GACTTGAAATGGA AGAAAGAAACGTCTGGACCCTCCTGCTCAGGACTTTGTATCTTTGCTAAAGTCAGTGATGTGAAAA TGATATTGCTGATAGGATGAGGATGGATGCTGGAGAAGTGACTTTAGTGAACCACAACTCCGTATTCAAAACCCACCTCCT
下游引物在TGA 后或跨TGA，21bp （三）应用软件：Primer Premier 5
跨ATG和TGA设计引物
TTCTTGACTACTTCTGCGTCTCACGTAAACATTTCTCCAACTCTCCTACTCTGTGGTATCTCCCTGAGATGTGATATCGCTAGTGCCACCATC
TCTGGGAGATCCTCCACCGAAAACTCTTGTGAAGACTGTCTCACTTAGCAGACCTTGACCATGTGGGGGAC
CAGCTCTTTGTTGTCTACAGCCAGAGACCTTGGAAACAGCTGCTCCCAGCCCTCTGCTGTTGTAACACCCTTGATCCTGGACCAGGCCCTGG CGAGATGCATTCACAAGCACATCTGCCTTTCCTTTTGTATCTCAGATACTATTTTTGCAAAGAAACTTTGGTGCTGTGAAAGGGGTGAGGGA CATC
SENP基因查找
https:///
起始密码子ATG，终止密码子TGA，TAA，TAG
酶切位点选用原则
×
1.序列和载体共有酶切位点不选 2.产生甲基化的酶切位点不选 3.要避开用到同尾酶 4.实验室现有酶优先选择
酶切位点
酶切位点名称
HindⅢ KpnI BamHI
AAACCCACCTCCTGCCACAAACAGGTTTTCCAGAGGACCAGCTTTCGCTTTCTGACCAGCAGATTTTATCTTCCAGGCAAGGACATT TGGACCGATCTTTTACATGTTCCACAAGAAGTGCAGCTTATAATCCAAGCTATTACTCAGATAATCCTTCCTCAGACAGTTTTCTTGG CTCAGGCGATTTAAGAACCTTTGGCCAGAGTGCAAATGGCCAATGGAGAAATTCTACCCCATCGTCAAGCTCATCTTTACAAAAATC AAGAAACAGCCGAAGTCTTTACCTCGAAACCCGAAAGACCTCAAGTGGATTATCAAACAGTTTTGCGGGAAAGTCAAACCATCACT GCCATGTATCTGCATATGAAAAATCTTTTCCTATTAAACCTGTTCCAAGTCCATCTTGGAGTGGTTCATGTCGTCGAAGTCTTTTGAG CCCCAAGAAAACTCAGAGGCGACATGTTAGTACAGCAGAAGAGACAGTTCAAGAAGAAGAAAGAGAGATTTACAGACAGCTGCTA CAGATGGTCACAGGGAAACAGTTTACTATAGCCAAACCCACCACACATTTTCCTTTACACCTGTCTCGATGTCTTAGTTCCAGTAAA AATACTTTGAAAGACTCACTGTTTAAAAATGGAAACTCTTGTGCATCTCAGATCATTGGCTCTGATACTTCATCATCTGGATCTGCCA GCATTTTAACTAACCAGGAACAGCTGTCCCACAGTGTATATTCCCTATCTTCTTATACCCCAGATGTTGCATTTGGATCCAAAGATTC TGGTACTCTTCATCATCCCCATCATCACCACTCTGTTCCACATCAGCCAGATAACTTAGCAGCTTCAAATACACAATCTGAAGGATC AGACTCTGTGATTTTACTGAAAGTGAAAGATTCCCAGACTCCAACTCCCAGTTCTACTTTCTTCCAGGCAGAGCTGTGGATCAAAGA ATTAACTAGTGTTTATGATTCTCGAGCACGAGAAAGATTGCGCCAGATTGAAGAACAGAAGGCATTGGCCTTACAGCTTCAAAACC AGAGATTGCAGGAGCGGGAACATTCAGTACATGATTCAGTAGAACTACATCTTCGTGTACCTCTTGAAAAGGAGATTCCTGTTACT GTTGTCCAAGAAACACAAAAAAAAGGTCATAAATTAACTGATAGTGAAGATGAATTTCCTGAAATTACAGAGGAAATGGAGAAAG AAATAAAGAATGTATTTCGTAATGGGAATCAGGATGAAGTTCTCAGTGAAGCATTTCGCCTGACCATTACACGCAAAGATATTCAA ACTCTAAACCATCTGAATTGGCTCAATGATGAGATCATCAATTTCTACATGAATATGCTGATGGAGCGAAGTAAAGAGAAGGGCTT GCCAAGTGTGCATGCATTTAATACCTTTTTCTTCACTAAATTAAAAACGGCTGGTTATCAGGCAGTGAAACGTTGGACAAAGAAAGT