pRF4 sequence - based on Mello et al.; orientation of insert confirmed by digest

Model Organism: Worms Methods & Reagents
1

AATTCTTATCATCTTCGGTTTTGATAAAATTGTGGTGTAGTCCATAATGAAATAAAATTAGAAACAATTGAAAATGAGAATAATTGGAAGGAAAGAGAACCGGTCATTTAGAATGAACACAATAGCTTTTTTAGTGATTGTTGCGTAAATTTTATGACATTACGTCATTGAATCCTGAAGAAGACAATGTTTAAATGTTCTCCAGATGGAATTAAATGTATAATTAAGCTCGAGATTTTTATCCGAGTTTTTATAAAAGTTGGTGAGACCCAGGATTAATCAAAAACAAGTATTGACAGTTCCGTCATTCATAACTCCACCGACTGCTCAAGTTGGCAAAAGTCAACTTTTAAAGTGGGTTATAAAGAGACTTCGATTTTTCAGATTTGCAAATCATTTGAAAGCATTTGTTAAACTAGATATAAATACATAGTTTTCCTCAATCAGGAAAATCAAACCACTGATCATCTGATCAGCTGTCTTCTAACAGGATGCACTATTAATGAATTAAACGAACTCGGTCTATTCACTCGGATAATTGCAACAATGTTAGTTCTATTTGATTTATTTCGGCGCGATCCGTAGCGACACTTTCTTCCAGCAACCATGTGATGCTGATTTCTATCATCCGAAGAGAGAAGGAAACATGGAGAGGAGCGGTTCAAGAACAGGTGAATCTGAACCTCCTTTTCCGTGTGTGTGGAAATATACACAAACAAGTGACTGATGAAGGCGGGAAAAAGAAAAACGATGGATTGAGTTATCTGGGTGAAGAATCTTAGACAAAGAGTTTGCAGAAAAAGCTTGTTCTTGGAAAAATAAAAAACTAGACAATTTTTAAAAAGGAAAACTACTTATTTATCACTTCAAAACCAGAACCCTTTTGACAATCTTTAACTGTTTGAACTTCGTATTAGATCTCAGCAGCAAAATAATAGTTGAAAAAATTTTATAAGAAAAGTTTTCGTGTTCAGTTGAAAACATCCAAAACAGAAAGTATTTTTATTTACTGATAATTCCTTAATACACATCTATTACCGTATTTGGGGCTTCTAAAAATATATTCTAGTCGTTTCAAAGTTCAAGCACATTACGTTAATTCATAGATCAAATCCACTAAACCTCAACCATCACTTTTTCTAATTTCTTCCACTTCAAAAAAAAACTAATCATCGTGGCTTCTTCCCGTTCCATGTCATATGGTCAACATTAATCAGTCGATGGAATTGGCCTTTTTACCGAAAATACGAAGATTCCACGCATCATTGCAACATTTTCGTCATAACCTCGCTAGCCCAAATCGGTTGTAGGATCCCCGAATCGCATTGTGGTTGAGCGTATGTCCTTCAAAAAGTCCAACAAAAACTCGAATACCGTTTGCCATCGGATGGTGTTGTTGAAGAGTTTTAACTATAAGCTACGCCCATCCGGACGTACGCGAGAAGCCCTTTCTGTTGAATGGGTGGGCGGCGGAAGAAAAAACTTCTTAACACGGCTTTTCGGTTTTTCTACCCTATATATTGGAGGATAGGTGAAGGATATTTTGGAGAATATACGATTAGCGGCAATCACAATGACATCCTTATGTATATGCATTCTTTTTATTGTTTCCTGATATTTATTCTCTCTTTATCATTTCACTTTTTCTTGGCACTCGTCCAAAAATCTCAATTAAAAATTTCTGGATATATCTTTAGATCTAACTGAAAATTTCCAGATGACCCTAACTACGGCGACGTCCGGCGCCATTGTATTTTCTGGAGCCACTTTGTTGGTTTCTCTTTTTGCCGCTGCTTCGCTTTACAGTCAAGTTTCCAATATCTGGAATGAGCTGGATGCCGAAATCGCCAACTTCAGAGTGAGTTTCCTATATAGCATTTCTAATTCACTGATAATTTCAATTATTTAGAGTCTCACTGAAGACATGTGGGTTGATATGGTTAAACTTGGAGCAGGAACCGCTTCCAACCGTGTGAGACGTCAACAATATGGAGGATATGGAGCCACTGGTGTTCAGCCACCAGCACCAACTCCAAACCCATATGGAGGATATGGAGCAAGCCAGCCAGCTCCACCAGAGAAATTCCCAGATGGTATACCAAATGGAGGAAATCAACCAAAGTTCCCAGGAGGTGGATTCCCAGATGGTCCATTCCCGAATGGAGGAGGACCACGTGGAGGAAATCAATGTCAATGCACTGTTGAGAACTCATGCCCACCAGGACCAGCTGGACCAGAAGGAGAGGAAGGACCAGATGGGCACGATGGACAAGACGGAGTCCCAGGATTTGACGGAAAAGATGCCGAAGATGTTCAAAACACCCCACCAACAGGATGCTTCACCTGTCCACAAGGACCACTTGGACCACAAGGACCAAATGGAGCTCCAGGACTCAGAGGAATGCGCGGAGCTCGTGGACAACCAGGACGTCCAGGAAGAGACGGAAACCCAGGAATGCCAGGAGACTGTGGACCACCAGGGGCCCCAGGATCCGATGGAAAGCCAGGATCTCCAGGAGGAAAGGGAGACGATGGAGAGAGACCATTGGGCCGCCCAGGACCAAGAGGACCACCAGGAGAGGCTGGGCCAGAAGGACCACAGGGACCAACTGGAAGAGATGCTTATCCAGGACAGTCTGGACCACAAGGAGAGCCAGGCCTTCAAGGATATGGAGGAGCTGCTGGAGAGGACGGGTAGGATGAAAAAAATATTAAACGACTTCTTGTAAATATAAAAATTTCAGACCAGAAGGACCACCAGGAGCCCCAGGACTTCCAGGAAAAGATGCCGAATACTGCAAATGCCCAGGAAGAGAAGGAGATGCTGGACGAAGTGCCAGACGTCATCGCAAATTCCAATTGTAGACAAATTCATGACATTTTTCCAAAAAATAAAACATACTTCTCAAAATTTTTTGTTTTGTATTTGGTTTACCATGGATGTTAAGAACTTTCTTGGGGAAAATAATTAAAATACAAAACTGTATAAATTATGTCACAAGCTTGGAAGTGCATTTGCAAAAAAACACTGGTGTAGAAGAACAAGAGGTTAGTATGACTTGAACAGAATAACTAATAGAAATTGGAGAGCATCCTTTTAGAAGGAAAAATGCACGACCTAGAACAGCTATTAATCTGTATTTGGCACTCGATTTATATTGCTAGAAATGGAACCTATTAGTATAAGGAAAAATAAGAGAAAAGGTGAGAAGGTCGGAAAAAACACTTCATGAATATATTTAGACGAGGAAAAAACGAAGGAAATAATGGGAGAGTGACGATAGAAAGAAGGAAAACGTTGAAAACTGAGTCGAAAAGAAAGAAAAGAACGAAGGTGAACAAAAACGACAAAGTTTTGAAAATCATTTCAATTTTCAGATTTCAGTTCAAGACCGAGTAGTTGGAAAACATGTGACCTCAAAAAAAAAAACAAAAAATTTCACTTAAAGGGGAAAGAAGTGAAGTAGAACTTCTGCATAATCATCAAAAGATTCCCGAATAGCTTGGTTCCAAAAGTTCAAACACAAATTACTTTTGGAAACTTTCATTTGTTTTGAACTGTAGCGGCATGTCGATTTTTTTCAAAATACAAATTACAATAGAAAATCAGGAAAACAATTGGAAGTGATTTTGGAAAAAAAAACGTTGAGGAGATTTGAGCACGAGACAAACAATTTCTGTAAATTTCAGTTCAAAAGCTTGAAAACGGTTCTTTCTTGCAAACAAAACCTAGTACCTCTTCACTTTAACATACACTTCACGAACAAGTTTTCTTTTGAAAATAATTTCTGAAAATCACGTTAACGAATCTCAAAATTCAGGAAATGCTCAAAAATGTCTATTTGCGTACTACGAAATTCCCGAAAAAACATCTGTGACTTTTTTCTCTCTTTTTGTTCGAAAAAATAACTTATGTCGCCGTGGTGGTGCCCTCGGGAATCGGAAAGCGTCAATGTTGTGTTTGTTGTTGTTATCCCAAACGAGAAGATGAATAAGAGGAGGAAAGATGAACGGAAAGCGAGAAAAAAAGAAGATGGAACACGTACATGAGCTTCTCCTGCTTTGAATACGAATTcgccctatagtgagtcgtattacaattcactggccgtcgttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaatcgccttgcagcacatccccctttcgccagctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaacagttgcgcagcctgaatggcgaatggcgcctgatgcggtattttctccttacgcatctgtgcggtatttcacaccgcatatggtgcactctcagtacaatctgctctgatgccgcatagttaagccagccccgacacccgccaacacccgctgacgcgccctgacgggcttgtctgctcccggcatccgcttacagacaagctgtgaccgtctccgggagctgcatgtgtcagaggttttcaccgtcatcaccgaaacgcgcgagacgaaagggcctcgtgatacgcctatttttataggttaatgtcatgataataatggtttcttagacgtcaggtggcacttttcggggaaatgtgcgcggaacccctatttgtttatttttctaaatacattcaaatatgtatccgctcatgagacaataaccctgataaatgcttcaataatattgaaaaaggaagagtatgagtattcaacatttccgtgtcgcccttattcccttttttgcggcattttgccttcctgtttttgctcacccagaaacgctggtgaaagtaaaagatgctgaagatcagttgggtgcacgagtgggttacatcgaactggatctcaacagcggtaagatccttgagagttttcgccccgaagaacgttttccaatgatgagcacttttaaagttctgctatgtggcgcggtattatcccgtattgacgccgggcaagagcaactcggtcgccgcatacactattctcagaatgacttggttgagtactcaccagtcacagaaaagcatcttacggatggcatgacagtaagagaattatgcagtgctgccataaccatgagtgataacactgcggccaacttacttctgacaacgatcggaggaccgaaggagctaaccgcttttttgcacaacatgggggatcatgtaactcgccttgatcgttgggaaccggagctgaatgaagccataccaaacgacgagcgtgacaccacgatgcctgtagcaatggcaacaacgttgcgcaaactattaactggcgaactacttactctagcttcccggcaacaattaatagactggatggaggcggataaagttgcaggaccacttctgcgctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtgggtctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatactttagattgatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacacaggaaacagctatgaccatgattacgccaagctcggaattaaccctcactaaagggaacaaaagcttgcatgcctgcaggtcgactctagaggatccccgggtaccgagctcg

2

Your sequence is almost right, but you have the wild-type rol-6 allele and not the su1006 substitution present in pRF4:

…gagcaggaaccgcttccaaccgtgtgaga C gtcaacaatatggaggatatggagccact… – Wild type
…gagcaggaaccgcttccaaccgtgtgaga T gtcaacaatatggaggatatggagccact… – su1006

The base in question is at position 1979 in the sequence you posted.

MM

3

thanks MM!