Difference between revisions of "PMAD"

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<pubmed> 15528558 </pubmed>
 
<pubmed> 15528558 </pubmed>
  
'''The sequence'''
+
'''Sequencing oligos'''
*  
+
*
GAATTAATTCTCATGTTTGACAGCTTATCATCGATTCACAAAAAATAGGC
 
ACACGAAAAACAAGTTAAGGGATGCAGTTTATGCATCCCTTAACTTACTT
 
ATTAAATAATTTATAGCTATTGAAAAGAGATAAGAATTGTTCAAAGCTAA
 
TATTGTTTAAATCGTCAATTCCTGCATGTTTTAAGGAATTGTTAAATTGA
 
TTTTTTGTAAATATTTTCTTGTATTCTTTGTTAACCCATTTCATAACGAA
 
ATAATTATACTTTTGTTTATCTTTGTGTGATATTCTTGATTTTTTTCTAC
 
TTAATCTGATAAGTGAGCTATTCACTTTAGGTTTAGGATGAAAATATTCT
 
CTTGGAACCATACTTAATATAGAAATATCAACTTCTGCCATTAAAAGTAA
 
TGCCAATGAGCGTTTTGTATTTAATAATCTTTTAGCAAACCCGTATTCCA
 
CGATTAAATAAATCTCATTAGCTATACTATCAAAAACAATTTTGCGTATT
 
ATATCCGTACTTATGTTATAAGGTATATTACCATATATTTTATAGGATTG
 
GTTTTTAGGAAATTTAAACTGCAATATATCCTTGTTTAAAACTTGGAAAT
 
TATCGTGATCAACAAGTTTATTTTCTGTAGTTTTGCATAATTTATGGTCT
 
ATTTCAATGGCAGTTACGAAATTACACCTCTTTACTAATTCAAGGGTAAA
 
ATGGCCTTTTCCTGAGCCGATTTCAAAGATATTATCATGTTCATTTAATC
 
TTATATTTGTCATTATTTTATCTATATTATGTTTTGAAGTAATAAAGTTT
 
TGACTGTGTTTTATATTTTTCTCGTTCATTATAACCCTCTTTAATTTGGT
 
TATATGAATTTTGCTTATTAACGATTCATTATAACCACTTATTTTTTGTT
 
TGGTTGATAATGAACTGTGCTGATTACAAAAATACTAAAAATGCCCATAT
 
TTTTTCCTCCTTATAAAATTAGTATAATTATAGCACGAGCTCTGATAAAT
 
ATGAACATGATGAGTGATCGTTAAATTTATACTGCAATCGGATGCGATTA
 
TTGAATAAAAGATATGAGAGATTTATCTAATTTCTTTTTTCTTGTAAAAA
 
AAGAAAGTTCTTAAAGGTTTTATAGTTTTGGTCGTAGAGCACACGGTTTA
 
ACGACTTAATTACGAAGTAAATAAGTCTAGTGTGTTAGACTTTATGAAAT
 
CTATATACGTTTATATATATTTATTATCCGGAGGTGTAGCATGTCTCATT
 
CAATTTTGAGGGTTGCCAGAGTTAAAGGATCAAGTAATACAAACGGGATA
 
CAAAGACATAATCAAAGAGAGAATAAAAACTATAATAATAAAGACATAAA
 
TCATGAGGAAACATATAAAAATTATGATTTGATTAACGCACAAAATATAA
 
AGTATAAAGATAAAATTGATGAAACGATTGATGAGAATTATTCAGGGAAA
 
CGTAAAATTCGGTCAGATGCAATTCGACATGTGGACGGACTGGTTACAAG
 
TGATAAAGATTTCTTTGATGATTTAAGCGGAGAAGAAATAGAACGATTTT
 
TTAAAGATAGCTTGGAGTTTCTAGAAAATGAATACGGTAAGGAAAATATG
 
CTGTATGCGACTGTCCATCTGGATGAAAGAGTCCCACATATGCACTTTGG
 
TTTTGTCCCTTTAACAGAGGACGGGAGATTGTCTGCAAAAGAACAGTTAG
 
GCAACAAGAAAGACTTTACTCAATTACAAGATAGATTTAATGAGTATGTG
 
AATGAGAAAGGTTATGAACTTGAAAGAGGCACGTCCAAAGAGGTTACAGA
 
ACGAGAACATAAAGCGATGGATCAGTACAAGAAAGATACTGTATTTCATA
 
AACAGGAACTGCAAGAAGTTAAGGATGAGTTACAGAAGGCAAATAAGCAG
 
TTACAGAGTGGAATAGAGCATATGAGGTCTACGAAACCCTTTGATTATGA
 
AAATGAGCGTACAGGTTTGTTCTCTGGACGTGAAGAGACTGGTAGAAAGA
 
TATTAACTGCTGATGAATTTGAACGCCTGCAAGAAACAATCTCTTCTGCA
 