AGATGTATTTTCTGTTGACATTCTTTTGGTGCCCATTCACCTGGGAGTACACTGGTGTCTAGCTGTTGTGGACTTTAGAAAGAAGAAT ATTACCTATTACGACTCCATGGGTGGGATAAACAATGAAGCCTGCAGAATACTCTTGCAATACCTAAAGCAAGAAAGCATTGACAA GAAAAGGAAAGAGTTTGACACCAATGGCTGGCAGCTTTTCAGCAAGAAAAGCCAGGAGATTCCTCAGCAGATGAATGGAAGTGAC TGTGGGATGTTTGCCTGCAAATATGCTGACTGTATTACCAAAGACAGACCAATCAACTTCACACAGCAACACATGCCATACTTCCGG
GCCACAAACAGGTTTTCCAGAGGACCAGCTTTCGCTTTCTGACCAGCAGATTTTATCTTCCAGGCAAGGACATTTGGACCGATCTTT TACATGTTCCACAAGAAGTGCAGCTTATAATCCAAGCTATTACTCAGATAATCCTTCCTCAGACAGTTTTCTTGGCTCAGGCGATTT AAGAACCTTTGGCCAGAGTGCAAATGGCCAATGGAGAAATTCTACCCCATCGTCAAGCTCATCTTTACAAAAATCAAGAAACAGCC GAAGTCTTTACCTCGAAACCCGAAAGACCTCAAGTGGATTATCAAACAGTTTTGCGGGAAAGTCAAACCATCACTGCCATGTATCT GCATATGAAAAATCTTTTCCTATTAAACCTGTTCCAAGTCCATCTTGGAGTGGTTCATGTCGTCGAAGTCTTTTGAGCCCCAAGAAA ACTCAGAGGCGACATGTTAGTACAGCAGAAGAGACAGTTCAAGAAGAAGAAAGAGAGATTTACAGACAGCTGCTACAGATGGTCA CAGGGAAACAGTTTACTATAGCCAAACCCACCACACATTTTCCTTTACACCTGTCTCGATGTCTTAGTTCCAGTAAAAATACTTTGA AAGACTCACTGTTTAAAAATGGAAACTCTTGTGCATCTCAGATCATTGGCTCTGATACTTCATCATCTGGATCTGCCAGCATTTTAA CTAACCAGGAACAGCTGTCCCACAGTGTATATTCCCTATCTTCTTATACCCCAGATGTTGCATTTGGATCCAAAGATTCTGGTACTC TTCATCATCCCCATCATCACCACTCTGTTCCACATCAGCCAGATAACTTAGCAGCTTCAAATACACAATCTGAAGGATCAGACTCTG TGATTTTACTGAAAGTGAAAGATTCCCAGACTCCAACTCCCAGTTCTACTTTCTTCCAGGCAGAGCTGTGGATCAAAGAATTAACTA GTGTTTATGATTCTCGAGCACGAGAAAGATTGCGCCAGATTGAAGAACAGAAGGCATTGGCCTTACAGCTTCAAAACCAGAGATTG CAGGAGCGGGAACATTCAGTACATGATTCAGTAGAACTACATCTTCGTGTACCTCTTGAAAAGGAGATTCCTGTTACTGTTGTCCAA GAAACACAAAAAAAAGGTCATAAATTAACTGATAGTGAAGATGAATTTCCTGAAATTACAGAGGAAATGGAGAAAGAAATAAAGAA TGTATTTCGTAATGGGAATCAGGATGAAGTTCTCAGTGAAGCATTTCGCCTGACCATTACACGCAAAGATATTCAAACTCTAAACCA TCTGAATTGGCTCAATGATGAGATCATCAATTTCTACATGAATATGCTGATGGAGCGAAGTAAAGAGAAGGGCTTGCCAAGTGTGC ATGCATTTAATACCTTTTTCTTCACTAAATTAAAAACGGCTGGTTATCAGGCAGTGAAACGTTGGACAAAGAAAGTAGATGTATTTT CTGTTGACATTCTTTTGGTGCCCATTCACCTGGGAGTACACTGGTGTCTAGCTGTTGTGGACTTTAGAAAGAAGAATATTACCTATT ACGACTCCATGGGTGGGATAAACAATGAAGCCTGCAGAATACTCTTGCAATACCTAAAGCAAGAAAGCATTGACAAGAAAAGGAAA GAGTTTGACACCAATGGCTGGCAGCTTTTCAGCAAGAAAAGCCAGGAGATTCCTCAGCAGATGAATGGAAGTGACTGTGGGATGTT TGCCTGCAAATATGCTGACTGTATTACCAAAGACAGACCAATCAACTTCACACAGCAACACATGCCATACTTCCGGAAGCGGATGG