User:Mingjie Li/Notebook/Light switch/2008/11/13

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ML1-1
Sequence: ATGGTGAGCGGAGTGGGAGGCAGCGGAGGAGGAAGGGGCGGCGGAAGGGG AGGAGAGGAGGAGCCTAGCAGCAGCCACACCCCTAACAACAGGAGGGGAGGAGAGCAGGC TCAGAGCAGCGGAACCAAGAGCCTGAGGCCTAGGAGCAACACCGAGAGCATGAGCAAGGC CATCCAGCAGTACACCGTGGACGCCAGGCTGCACGCCGTGTTCGAGCAGAGCGGCGAGAG CGGCAAGAGCTTCGACTACAGCCAGAGCCTGAAGACCACCACCTACGGCAGCAGCGTGCC TGAGCAGCAGATCACAGCCTACCTGAGCAGAATCCAGAGGGGCGGCTACATCCAGCCTTT CGGCTGCATGATCGCCGTGGACGAGAGCAGCTTCAGGATCATCGGCTACAGCGAGAACGC CAGGGAGATGCTGGGCATCATGCCTCAGAGCGTGCCTACCCTGGAGAAGCCTGAGATCCT GGCCATGGGCACCGACGTGAGGAGCCTGTTCACCAGCAGCAGCAGCATCCTGCTGGAGAG GGCCTTCGTGGCCAGGGAGATCACACTGCTCAACCCTGTGTGGATTCACAGCAAGAACAC CGGCAAGCCTTTCTACGCCATCCTGCACAGGATCGACGTGGGCGTGGTCATCGACCTGGA GCCTGCTAGGACCGAGGACCCTGCCCTGAGCATCGCTGGAGCTGTGCAGAGCCAGAAGCT GGCCGTGAGGGCTATCAGCCAGCTGCAGGCCCTGCCTGGCGGCGACATCAAGCTGCTGTG CGACACCGTGGTGGAGAGCGTGAGGGACCTGACCGGCTACGACAGAGTGATGGTGTACAA GTTCCACGAGGACGAGCACGGCGAGGTGGTGGCCGAGAGCAAGAGGGACGACCTGGAGCC TTACATCGGCCTGCACTACCCTGCCACCGACATCCCTCAGGCCAGCAGGTTCCTGTTCAA GCAGAACAGGGTGAGGATGATCGTGGACTGCAACGCCACCCCTGTGCTCGTGGTGCAGGA CGACAGGCTGACCCAGAGCATGTGCCTGGTGGGCAGCACCCTGAGGGCCCCTCACGGCTG CCACAGCCAGTACATGGCCAACATGGGCAGCATCGCCAGCCTGGCCATGGCCGTGATCAT CAACGGCAACGAGGACGACGGCAGCAACGTGGCCAGCGGCAGGAGCAGCATGAGGCTGTG GGGCCTGGTGGTGTGCCACCACACCAGCAGCAGGTGCATCCCTTTCCCTCTGAGATACGC CTGCGAGTTCCTGATGCAGGCCTTCGGCCTGCAGCTGAACATGGAGCTGCAGCTGGCCCT GCAGATGAGCGAGAAGAGGGTGCTGAGGACCCAGACCCTGCTGTGCGACATGCTGCTGAG GGACAGCCCTGCCGGCATCGTGACCCAGAGCCCTAGCATCATGGACCTGGTGAAGTGCGA CGGCGCCGCCTTCCTGTACCACGGCAAGTACTACCCTCTGGGCGTGGCCCCTAGCGAGGT GCAGATCAAGGACGTGGTGGAGTGGCTGCTGGCCAACCACGCCGACAGCACCGGCCTGAG CACCGACAGCCTGGGCGACGCTGGATACCCTGGAGCTGCCGCCCTGGGCGACGCCGTGTG CGGCATGGCCGTGGCCTACATCACAAAGAGGGACTTCCTGTTCTGGTTCAGGAGCCACAC CGCCAAGGAGATCAAGTGGGGCGGCGCCAAGCACCACCCTGAGGACAAGGACGACGGCCA GAGGATGCACCCTAGGAGCAGCTTCCAGGCCTTCCTGGAGGTGGTGAAGAGCAGGAGCCA GCCTTGGGAGACCGCCGAGATGGACGCCATCCACAGCCTGCAGCTGATCCTGAGGGACAG CTTCAAGGAGAGCTGA
 * Optimize gene:
 * 1. Arabidopsis PHYB(NT): Length:1866;

Sequence: ATGCCTCTGTTCGAGCTGTTCAGGCTGACCAAGGCCAAGCTGGAGAGCGC CCAGGACAGGAACCCTAGCCCTCCTGTGGACGAGGTGGTGGAGCTGGTGTGGGAGAACGG CCAGATCAGCACCCAGAGCCAGAGCAGCAGGAGCAGGAACATCCCTCCTCCTCAGGCCAA CAGCAGCAGGGCCAGGGAGATCGGCAACGGCAGCAAGACCACCATGGTGGACGAGATCCC TATGAGCGTGCCTAGCCTGATGACCGGCCTGAGCCAGGACGACGACTTCGTGCCTTGGCT GAACCACCACCCTAGCCTGGACGGCTACTGCAGCGACTTCCTGAGGGACGTGAGCAGCCC TGTGACCGTGAACGAGCAGGAGAGCGACATGGCCGTGAACCAGACCGCCTTCCCTCTGTT CCAGAGGAGGAAGGACGGCAACGAGAGCGCCCCTGCCGCCAGCAGCAGCCAGTACAACGG CTTCCAGAGCCACAGCCTGTACGGCAGCGACAGGGCCAGGGACCTGCCTAGCCAGCAGAC CAACCCTGACAGGTTCACCCAGACCCAGGAGCCTCTGATCACAAGCAACAAGCCTAGCCT GGTGAACTTCAGCCACTTCCTGAGGCCTGCCACCTTCGCCAAGACCACCAACAACAACCT GCACGACACCAAGGAGAAGAGCCCTCAGAGCCCTCCTAACGTGTTCCAGACCAGGGTGCT GGGCGCCAAGGACAGCGAGGACAAGGTGCTGAACGAGAGCGTGGCCAGCGCCACCCCTAA GGACAACCAGAAGGCCTGCCTGATCAGCGAGGACAGCTGCAGGAAGGACCAGGAGAGCGA GAAGGCTGTGGTGTGCAGCAGCGTGGGCAGCGGCAACAGCCTGGACGGACCTAGCGAGAG CCCTAGCCTGAGCCTGAAGAGGAAGCACAGCAACATCCAGGACATCGACTGCCACAGCGA GGACGTGGAGGAGGAGAGCGGCGACGGAAGGAAGGAGGCTGGACCTAGCAGGACCGGCCT GGGCAGCAAGAGGAGCAGGAGCGCCGAGGTGCACAACCTGTCTGAGAGGAGGAGGAGGGA CAGGATCAACGAGAAGATGAGGGCTCTGCAGGAGCTGATCCCTAACTGCAACAAGGTGGA CAAGGCCAGCATGCTGGACGAGGCCATCGAGTACCTGAAGAGCCTGCAGCTGCAGGTGCA GATCATGAGCATGGCCAGCGGCTACTACCTGCCTCCTGCCGTGATGTTCCCTCCTGGAAT GGGACACTACCCTGCTGCTGCTGCTGCTATGGCTATGGGAATGGGAATGCCTTACGCTAT GGGCCTGCCTGACCTGAGCAGGGGAGGCAGCAGCGTGAACCACGGACCTCAGTTCCAGGT GAGCGGAATGCAGCAGCAGCCTGTGGCTATGGGCATCCCTAGGGTGAGCGGAGGAGGAAT CTTCGCTGGCAGCAGCACCATCGGAAACGGCAGCACCAGGGACCTGAGCGGCAGCAAGGA CCAGACCACCACCAACAACAACAGCAACCTGAAGCCTATCAAGAGGAAGCAGGGCAGCAG CGACCAGTTCTGCGGCAGCAGCTGA
 * 2:Arabidopsis PIF3: Length:1575;

