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

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<h2>pACYC177_lac + ompA-alpha</h2>
<h2>pACYC177_lac + ompA-alpha</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>pACYC177_plac</a><br>
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<b>Resistance</b>: kanamycin<br>
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<b>Features</b>: Fusion of OmpA protein and alpha fragment of &beta;-lactamase under control of IPTG inducible lactose promoter. Low (10) copy plasmid<br>
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<b>Preparation</b>: OmpA fragment amplified from <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pB30D>pB30D</a> plasmid with
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<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaL_N">OmpaL_N</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaP_link">OmpaP_link</a>
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primers. Alpha fragment of &beta;-lactamase amplified from <a href=http://www.fermentas.com/techinfo/nucleicacids/mappuc1819.htm>pUC19</a> plasmid with <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlphaL_link">AlphaL_link</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlphaP_XB">AlphaP_XB</a> primers. Resulting fragments joined by PCR using <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaL_N">OmpaL_N</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlphaP_XB">AlphaP_XB</a> primers. Resulting fragment cutted with NdeI and BamHI and ligated into <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177>pACYC177_plac</a> <b>[<a href=https://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=11&arg0=13_June_2008&arg1=16_June_2008&arg2=17_June_2008&arg3=18_June_2008&arg4=19_June_2008&arg5=20_June_2008&arg6=23_June_2008&arg7=24_June_2008&arg8=25_June_2008&arg9=26_June_2008&arg10=27_June_2008&name=Preparation%20of%20constructs%3A%20OmpA_alpha%20and%20OmpA_omega>notebook entries</a> and <a href=https://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=6&arg0=7_July_2008&arg1=8_July_2008&arg2=9_July_2008&arg3=10_July_2008&arg4=11_July_2008&arg5=12_July_2008&name=Preparation%20of%20constructs%3A%20OmpA_alpha%20and%20OmpA_omega%20%232>another</a>]</b<br>
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<b>Usage:</b> Negative control in all of 'hunter and prey' tests <br>
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<br><br>
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<p class=vector>Vector map</p>
<img src="https://static.igem.org/mediawiki/2008/f/f8/PACYClac%2BompA-alpha.png" width=600>
<img src="https://static.igem.org/mediawiki/2008/f/f8/PACYClac%2BompA-alpha.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>
<pre>

Latest revision as of 16:54, 28 October 2008

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Vectors view

 

pACYC177_lac + ompA-alpha

Description


Author: Michał K.
Based on: pACYC177_plac
Resistance: kanamycin
Features: Fusion of OmpA protein and alpha fragment of β-lactamase under control of IPTG inducible lactose promoter. Low (10) copy plasmid
Preparation: OmpA fragment amplified from pB30D plasmid with OmpaL_N and OmpaP_link primers. Alpha fragment of β-lactamase amplified from pUC19 plasmid with AlphaL_link and AlphaP_XB primers. Resulting fragments joined by PCR using OmpaL_N and AlphaP_XB primers. Resulting fragment cutted with NdeI and BamHI and ligated into pACYC177_plac [notebook entries and another] Usage: Negative control in all of 'hunter and prey' tests


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
ACCACCTCCTCCAGATCCTCCGCCACCGCTTCCACCACTGCGGCCGCCAGAGCTCCCTCC
TCCAGAACCTCCGCCGTTGTCCGGACGAGTGCCGATGGTGTGTGCGTCACCGATGTTGTT
GGTCCACTGGTATTCCAGACGGGTAGCGATTTCAGGAGTGATCGCGTACTCAACACCGCC
AGCGAAGACCGGAGAAACGCCGGTGTCGTGGTTTTTACCATAAACGTTGGATTTAGTGTC
TGCACGCCATACCATGCCACCCAGACGAGTGTAGATGTCCAGGTCGTCAGTGATTGGGTA
ACCCAGTTTAGCGGTCAGTTGAACGCCCTGAGCTTTGTATGCACCGTTTTCAACGCTGCC
TTTGTACGGCATACGACCTAACCAGTCGTAACCCATTTCAAAGCCAACATACGGGTTAAC
CTGGTAACCACCAAAAGCACCAGCGCCCAGTTGGTTTTCATGGGTCGGGCCATTGTTGTT
GCCCAGTACCAGTTTAGTAGCTTTCA