGG TCTAGATTATTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGT GGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCG AGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGT TGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTC TCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTC ATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAA TACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCG AAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACC CAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGACCTCCACCTCCAGA ACCACCTCCTCCAGATCCTCCGCCACCGCTTCCACCACTGCGGCCGCCGGTTGACTTCCC CGCGGAATTCGCGTCTACTTTCGGCGCCTGAGCATCATTTAGCTTTTTAGCTTCTGCTAA AAGGTTAGCGCTTTGGCTTGGGTCATCTTTTAAACTTTGGATGAAGGCGTTTCGTTGTTC TTCGTTTAAGTTAGGTAAATGTAAGATCTCATAGAACGCGTTTTGTTGTTCTTTGTTGAA TTTGTTGTCCACGGCTTCATCGTGTTGCGCAAGAGCCGCGGTTTTAGAGCTCCCTCCTCC AGAACCTCCGCCGTTGTCCGGACGAGTGCCGATGGTGTGTGCGTCACCGATGTTGTTGGT CCACTGGTATTCCAGACGGGTAGCGATTTCAGGAGTGATCGCGTACTCAACACCGCCAGC GAAGACCGGAGAAACGCCGGTGTCGTGGTTTTTACCATAAACGTTGGATTTAGTGTCTGC ACGCCATACCATGCCACCCAGACGAGTGTAGATGTCCAGGTCGTCAGTGATTGGGTAACC CAGTTTAGCGGTCAGTTGAACGCCCTGAGCTTTGTATGCACCGTTTTCAACGCTGCCTTT GTACGGCATACGACCTAACCAGTCGTAACCCATTTCAAAGCCAACATACGGGTTAACCTG GTAACCACCAAAAGCACCAGCGCCCAGTTGGTTTTCATGGGTCGGGCCATTGTTGTTGCC CAGTACCAGTTTAGTAGCTTTCA |