View source for PIJ12742

=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