Wiki/Team:Warsaw/vectors/pACYC177+ompA-alpha-A-omega

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<h2>pACYC177_lac + ompA-alpha-A-omega</h2>
<h2>pACYC177_lac + ompA-alpha-A-omega</h2>
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<p class=vector>Description</p><br>
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<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-A-omega>pACYC177+ompA-A-omega</a><br>
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<b>Resistance</b>: kanamycin<br>
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<b>Features</b>: Fusion of OmpA, omega and alpha fragments of &beta;-lactamase and protein A under control of IPTG inducible lactose promoter. Low (10) copy plasmid<br>
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<b>Preparation</b>: Alpha fragment of &beta;-lactamase amplified from <a href=http://www.fermentas.com/techinfo/nucleicacids/mappuc1819.htm>pUC19</a> plasmid using
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<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlphaL+SacI">AlphaL+SacI</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlphaPlinkSac">AlphaPlinkSac</a>
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primers, digested using SacI and cloned into <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-omega>pACYC177+OmpA-A-omega</a> <b><a href=https://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=9&arg0=13_August_2008&arg1=14_August_2008&arg2=18_August_2008&arg3=19_August_2008&arg4=20_August_2008&arg5=21_August_2008&arg6=22_August_2008&arg7=25_August_2008&arg8=26_August_2008&name=Cloning%20of%20alpha%20to%20pACYC177%20OmpA_A_omega%20and%20pACYC177%20OmpA_Z_omega>[Notebook entries]</a></b>
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<br>
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<b>Usage:</b> OmpA-alpha-A-omega protein is directed to bacterial outer membrane thanks to presence of OmpA protein.  This is one of our 'hunter' protein variants<br>
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<br><br>
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<p class=vector>Vector map</p><br><br>
<img src="https://static.igem.org/mediawiki/2008/5/5b/PACYClac%2BompA-alpha-A-omega.png" width=600>
<img src="https://static.igem.org/mediawiki/2008/5/5b/PACYClac%2BompA-alpha-A-omega.png" width=600>
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<br><br>
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vector sequence:
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<p class=vector>Vector sequence</p>
<pre>
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Latest revision as of 17:02, 28 October 2008

Gallery Bricks Notebook Team Project Home


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Vectors view

 

pACYC177_lac + ompA-alpha-A-omega

Description


Author: Michał K.
Based on: pACYC177+ompA-A-omega
Resistance: kanamycin
Features: Fusion of OmpA, omega and alpha fragments of β-lactamase and protein A under control of IPTG inducible lactose promoter. Low (10) copy plasmid
Preparation: Alpha fragment of β-lactamase amplified from pUC19 plasmid using AlphaL+SacI and AlphaPlinkSac primers, digested using SacI and cloned into pACYC177+OmpA-A-omega [Notebook entries]
Usage: OmpA-alpha-A-omega 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

CTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGG
GTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAAC
GTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTG
ACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGT
ACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTG
CTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGAC
CGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTT
GGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAG
CAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAAGGTGGCGGAGGAAGCGGAGGAG
GTGGTTCTGGAGGTGGAGGTAGCGGGGGTGGGGGTGGTTCGGGTGGAGGTGGTAAAACCG
CGGCTCTTGCGCAACACGATGAAGCCGTGGACAACAAATTCAACAAAGAACAACAAAACG
CGTTCTATGAGATCTTACATTTACCTAACTTAAACGAAGAACAACGAAACGCCTTCATCC
AAAGTTTAAAAGATGACCCAAGCCAAAGCGCTAACCTTTTAGCAGAAGCTAAAAAGCTAA
ATGATGCTCAGGCGCCGAAAGTAGACAACAAATTCAACAAAGAACAACAAAACGCGTTCT
ATGAGATCTTACATTTACCTAACTTAAACGAAGAACAACGAAACGCCTTCATCCAAAGTT
TAAAAGATGACCCAAGCCAAAGCGCTAACCTTTTAGCAGAAGCTAAAAAGCTAAATGGTG
CTCAGGCGCCGAAAGTAGACGCGAATTCCGCGGGGAAGTCAACCGGCGGCCGCAGTGGTG
GAAGCGGTGGCGGAGGATCTGGAGGAGGTGGTTCTGGAGGTGGAGGTCTACTTACTCTAG
CTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGC
GCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGT
CTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCT
ACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTG
CCTCACTGATTAAGCATTGGTAAGAATTCTTCTGCAGTTGGATCCTCCGGCGTTCAGCCT
GTGCCACAGCCGACAGGATGGTGACCACCATTTGCCCCATATCACCGTCGGTACTGATCC
CGTCGTCAATAAACCGAACCGCTACACCCTGAGCATCAAACTCTTTTATCAGTTGGATCA
TGTCGGCGGTGTCGCGGCCAAGACGGTCGAGCTTCTTCACCAGAATGACATCACCTTCCT
CCACCTTCATCCTCAGCAAATCCAGCCCTTCCCGATCTGTTGAACTGCCGGATGCCTTGT
CGGTAAAGATGCGGTTAGCTTTTACCCCTGCATCTTTGAGCGCTGAGGTCTGCCTCGTGA
AGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGCCAGAAAGTGAG
GGAGCCACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTTGGTGATTTTGAACTTTTG
CTTTGCCACGGAACGGTCTGCGTTGTCGGGAAGATGCGTGATCTGATCCTTCAACTCAGC
AAAAGTTCGATTTATTCAACAAAGCCGCCGTCCCGTCAAGTCAGCGTAATGCTCTGCCAG
TGTTACAACCAATTAACCAATTCTGATTAGAAAAACTCATCGAGCATCAAATGAAACTGC
AATTTATTCATATCAGGATTATCAATACCATATTTTTGAAAAAGCCGTTTCTGTAATGAA
GGAGAAAACTCACCGAGGCAGTTCCATAGGATGGCAAGATCCTGGTATCGGTCTGCGATT
CCGACTCGTCCAACATCAATACAACCTATTAATTTCCCCTCGTCAAAAATAAGGTTATCA
AGTGAGAAATCACCATGAGTGACGACTGAATCCGGTGAGAATGGCAAAAGCTTATGCATT
TCTTTCCAGACTTGTTCAACAGGCCAGCCATTACGCTCGTCATCAAAATCACTCGCATCA
ACCAAACCGTTATTCATTCGTGATTGCGCCTGAGCGAGACGAAATACGCGATCGCTGTTA
AAAGGACAATTACAAACAGGAATCGAATGCAACCGGCGCAGGAACACTGCCAGCGCATCA
ACAATATTTTCACCTGAATCAGGATATTCTTCTAATACCTGGAATGCTGTTTTCCCGGGG
ATCGCAGTGGTGAGTAACCATGCATCATCAGGAGTACGGATAAAATGCTTGATGGTCGGA
AGAGGCATAAATTCCGTCAGCCAGTTTAGTCTGACCATCTCATCTGTAACATCATTGGCA
ACGCTACCTTTGCCATGTTTCAGAAACAACTCTGGCGCATCGGGCTTCCCATACAATCGA
TAGATTGTCGCACCTGATTGCCCGACATTATCGCGAGCCCATTTATACCCATATAAATCA
GCATCCATGTTGGAATTTAATCGCGGCCTCGAGCAAGACGTTTCCCGTTGAATATGGCTC
ATAACACCCCTTGTATTACTGTTTATGTAAGCAGACAGTTTTATTGTTCATGATGATATA
TTTTTATCTTGTGCAATGTAACATCAGAGATTTTGAGACACAACGTGGCTTTGTTGAATA
AATCGAACTTTTGCTGAGTTGAAGGATCAGATCACGCATCTTCCCGACAACGCAGACCGT
TCCGTGGCAAAGCAAAAGTTCAAAATCACCAACTGGTCCACCTACAACAAAGCTCTCATC
AACCGTGGCTCCCTCACTTTCTGGCTGGATGATGGGGCGATTCAGGCCTGGTATGAGTCA
GCAACACCTTCTTCACGAGGCAGACCTCAGCGCTAGCGGAGTGTATACTGGCTTACTATG
TTGGCACTGATGAGGGTGTCAGTGAAGTGCTTCATGTGGCAGGAGAAAAAAGGCTGCACC
GGTGCGTCAGCAGAATATGTGATACAGGATATATTCCGCTTCCTCGCTCACTGACTCGCT
ACGCTCGGTCGTTCGACTGCGGCGAGCGGAAATGGCTTACGAACGGGGCGGAGATTTCCT
GGAAGATGCCAGGAAGATACTTAACAGGGAAGTGAGAGGGCCGCGGCAAAGCCGTTTTTC
CATAGGCTCCGCCCCCCTGACAAGCATCACGAAATCTGACGCTCAAATCAGTGGTGGCGA
AACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGCGGCTCCCTCGTGCGCTCT
CCTGTTCCTGCCTTTCGGTTTACCGGTGTCATTCCGCTGTTATGGCCGCGTTTGTCTCAT
TCCACGCCTGACACTCAGTTCCGGGTAGGCAGTTCGCTCCAAGCTGGACTGTATGCACGA
ACCCCCCGTTCAGTCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCC
GGAAAGACATGCAAAAGCACCACTGGCAGCAGCCACTGGTAATTGATTTAGAGGAGTTAG
TCTTGAAGTCATGCGCCGGTTAAGGCTAAACTGAAAGGACAAGTTTTGGTGACTGCGCTC
CTCCAAGCCAGTTACCTCGGTTCAAAGAGTTGGTAGCTCAGAGAACCTTCGAAAAACCGC
CCTGCAAGGCGGTTTTTTCGTTTTCAGAGCAAGAGATTACGCGCAGACCAAAACGATCTC
AAGAAGATCATCTTATTAAGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGA
TTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAA
GTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAA
TCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCC
CCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGA
TACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAA
GGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTT
GCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTG
CTGCAGCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGC
ACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGC
TCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAA
TTGTGAGCGGATAACAATTTCACACAGGAAACACATATGAAAGCTACTAAACTGGTACTG
GGCAACAACAATGGCCCGACCCATGAAAACCAACTGGGCGCTGGTGCTTTTGGTGGTTAC
CAGGTTAACCCGTATGTTGGCTTTGAAATGGGTTACGACTGGTTAGGTCGTATGCCGTAC
AAAGGCAGCGTTGAAAACGGTGCATACAAAGCTCAGGGCGTTCAACTGACCGCTAAACTG
GGTTACCCAATCACTGACGACCTGGACATCTACACTCGTCTGGGTGGCATGGTATGGCGT
GCAGACACTAAATCCAACGTTTATGGTAAAAACCACGACACCGGCGTTTCTCCGGTCTTC
GCTGGCGGTGTTGAGTACGCGATCACTCCTGAAATCGCTACCCGTCTGGAATACCAGTGG
ACCAACAACATCGGTGACGCACACACCATCGGCACTCGTCCGGACAACGGCGGAGGTTCT
GGAGGAGGGAGCT