GAACGGATTGTTGATGATTACGAAAATATTAAGAGCACAGACTATTACAC
 
AGAAAATCAAGAATTAAAAAAACGTAGAGAGAGTTTGAAAGAAGTAGTGA
 
ATACATGGAAAGAGGGGTATCACGAAAAAAGTAAAGAGGTTAATAAATTA
 
AAGCGAGAGAATGATAGTTTGAATGAGCAGTTGAATGTATCAGAGAAATT
 
TCAAGCTAGTACAGTGACTTTATATCGTGCTGCGAGGGCGAATTTCCCTG
 
GGTTTGAGAAAGGGTTTAATAGGCTTAAAGAGAAATTCTTTAATGATTCC
 
AAATTTGAGCGTGTGGGACAGTTTATGGATGTTGTACAGGATAATGTCCA
 
GAAGGTCGATAGAAAGCGTGAGAAACAGCGTACAGACGATTTAGAGATGT
 
AGAGGTACTTTTATGCCGAGAAAACTTTTTGCGTGTGACAGTCCTTAAAA
 
TATACTTAGAGCGTAAGCGAAAGTAGTAGCGACAGCTATTAACTTTCGGT
 
TGCAAAGCTCTAGGATTTTTAATGGACGCAGCGCATCACACGCAAAAAGG
 
AAATTGGAATAAATGCGAAATTTGAGATGTTAATTAAAGACCTTTTTGAG
 
GTCTTTTTTTCTTAGATTTTTGGGGTTATTTAGGGGAGAAAACATAGGGG
 
GGTACTACGACCTCCCCCCTAGGTGTCCATTGTCCATTGTCCAAACAAAT
 
AAATAAATATTGGGTTTTTAATGTTAAAAGGTTGTTTTTTATGTTAAAGT
 
GAAAAAAACAGATGTTGGGAGGTACAGTGATAGTTGTAGATAGAAAAGAA
 
GAGAAAAAAGTTGCTGTTACTTTAAGACTTACAACAGAAGAAAATGAGAT
 
ATTAAATAGAATCAAAGAAAAATATAATATTAGCAAATCAGATGCAACCG
 
GTATTCTAATAAAAAAATATGCAAAGGAGGAATACGGTGCATTTTAAACA
 
AAAAAAGATAGACAGCACTGGCATGCTGCCTATCTATGACTAAATTTTGT
 
TAAGTGTATTAGCACCGTTATTATATCATGAGCGAAAATGTAATAAAAGA
 
AACTGAAAACAAGAAAAATTCAAGAGGACGTAATTGGACATTTGTTTTAT
 
ATCCAGAATCAGCAAAAGCCGAGTGGTTAGAGTATTTAAAAGAGTTACAC
 
ATTCAATTTGTAGTGTCTCCATTACATGATAGGGATACTGATACAGAAGG
 
TAGGATGAAAAAAGAGCATTATCATATTCTAGTGATGTATGAGGGTAATA
 
AATCTTATGAACAGATAAAAATAATTAACAGAAGAATTGAATGCGACTAT
 
TCCGCAGATTGCAGGAAGTGTGAAAGGTCTTGTGAGATATATGCTTCACA
 
TGGACGATCCTAATAAATTTAAATATCAAAAAGAAGATATGATAGTTTAT
 
GGCGGTGTAGATGTTGATGAATTATTAAAGAAAACAACAACAGATAGATA
 
TAAATTAATTAAAGAAATGATTGAGTTTATTGATGAACAAGGAATCGTAG
 
AATTTAAGAGTTTAATGGATTATGCAATGAAGTTTAAATTTGATGATTGG
 
TTCCCGCTTTTATGTGATAACTCGGCGTATGTTATTCAAGAATATATAAA
 
ATCAAATCGGTATAAATCTGACCGATAGATTTTGAATTTAGGTGTCACAA
 
GACACTCTTTTTTCGCACCAGCGAAAACTGGTTTAAGCCGACTGCGCAAA
 
AGACATAATCGATAAGCTTTAATGCGGTAGTTTATCACAGTTAAATTGCT
 
AACGCAGTCAGGCACCGTGTATGAAATCTAACAATGCGCTCATCGTCATC
 
CTCGGCACCGTCACCCTGGATGCTGTAGGCATAGGCTTGGTTATGCCGGT
 
ACTGCCGGGCCTCTTGCGGGATCGAGGCCTTTAGCATATTATGTTGCCAA
 
CTGTCGGAACGAGACTTCTCTAAACCTTCCCGGCTTCATCATGCTCTCTT
 
AACACTGCTACATCAACCCCTTGAATCCTCAATTCACCTCCTCGAAAACA
 
TTTCCCATCAATCATATTCTGGTATATCTTATCTTCTGGCAGTGACAGCG
 
TCACTTCGTAATCATTATGATTGATGATAATCAAATACTTCCATTCATCA
 
GTCTCTCTTTGTTGCACCTCTACATTTTCAGCTACTTCAAGAATGGGATT
 
AATATGATGTTTAGCGAAAACCTGTTCTAAAAGCCTGCCTAAATAATTAC
 
TATCTGGATAAGTACCGACGTAAATCCCCTCTCCTTTACCGTAGCAGTTA
 
CGTGTAACCGCCGGAAGTCCTGCATACCAATCCCCCTTAAATGTCGCTAG
 