Sequence: ATGAGCGTGAACCTGGCCAGCCAGCTGAGGGAGGGCACCAAGAAGAGCCA CAGCATGGCCGAGAACGTGGGCTTCGTGAAGTGCTTCCTGAAGGGCGTGGTGGAGAAGAA CAGCTACAGGAAGCTGGTGGGCAACCTGTACTTCGTGTACAGCGCCATGGAGGAGGAGAT GGCCAAGTTCAAGGACCACCCTATCCTGAGCCACATCTACTTCCCTGAGCTGAACAGGAA GCAGAGCCTGGAGCAGGACCTGCAGTTCTACTACGGCAGCAACTGGAGGCAGGAGGTGAA GATCAGCGCCGCTGGACAGGCCTACGTGGACAGGGTGAGGCAGGTGGCTGCTACCGCTCC TGAGCTGCTGGTGGCTCACAGCTACACCAGGTACCTGGGCGACCTGAGCGGAGGCCAGAT CCTGAAGAAGATCGCCCAGAACGCCATGAACCTGCACGACGGCGGCACCGCCTTCTACGA GTTCGCCGACATCGACGACGAGAAGGCCTTCAAGAACACCTACAGGCAGGCCATGAACGA CCTGCCTATCGACCAGGCCACCGCCGAGAGGATCGTGGACGAGGCCAACGACGCCTTCGC CATGAACATGAAGATGTTCAACGAGCTGGAGGGCAACCTGATCAAGGCCATCGGCATCAT GGTGTTCAACAGCCTGACCAGGAGGAGGAGCCAGGGCAGCACCGAAGTGGGCCTGGCCAC CAGCGAGGGCTGA
 * 3: Ho1: Length:723;

4.PcyA: Length:747; Sequence: ATGGCCGTGACCGACCTGAGCCTGACCAACAGCAGCCTGATGCCTACCCT GAACCCTATGATCCAGCAGCTGGCCCTGGCCATCGCCGCCAGCTGGCAGAGCCTGCCTCT GAAGCCATACCAGCTGCCTGAGGACCTGGGCTACGTGGAGGGCAGGCTGGAGGGCGAGAA GCTGGTCATCGAGAACAGGTGCTACCAGACCCCTCAGTTCAGGAAGATGCACCTGGAGCT GGCCAAAGTGGGCAAGGGCCTGGACATCCTGCACTGCGTGATGTTCCCTGAGCCTCTGTA CGGCCTGCCTCTGTTCGGATGCGACATCGTGGCTGGACCTGGAGGCGTGAGCGCCGCTAT CGCTGACCTGAGCCCTACCCAGAGCGACAGGCAGCTGCCTGCTGCCTACCAGAAGAGCCT GGCTGAGCTGGGACAGCCTGAGTTCGAGCAGCAGAGGGAGCTGCCTCCTTGGGGAGAGAT CTTCAGCGAGTACTGCCTGTTCATCAGGCCTAGCAACGTGACCGAGGAGGAGAGGTTCGT GCAGAGGGTGGTGGACTTCCTGCAGATCCACTGCCACCAGAGCATCGTGGCCGAGCCTCT GAGCGAGGCCCAGACCCTGGAGCACAGGCAGGGCCAGATCCACTACTGCCAGCAGCAGCA GAAGAACGACAAGACCAGGAGGGTGCTGGAGAAGGCCTTCGGCGAGGCCTGGGCCGAGAG GTACATGAGCCAGGTGCTGTTCGACGTGATCCAGTGA


 * Order plasmid: pCL-CTIG

Wrong！This sequence is repeated. pML1-1F': ttgacgtcaatgggtggagt
 * Design primer for first sequencing:
 * pML1-1F:gtaacaactccgccccatt


 * pML1-1R:tccagctcgaccaggatg


 * Design primer for second sequncing:
 * pML1-1F': ttgacgtcaatgggtggagt
 * pML1-1R2: ACGTCCTCGCCGTCTAAGT

sequencing result

 * GGGACACACGCGCAGACTGTCGACGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCNTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGTAGGGGGCGCGAGGATCCCCCGGGCTGCAGGAATTCCTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGCTCGGTACCCGGGTCGAGGTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTGGAGACGCCATCCACGCTGTTTTGACCTCCATAGAAGACACCGGGACCGATCCAGCCTCCGCGGCCCCGAATTCGATCGTTGGTTTAAACCAACAACCGGTACCTCT-AGCGCTACCGGTCGCCAC


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