人源MEIOB-CopGFP/SPATA22-mCherry哺乳动物融合表达质粒

• 质粒名称：

  pcDNA3.1-CMV-human_MEIOB_CopGFP-CMV-human_SPATA22_mCherry

• 载体骨架：

  pcDNA3.1

• 载体骨架片段大小（bp）：

  4552

• 载体总大小（bp）：

  8846

• 质粒类型：

  哺乳动物荧光表达质粒

• 骨架修饰：

  本载体为pcDNA3.1的改造版本，插入人源MEIOB基因，并新增了CopGFP标签；插入人源SPATA22基因，并新增了mCherry标签，适用于双荧光标记实验；插入转座酶识别位点，可用于构建稳转细胞系；插入嘌呤霉素抗性基因用于筛选。

• 质粒储存：

  -80°C（长期）或-20°C（6个月内）。

• 质粒浓度：

  1 μg/μl，溶于无菌水。

• 质粒抗性：

  质粒携带AmpR基因，培养时添加70-100 μg/mL氨苄青霉素。

• 培养条件：

  将DH5α/pcDNA3.1菌株接种于含70-100 μg/mL氨苄青霉素的LB液体培养基，37°C、200 rpm振荡培养。

• 克隆用感受态：

  DH5α

• 表达用感受态：

  无

• 高拷贝数/低拷贝数：

  500-700

• 基因/插入片段名称：

  hMEIOB；hSPATA22

• 质粒来源：

  自行构建

• 突变情况：

  密码子优化（序列适合原核表达）；GC序列优化（40-60%）。

• 启动子：

  human cytomegalovirus (CMV) immediate early promoter

• 标签/融合蛋白：

  CopGFP；mCherry

载体序列：gcactctcagtacaatctgctctgatgccgcatagttaagccagtatctgctccctgcttgtgtgttggaggtcttaacccgctgagtagtgcgcgagcaaaatttaagctacaacaaggcaaggcttgaccgacaattgcatgaagaatctgcttagggttaggcgttttgcgctgcttcgcgatgtacgggccagatatacgcgttgacattgattattgactagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccgcgttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaaactgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctctctggctaactagagaacccactgcttactggcttatcgaaattaatacgactcactatagggagacccaagctggctagcatggcgaacagcttcgcagcacgtatttttactactctgagcgatctgcagaccaacatggcgaacctgaaggtgattggcattgtgattggcaaaaccgatgtgaaaggctttccggatcgcaaaaacattggcagcgaacgctatacctttagctttaccattcgcgatagtccagcacattttgttaatgcagcaagctggggcaacgaggactacatcaaaagtctgagcgatagctttcgcgtgggcgattgcgtgattattgaaaacccgctgattcagcgcaaggagatcgaacgtgaagaaaagtttagtccagcaactccgagcaactgcaaactgctgctgagcgaaaaccatagcaccgtgaaagtgtgcagcagctatgaagtggataccaaactgctgagcctgattcacctgccggtgaaggaaagccatgactactacagcctgggcgatattgttgcgaatggtcacagcctgaacggccgtatcatcaatgttttagcagcggtgaaaagcgtgggcgaaccgaagtactttaccaccagtgatcgtcgtaaaggtcaacgctgcgaagtgcgcctgtatgatgaaaccgaaagcagctttgcgatgacctgctgggacaacgagagcatcttattagcacaaagttggatgccacgtgaaaccgtgatcttcgcgagcgatgtgcgcatcaacttcgacaagtttcgcaactgcatgaccgcgaccgtgattagcaaaaccattattaccaccaacccggatattccggaagcgaacattctgctgaacttcattcgcgagaacaaggaaaccaacgtgctggacgacgagatcgacagctacttcaaggaaagcattaacctgagcaccattgtggatgtgtataccgtggaacagctgaaaggcaaagcgctgaaaaacgaaggcaaagcggatccgagctatggcattctgtatgcgtacatcagcaccctgaacatcgatgatgaaaccaccaaagttgttcgtaaccgctgcagcagctgcggctatattgtgaacgaagcgagcaacatgtgcaccacctgcaacaagaacagcctggacttcaagagcgtgtttctgagctttcatgtgctgattgatctgaccgatcatactggtactctgcatagttgtagcctgactggtagtgttgcagaagaaaccctgggctgcaccgtgcatgaatttctggcgatgaccgatgaacagaaaaccgcgctgaaatggcagttcctgctggagcgcagcaagatctacctgaagttcgttctgagtcatcgtgcacgtagcggcctgaaaattagcgtgctgagttgtaaactggcagatccaactgaagcgagtcgtaacctgagtggtcagaaacatgtgTtActcgagatggagagcgacgagagcggcctgcccgccatggagatcgagtgccgcatcaccggcaccctgaacggcgtggagttcgagctggtgggcggcggagagggcacccccaagcagggccgcatgaccaacaagatgaagagcaccaaaggcgccctgaccttcagcccctacctgctgagccacgtgatgggctacggcttctaccacttcggcacctaccccagcggctacgagaaccccttcctgcacgccatcaacaacggcggctacaccaacacccgcatcgagaagtacgaggacggcggcgtgctgcacgtgagcttcagctaccgctacgaggccggccgcgtgatcggcgacttcaaggtggtgggcaccggcttccccgaggacagcgtgatcttcaccgacaagatcatccgcagcaacgccaccgtggagcacctgcaccccatgggcgataacgtgctggtgggcagcttcgcccgcaccttcagcctgcgcgacggcggctactacagcttcgtggtggacagccacatgcacttcaagagcgccatccaccccagcatcctgcagaacgggggccccatgttcgccttccgccgcgtggaggagctgcacagcaacaccgagctgggcatcgtggagtaccagcacgccttcaagacccccatcgccttcgccagatcccgcgctcagtcgtccaattctgccgtggacggcaccgccggacccggctccaccggatctcgctaagtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctctctggctaactagagaacccactgcttactggcttatcgaaattaatacgactcactatagggagacccaagctggctagcatgaaacgtagtctgaacgaaaatagtgcacgtagtactgcaggttgcctgccagttccactgttgaaccagaagaagcgcaaccgtcaaccactgaccagcaacccactgaaagatgatagcggcattagcaccccgagcgataactatgattttccaccactgccaactgattgggcatgggaagcagttaatccagaactggcaccagtgatgaagaccgttgatactggtcagattccacatagtgtgagtcgtccactgcgtagtcaggacagtgtgttcaacagcattcagagtaatactggtcgtagtcaaggtggttggagttatcgtgatggcaacaagaacaccagcctgaagacctggaacaagaacgactttaaaccgcaatgtaagcgcaccaacctggttgcgaacgacggcaagaatagttgtccagttagtagtggtgcacaacagcaaaaacagttacgtattccagaaccaccaaatctgagccgcaacaaagaaaccgaactgctgcgccagacccatagtagtaagattagtggttgcactatgcgtggtctggataaaaacagcgcgctgcagaccctgaagccgaacttccagcagaaccagtacaagaagcagatgctggatgatattccggaagataacaccctgaaagaaaccagcctgtatcagctgcagtttaaagaaaaagcgagcagcctgcgcattattagcgcggtgattgaaagcatgaaatattggcgcgaacatgcgcagaaaaccgtgctgctgtttgaagtgttagcggtgctggatagtgcagttaccccaggcccatattatagcaaaacctttctgatgcgcgatggcaagaacaccctgccgtgcgtgttctacgagatcgatcgtgaattaccacgcttaattcgtggtcgtgtgcatcgctgcgtgggcaactacgaccagaagaagaacatcttccagtgtgttagtgttcgtccagcaagtgttagcgaacagaaaacctttcaggcgttcgtgaagattgcggatgtggagatgcagtactatattaacgtgatgaacgaaactggTggcggTggTagtggCggcggTggTagtATGGTGAGCAAGGGCGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGGGCTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACATCCTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTGCAGGACGGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAGACCATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGAGATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAGGTCAAGACCACCTACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGACATCACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAACGCGCCGAGGGCCGCCACTCCACCGGCGGCATGGACGAGCTGTACAAGTAAgtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctctctggctaactagagaacccactgcGCCGCCatgaccgagtacaagcccacggtgcgcctcgccacccgcgacgacgtccccagggccgtacgcaccctcgccgccgcgttcgccgactaccccgccacgcgccacaccgtcgatccggaccgccacatcgagcgggtcaccgagctgcaagaactcttcctcacgcgcgtcgggctcgacatcggcaaggtgtgggtcgcggacgacggcgccgcggtggcggtctggaccacgccggagagcgtcgaagcgggggcggtgttcgccgagatcggcccgcgcatggccgagttgagcggttcccggctggccgcgcagcaacagatggaaggcctcctggcgccgcaccggcccaaggagcccgcgtggttcctggccaccgtcggcgtctcgcccgaccaccagggcaagggtctgggcagcgccgtcgtgctccccggagtggaggcggccgagcgcgccggggtgcccgccttcctggagacctccgcgccccgcaacctccccttctacgagcggctcggcttcaccgtcaccgccgacgtcgaggtgcccgaaggaccgcgcacctggtgcatgacccgcaagcccggtgcctgagatcagcctcgactgtgccttctagttgccagccatctgttgtttgcccctcccccgtgccttccttgaccctggaaggtgccactcccactgtcctttcctaataaaatgaggaaattgcatcgcattgtctgagtaggtgtcattctattctggggggtggggtggggcaggacagcaagggggaggattgggaagacaatagcaggcatgctggggatgcggtgggctctatggcttctgaggcggaaagaaccagctggggctctagggggtatccccacgcgccctgtagcggcgcattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcgcccgctcctttcgctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctccctttagggttccgatttagtgctttacggcacctcgaccccaaaaaacttgattagggtgatggttcacgtagtgggccatcgccctgatagacggtttttcgccctttgacgttggagtccacgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatctcggtctattcttttgatttataagggattttgccgatttcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaattaattctgtggaatcctggggtgcctaatgagtgagctaactcacattaattgcgttgcgctcactgcccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggtttgcgtattgggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccgggttaaacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcacatttccccgaaaagtgccacctgacgtcgacggatcgggagatctcccgatcccctatggt