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

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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  
<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>
<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#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><br>
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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#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>]</b<br>
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<b>Usage:</b> <br>
+
<b>Usage:</b> Negative control in all of 'hunter and prey' tests <br>
<br><br>
<br><br>

Revision as of 16:47, 28 October 2008

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Vectors view

 

pACYC177_lac + ompA-omega

Description


Author: Michał K.
Date: 2 July 2008
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] 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