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Latest revision as of 16:38, 8 August 2019
pIJ12742
Background
Construction
Uses
Selecting for gene deletions in actinomycetes
Characteristics
Contains the codon optimised I-SceI gene under the control of the strong constitutive ermE* promoter
Derivative of pGM1190, an intermediate copy number, conjugative plasmid containing the temperature-sensitive replication origin of pSG5
Map and Sequence
ggccgtcgacgacgacgcgtccgcctgcctgcttttggccggcgggccgggccgcggttgtccgccgggtcggaccggtcccttggtcgaccgatgcctcggccacgacgatgctgggcccgaagcggtacgcgaacccgtgccgggctgcggcggatcgcggcagcaggagctccccgcccgccacggctggctggatggtcgcctcacgcgcccgcaggggtgacggcgtgggtgcggcggtcagccccgtgatcgggcgtcagacggccgctgagggaccgcgaccacccctcccttcccgcctgccgtctttcaccgcccggccggtccgcagacgccgtacggccaggccacggaggattacgggcagatggcgcccacccggcacccgggctggggcaccctggacagcacgaaggcccgccccactcaccggggcgggccttcgcgttgagctagagttcccgccagcctcgcagagcaggattcccgttgagcaccgccaggtgcgaataagggacagtgaagaaggaacacccgctcgcgggtgggcctacttcacctatcctgcccggctgacgccgttggatacaccaaggaaagtctacacgaaccctttggcaaaatcctgtatatcgtgcgaaaaaggatggatataccgaaaaaatcgctataatgaccccgaagcagggttatgcagcggaaaatgcagctcacggtaactgatgccgtatttgcagtaccagcgtacggcccacagaatgatgtcacgctgaaaatgccggcctttgaatgggttcatgtgcagctccatcagcaaaaggggatgataagtttatcaccaccgactatttgcaacagtgccgttgatcgtgctatgatcgactgatgtcatcagcggtggagtgcaatgtcgtgcaatacgaatggcgaaaagccgagctcatcggtcagcttctcaaccttggggttacccccggcggtgtgctgctggtccacagctccttccgtagcgtccggcccctcgaagatgggccacttggactgatcgaggccctgcgtgctgcgctgggtccgggagggacgctcgtcatgccctcgtggtcaggtctggacgacgagccgttcgatcctgccacgtcgcccgttacaccggaccttggagttgtctctgacacattctggcgcctgccaaatgtaaagcgcagcgcccatccatttgcctttgcggcagcggggccacaggcagagcagatcatctctgatccattgcccctgccacctcactcgcctgcaagcccggtcgcccgtgtccatgaactcgatgggcaggtacttctcctcggcgtgggacacgatgccaacacgacgctgcatcttgccgagttgatggcaaaggttccctatggggtgccgagacactgcaccattcttcaggatggcaagttggtacgcgtcgattatctcgagaatgaccactgctgtgagcgctttgccttggcggacaggtggctcaaggagaagagccttcagaaggaaggtccagtcggtcatgcctttgctcggttgatccgctcccgcgacattgtggcgacagccctgggtcaactgggccgagatccgttgatcttcctgcatccgccagaggcgggatgcgaagaatgcgatgccgctcgccagtcgattggctgagctcataagttcctattccgaagttcctattcgaaatgaccgaccaagcgacgcccaacctgccatcacgagatttcgattccaccgccgccttctatgaaaggttgggcttcggaatcgttttccgggacgccggctggatgatcctccagcgcggggatctcatgctggagttcttcgcccaccccaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacacaggaaacagctatgaccatgattacgaattgtacgtaggatccagcccgacccgagcacgcgccggcacgcctggtcgatgtcggaccggagttcgaggtacgcggcttgcaggtccaggaaggggacgtccatgcgagtgtccgttcgagtggcggcttgcgcccgatgctagtcgcggttgatcggcgatcgcaggtgcacgcggtcgatcttgacggctggcgagaggtgcggggaggatctgaccgacgcggtccacacgtggcaccgcgatgctgttgtgggcacaatcgtgccggttggtaggatcgtctagaacaggaggccccatatgcaccagaagaaccaggtcatgaacctgggcccgaactccaagctgctgaaggagtacaagtcccagctgatcgagctgaacatcgagcagttcgaggccggcatcggcctgatcctgggcgacgcctacatccgctcccgcgacgagggcaagacctactgcatgcagttcgagtggaagaacaaggcctacatggaccacgtctgcctgctgtacgaccagtgggtcctgtccccgccgcacaagaagcagcgcgtcaaccacctgggcaacctggtcatcacctggggcgcccagaccttcaagcaccaggccttcaacaagctggccaacctgttcatcgtcaacaacaagaagaccatcccgaacaacctggtcgagaactacctgaccccgatgtccctggcctactggttcatggacgacggcggcaagtgggactacaacaagaactccaccaacaagtccatcgtcctgaacacccagtccttcaccttcgaggaggtcgagtacctggtcaagggcctgcgcaacaagttccagctgaactgctacgtcaagatcaacaagaacaagccgatcatctacatcgactccatgtcctacctgatcttctacaacctgatcaagccgtacctgatcccgcagatgatgtacaagctgccgaacaccatctcctccgagaccttcctgaagtgataaaccgatacaattaaaggctccttttggagcctttttttttggagattttcaacgtgaaaaaattattattcgcaattccaacaaaaggctggagctccaccgcggccgcgcgcgatatcgaattccccagatctaaagttttgtcgtctttccagacgttagtaaatgaattttctgtatgaggttttgctaaacaactttcaacagtttcagcggagtgagaatagaaaggaacaactaaaggaattgcgaataataattttttcacgttgaaaatctccaaaaaaaaaggctccaaaaggagcctttaattgtatcggtttatcagcttgctttcgaggtgaatttcttaaacagcttgataccgatagttgcgccgacaatgacaacaaccatcgcccacgcataaccgatatattcggtcgctgaggcttgcagggagtcaaaggccgcttttgcgggatctcgtcgaaggcactagaaggcctatatgcggggatcgaccgcgcgggtcccggacggggaagagcggggagctttgccagagagcgacgacttccccttgcgttggtgattgccggtcagggcagccatccgccatcgtcgcgtagggtgtcacaccccaggaatcgcgtcactgaacacagcagccggtaggacgaccatgactgagttggacaccatcgcaaatccgtccgatcccgcggtgcagcggatcatcgatgtcaccaagccgtcgcgatccaacataaagacaacgttgatcgaggacgtcgagcccctcatgcacagcatcgcggccggggtggagttcatcgaggtctacggcagcgacagcagtccttttccatctgagttgctggatctgtgcgggcggcagaacataccggtccgcctcatcgactcctcgatcgtcaaccagttgttcaagggggagcggaaggccaagacattcggcatcgcccgcgtccctcgcccggccaggttcggcgatatcgcgagccggcgtggggacgtcgtcgttctcgacggggtgaagatcgtcgggaacatcggcgcgatagtacgcacgtcgctcgcgctcggagcgtcggggatcatcctggtcgacagtgacatcaccagcatcgcggaccggcgtctccaaagggccagccgaggttacgtcttctcccttcccgtcgttctctccggtcgcgaggaggccatcgccttcattcgggacagcggtatgcagctgatgacgctcaaggcggatggcgacatttccgtgaaggaactcggggacaatccggatcggctggccttgctgttcggcagcgaaaagggtgggccttccgacctgttcgaggaggcgtcttccgcctcggtttccatccccatgatgagccagaccgagtctctcaacgtttccgtttccctcggaatcgcgctgcacgagaggatcgacaggaatctcgcggccaaccgataagcgcctctgttcctcggacgctcggttcctcgacctcgattcgtcagtgatgatcaacaggacccagaaatggcacgagcccgggattggcctcccgggctcgccgcccccgaaccggcgcagcatgccgagtgaggacgtttgctctgtgacgagcgtacgagagaacagcgcgtcgacaccaccgcgccccgccggaccgaagtgccactgcggcaccccgatcgtccacatccccgggaagcgccccaaaatctactgctcggacgcctgccgaaagcgcgcaaagcgtctcattgcggaccagtcacgcagcgtagcaagcacgactacagcgggtaatccggacgcgggctttagtagtaagggaaaatctccgtccactgcaggtcagagcgctgtaactggaacgggttctaccccaaacgagggtgccgcgacagccgggcgggaggacccgcgcgacgagcggttcgcccggcgggaccggcaccagacggtgtcgctgaacgaggcgttcaagggctgcgggacccggctcactgccgggacggccgagctgatgtggaagccgggcgaggcgacgtggggcaacacgtgccgctgcaacaacgtccacacctgcccgtggtgcatgagccgcatcctggccgtacggggctcgaacgtgcagctcgccgccgacggtctcgcggacgccgggtacgggctgcaccttggcacgaacacgttgcgccacttcgagcgcatggcgttcggcaccgtccgcaagggaatgcggcacggcctggtcgcggtcttgcacgacggctggaagggcgcgtacggctcgtccggccgccgctggcgcacgatgcgcgacgacttcgggatcatcggctacgagcgggccttcgaggacaccttcggctggggctccggctggcacctgcactggcacacgctctgggtcacccgcgaggtgctcggcccggacgcccaggccgcgttccgcgacgcactcgccggagcctgggccgccggggtcgaatccgcgggcggctacaccgtcagcgagacgtgcgaccggcccggttgctcctgtgagggcaagggccacggcaccgacgtgcgcccactcaacggcgcggacgccgctgacggcgatgcgggcaagcaggcccgctacctctacaaggacggcgacaagaccaagggcggcgtcgccaagatcggcctggagctggccggacagaacttcaaggccggtcgcggcgacgaccgcatgggaccgctcgacctgggcgacgcggcggccgccgagctgcagcggcttcgtcggcccggcccgttcgtcgagaagtaccgcgagcgcgagttcggcgtcttccaggtccgcaagcactaccggtcgcagaacctgaaccggctgatcaaggaactcggcatccagcaggacgtccgcaccgaggaggagatcaccgacgacaccgagggcctggtcgccatcgcggtcatcccggcctacatctggtaccgatacatcgcccgcgtcgccggacgtcgcctcgacctcatcaaggtcgccgagacgtacggcctaccgggcgtccggcggctcgtcgagtcctggggcctggtgtggggcaaggacgtcctggacccgcccgccccggaagcgccggccgcgccgggcgaccttgacgctgaccagatgaggttcgaggtcatgtccgaggaggaagctgctttccgcgaggcgcgccggaaggccaacgaggcccgcacggaggagctggccgcttcgctcgaccgagtgcgtcagccgaagaaggaggcgatccgcccgacgatcagccttcggaagcgcctgaagccgaagccggtcaccgtcgacgtcaagactcccccgcccggcgccgcgtcgcccgtgtgccgccgctgcaagggcaagctcgccccggtactccagccctggggccggctccccggagactgcctccgcgtcgacacagcggtcgcgtagggccgctgagggaccgccatgagaccgccaccccggacatccgggtggcggtctcatggcggtcccggcgcgaaagccgagggtctgcctgccgtgaggtggccggcggacctgggttactgctggaatcccgcgcccagtccgagcgtccgccggcgcttttcacctggctccgggtgctcgtcgatgagcacca
References
Fernández-Martínez LT, Bibb MJ. Use of the meganuclease I-SceI of Saccharomyces cerevisiae to select for gene deletions in actinomycetes. Sci Rep. 2014;4:7100. Published 2014 Nov 18. doi:10.1038/srep07100