PIJ12742

pIJ12742

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