AGGTTCTGCCCCTTCTAATCGGATTATGTCCGCCCACGTCGTACAATCAT
 
ATTCCCCATCGTTACTATATATTTTGTTTACCTTTGTTTCTGGGTATGGT
 
ACAAATTCCTCAACAAAAATCCCCAAAATATCTCGCAGAGGACCAGGATA
 
TCCGCCTAGATGTACACGGTCATTTTCATCTACAATGCCACTGAAGAAAC
 
TGACAATCAAAGTGCCACCGTTAGCAACAAATTGCCGTAAGTTTTCATCT
 
TCTCCCTCTTTAACCATATATAACATTGGAGCAATAACTACTTTGTATTT
 
TGTTAGATCATCAGATGGCCTTACAAAATCGACAGCAATATTACGTTTAT
 
ATAATTCCCTATAATAAGCTTCAACTATAGGAATATATCTTAGTTTATTA
 
TGTGGTTTGGAACTTAGTTCGACAGCCCACCAGTTTTCCCAATCAAAAAT
 
GATCGCGACCTCTGCCTTGATTCTAGATCCGACCAAACAATCTAACTTTT
 
TCAGCTCTTGCCCTAACTGTGTAACTTCCCTATAAATTCTATTATTCTCG
 
TTCAAAAAGTGGGGCACCATTGCACCGTGGAATTTTTCAGCTCCTGCTCT
 
ACTTTGACGCCACTGGAAAAACATAATACCATCTGCACCACGGGCAATAG
 
TTGCATAACTCCATAGACGCATTACACCTGGCGGTTTTGGAACATTAATA
 
TCGCGCCAGTTAACATGTGAGGTTACCTGCTCCATCAAAATAAACGGTTG
 
ACCTTTTCTTAAACTACGCATAAGGTCATTCATCATGGCGTGCTGAATTG
 
GCAAGCCCTCTCTGGGATCAGGATATGAGTCCCATGTCACAATATCTACA
 
TGCTGAGCCCATTGAAAATAGTTTAACGGTTTGAATGAACCCATGAAATT
 
AGTTGATACTGGAATATCTGGTGTTACCTCACGTAAAATTTCCTTTTCTG
 
TTAAAAACAACTTGAGAATTGAGTCATTCATAAAACGGTAGTAATCAAGT
 
TCTTGGGATGGATTAATAAAAGTTGGTGCCTTTCTAGGGGGATTAATTTC
 
ATCCCAATGATTGTATCGCTGTCCCCAAAAGTTTGTACCCCAACGTTCAT
 
TTAATTCATCGATTGTTTTATATCTTTCCTTTAGCCACTTTCTAAACGCG
 
ACAGCACAATTCTCACAAAAACACTTGGAAACGTGACATGCATACTCATT
 
ATTAACATGCCACATTTTGAGTGCCGGATGATTTTTATACCGTTCTGCTA
 
TAGCCCTCACAAGTCTCTTTATGTGCGTAATTAATTGAGGATGATTAGGA
 
CAATAATGTTGTCTACTGCCAAACGAGAGAATGACACCGCTTTCATCGAT
 
CGGCAAAGAATCTGGATACTTTTTTACAAACCAAGCTGGAGTAGTTGCAG
 
TCGCCGTCCCCAAGTTAATATAAACACCGTGGTCATATAGTATATCTATA
 
ACCTTGTCTAGCCATTCAAAGTCGAACACTCCATCAGACGGTTCGATCTT
 
GCTCCAACTGAAAATCCCTAAAGATACTAAATTCACCCCCGCTTTTTGCA
 
TCAACTTAGCATCTTCATACCAAATTTCCTCTGGCCATTGCTCTGGGTTA
 
TAATCTCCTCCGTAACAAATTGAGGATAACACATTCATAATTATTCCCCC
 
TAGCTAATTTTCGTTTAATTATAAATTAAGTTAAAATTTAGGTACCTACG
 
TAGGATCGATCCGATCCTTAATTATATTATAGTCCCAATATAATCATTTA
 
TCAACTCTTTTACACTTAAATTTCCTAATATTTGTAAATATTTTTTTGAA
 
TAAAAAAATCTAGCTAATGTTACGTTACACATTAACTAGACAGATCTATC
 
GATGCATGCCATGGTACCCGGGAGCTCGAATTCTAGAAGCTTCTGCAGAC
 
GCGTCGACGTCATATGGATCCGATATCGCCCGACGCGAGGCTGGATGGCC
 
TTCCCCATTATGATTCTTCTCGCTTCCGGCGGCATCGGGATGCCCGCGTT
 
GCAGGCCATGCTGTCCAGGCAGGTAGATGACGACCATCAGGGACAGCTTC
 
AAGGATCGCTCGCGGCTCTTACCAGCCTAACTTCGATCACTGGACCGCTG
 
ATCGTCACGGCGATTTATGCCGCCTCGGCGAGCACATGGAACGGGTTGGC
 
ATGGATTGTAGGCGCCGCCCTATACCTTGTCTGCCTCCCCGCGTTGCGTC
 
GCGGTGCATGGAGCCGGGCCACCTCGACCTGAATGGAAGCCGGCGGCACC
 
