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

From 2008.igem.org

(Difference between revisions)

Latest revision as of 16:54, 28 October 2008

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

@@ Line 12: / Line 12: @@
 <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#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>]</b<br>
+  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>
 <b>Usage:</b> Negative control in all of 'hunter and prey' tests <br>
 <br><br>