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

From 2008.igem.org

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Latest revision as of 16:55, 28 October 2008

pACYC177_lac + ompA-omega

Description

Author: Michał K.
Based on: pACYC177_plac
Resistance: kanamycin
Features: Fusion of OmpA and omega 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 OmegaL_link and OmegaP_EPB primers. Resulting fragments joined by PCR using OmpaL_N and OmegaP_EPB 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 GGGCAAATGGTGGTCACCATCCTGTCGGCTGTGGCACAGGCTGAACGCCGGAGGATCCAA CTGCAGAAGAATTCTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTA TTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGC TTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGAT TTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTA TCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGACCTCCACCT CCAGAACCACCTCCTCCAGATCCTCCGCCACCGCTTCCACCACTGCGGCCGCCAGAGCTC CCTCCTCCAGAACCTCCGCCGTTGTCCGGACGAGTGCCGATGGTGTGTGCGTCACCGATG TTGTTGGTCCACTGGTATTCCAGACGGGTAGCGATTTCAGGAGTGATCGCGTACTCAACA CCGCCAGCGAAGACCGGAGAAACGCCGGTGTCGTGGTTTTTACCATAAACGTTGGATTTA GTGTCTGCACGCCATACCATGCCACCCAGACGAGTGTAGATGTCCAGGTCGTCAGTGATT GGGTAACCCAGTTTAGCGGTCAGTTGAACGCCCTGAGCTTTGTATGCACCGTTTTCAACG CTGCCTTTGTACGGCATACGACCTAACCAGTCGTAACCCATTTCAAAGCCAACATACGGG TTAACCTGGTAACCACCAAAAGCACCAGCGCCCAGTTGGTTTTCATGGGTCGGGCCATTG TTGTTGCCCAGTACCAGTTTAGTAGCTTTCA

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+{{WarHeadVect|headimg=2008.igem.org/wiki/images/8/8d/Logo_project.jpg}}
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 <h2>pACYC177_lac + ompA-omega</h2>
+<p class=vector>Description</p><br>
+<b>Author</b>: Michał K.<br>
+<b>Based on</b>: <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177>pACYC177_plac</a><br>
+<b>Resistance</b>: kanamycin<br>
+<b>Features</b>: Fusion of OmpA and omega fragment of &beta;-lactamase under control of IPTG inducible lactose promoter. Low (10) copy plasmid<br>
+<b>Preparation</b>: OmpA fragment amplified from <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pB30D>pB30D</a> plasmid with
+<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>
+ 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#OmegaL_link">OmegaL_link</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmegaP_EPB">OmegaP_EPB</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#OmegaP_EPB">OmegaP_EPB</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>
+<b>Usage:</b> Negative control in all of 'hunter and prey' tests <br>
+<br><br>
+<p class=vector>Vector map</p><br><br>
 <img src="https://static.igem.org/mediawiki/2008/7/7a/PACYClac%2BompA-omega.png" width=600>
 <br><br>
-vector sequence:
+<p class=vector>Vector sequence</p><br><br>
 <pre>