TCGCTAACGGATTCACCACTCCAAGAATTGGAGCCAATCAATTCTTGCGG
 
AGAACTGTGAATGCGCAAACCAACCCTTGGCAGAACATATCCATCGCGTC
 
CGCCATCTCCAGCAGCCGCACGCGGCGCATCTCGGGCAGCGTTGGGTCCT
 
GGCCACGGGTGCGCATGATCGTGCTCCTGTCGTTGAGGACCCGGCTAGGC
 
TGGCGGGGTTGCCTTACTGGTTAGCAGAATGAATCACCGATACGCGAGCG
 
AACGTGAAGCGACTGCTGCTGCAAAACGTCTGCGACCTGAGCAACAACAT
 
GAATGGTCTTCGGTTTCCGTGTTTCGTAAAGTCTGGAAACGCGGAAGTCA
 
GCGCCCTGCACCATTATGTTCCGGATCTGCATCGCAGGATGCTGCTGGCT
 
ACCCTGTGGAACACCTACATCTGTATTAACGAAGCGCTGGCATTGACCCT
 
GAGTGATTTTTCTCTGGTCCCGCCGCATCCATACCGCCAGTTGTTTACCC
 
TCACAACGTTCCAGTAACCGGGCATGTTCATCATCAGTAACCCGTATCGT
 
GAGCATCCTCTCTCGTTTCATCGGTATCATTACCCCCATGAACAGAAATT
 
CCCCCTTACACGGAGGCATCAAGTGACCAAACAGGAAAAAACCGCCCTTA
 
ACATGGCCCGCTTTATCAGAAGCCAGACATTAACGCTTCTGGAGAAACTC
 
AACGAGCTGGACGCGGATGAACAGGCAGACATCTGTGAATCGCTTCACGA
 
CCACGCTGATGAGCTTTACCGCAGCTGCCTCGCGCGTTTCGGTGATGACG
 
GTGAAAACCTCTGACACATGCAGCTCCCGGAGACGGTCACAGCTTGTCTG
 
TAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGGCGCGTCAGCGGGTGT
 
TGGCGGGTGTCGGGGCGCAGCCATGACCCAGTCACGTAGCGATAGCGGAG
 
TGTATACTGGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGC
 
ACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGC
 
ATCAGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGT
 
TCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTAT
 
CCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAG
 
CAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAG
 
GCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGT
 
GGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGC
 
TCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTC
 
CGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTA
 
GGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCAC
 
GAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCT
 
TGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTG
 
GTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTG
 
AAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTG
 
CGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGAT
 
CCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAG
 
CAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTC
 
TACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGG
 
TCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAA
 
TGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAG
 
TTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTC
 
GTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGG
 
GAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACG
 
CTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCG
 
AGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAAT
 
TGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAA
 
CGTTGTTGCCATTGCTGCAGGCATCGTGGTGTCACGCTCGTCGTTTGGTA
 
TGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCC
 
CCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGT
 
CAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGC
 
ATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGT
 
GAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTG
 
CTCTTGCCCGGCGTCAACACGGGATAATACCGCGCCACATAGCAGAACTT
 
TAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGG
 
ATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAA
 
CTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAA
 
CAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGT
 
TGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGG
 
TTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAAC
 
AAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAA
 
GAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAG
 
GCCCTTTCGTCTTCAA
 

Revision as of 09:23, 29 August 2012

pMAD: click to enlarge

General comments

This site is under construction

A plasmid for generating gene inactivation mutants in naturally nontransformable gram-positive bacteria. This vector allows, on X-Gal (5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside) plates, a quick colorimetric blue-white discrimination of bacteria which have lost the plasmid, facilitating clone identification during mutagenesis. The plasmid is can be used in Staphylococcus aureus, Listeria monocytogenes, and Bacillus spp. to efficiently construct mutants with or without an associated antibiotic resistance gene.

  • E.coli: multicopy plasmid, select for ampicillin
  • B. subtilis: temperature-sensitive ori, select for erythromycin/ lincomycin

Hints for the usage

  • for a markerless deletion of a gene amplify 1000 bp up- and downstream of the gene and clone them into the MCS of the plasmid

The reference

Maryvonne Arnaud, Arnaud Chastanet, Michel Débarbouillé
New vector for efficient allelic replacement in naturally nontransformable, low-GC-content, gram-positive bacteria.
Appl Environ Microbiol: 2004, 70(11);6887-91
[PubMed:15528558] [WorldCat.org] [DOI] (P p)


Sequencing oligos