名称: | pCMV-NBN(human)-3×FLAG-EGFP-puro |
---|---|
别称: | NM_002485.5;AT-V1; AT-V2; ATV; NBS; NBS1; P95; |
基因长: | 2262bp |
质粒宿主: | 哺乳细胞 |
质粒用途: | 蛋白表达 |
片段类型: | ORF |
片段物种: | 人 |
原核抗性: | Amp |
筛选标记: | Puro |
荧光标记: | 绿色 |
启动子: | CMV |
复制子: | pUC |
拷贝数: | |
感受态: | DH5a |
温度: | 37度 |
背景载体: | |
正向引物: | CMV-F |
反向引物: | |
诱导方式: |
1、本平台所有产品仅供科研使用,不能用于临床试验,包括人体口服、注射或者体外用药;
2、小包装质粒以模板价格出售,只保证基因部分测序正确,不保证质粒的表达等实验效果;
3、某些质粒即使是出品公司亦没有完整测过序,如关键部位的序列测序正确,亦认为是正确质粒;
4、小包装质粒产品的售后期为您收到产品之日起3个月,收到质粒后请及时转化操作,过期需重新购买;
5、质粒构建皆收取的是成本费用,所有质粒会分享给其他实验者,保密性质粒请提前告知并缴纳保密费用;
6、因科学研究为探索未知的活动,存在较大不确定性,故在任何情况下淼灵都不承担额外的连带责任;
7、发表[中文论文]请标注:产品名称(产品货号) 由武汉淼灵生物科技有限公司提供;
8、发表[英文论文]请标注:Product Name (Product Item Number) was obtained from MiaoLingBio,China.
操作说明
一、质粒干粉(负20度保存,保质期90天,常温运输)
1、收到干粉质粒后请先5000rpm离心1min,再加入20μl无菌水溶解质粒;
2、将相应的感受态于冰盒上解冻,取2μl质粒加至100μl感受态中,冰浴 30min;
3、42度热激60s,冰浴2min,加入900μl无抗LB培养基,180rpm震荡培养45min;
4、6000rpm离心3min,留100μl上清重悬,再涂至对应抗性的LB平板上;
5、倒置培养过夜后,如菌落过多则稀释质粒转化,如没有菌落则取10ul质粒转化;
6、挑单菌落至对应抗性的LB液体培养基中过夜震荡培养,根据需要提取质粒。
二、液体质粒(负20度保存,保质期90天,冰袋运输)
1、收到液体质粒后请甩动保存管,让液体质粒不要吸附在管壁上;
2、根据实验需求取适量质粒,其余保存于负20度冰箱。
三、甘油菌种(负80度保存,保质期90天,冰袋运输)
1、收到甘油菌种后请甩动保存管,让菌液不要吸附在管壁上;
2、用接种环在固体平板上四区划线,如有抗性就加对应抗性;
3、恒温培养箱倒置培养12~16h;
4、务必挑取单菌落接种于液体培养基中,如有抗性就加对应抗性;
5、摇床震荡培养过夜,根据实验需要培养菌种。
CGTTACATAA
CTTACGGTAA
ATGGCCCGCC
TGGCTGACCG
CCCAACGACC
CCCGCCCATT
GACGTCAATA
ATGACGTATG
TTCCCATAGT
AACGCCAATA
GGGACTTTCC
ATTGACGTCA
ATGGGTGGAG
TATTTACGGT
AAACTGCCCA
CTTGGCAGTA
CATCAAGTGT
ATCATATGCC
AAGTACGCCC
CCTATTGACG
TCAATGACGG
TAAATGGCCC
GCCTGGCATT
ATGCCCAGTA
CATGACCTTA
TGGGACTTTC
CTACTTGGCA
GTACATCTAC
GTATTAGTCA
TCGCTATTAC
CATGGTGATG
CGGTTTTGGC
AGTACATCAA
TGGGCGTGGA
TAGCGGTTTG
ACTCACGGGG
ATTTCCAAGT
CTCCACCCCA
TTGACGTCAA
TGGGAGTTTG
TTTTGGCACC
AAAATCAACG
GGACTTTCCA
AAATGTCGTA
ACAACTCCGC
CCCATTGACG
CAAATGGGCG
GTAGGCGTGT
ACGGTGGGAG
GTCTATATAA
GCAGAGCTGG
TTTAGTGAAC
CGTCAGATCC
GCTAGCGCTA
CCGGACTCAG
ATCTCGAGCC
ACCATGTGGA
AACTGCTGCC
CGCCGCGGGC
CCGGCAGGAG
GAGAACCATA
CAGACTTTTG
ACTGGCGTTG
AGTACGTTGT
TGGAAGGAAA
AACTGTGCCA
TTCTGATTGA
AAATGATCAG
TCGATCAGCC
GAAATCATGC
TGTGTTAACT
GCTAACTTTT
CTGTAACCAA
CCTGAGTCAA
ACAGATGAAA
TCCCTGTATT
GACATTAAAA
GATAATTCTA
AGTATGGTAC
CTTTGTTAAT
GAGGAAAAAA
TGCAGAATGG
CTTTTCCCGA
ACTTTGAAGT
CGGGGGATGG
TATTACTTTT
GGAGTGTTTG
GAAGTAAATT
CAGAATAGAG
TATGAGCCTT
TGGTTGCATG
CTCTTCTTGT
TTAGATGTCT
CTGGGAAAAC
TGCTTTAAAT
CAAGCTATAT
TGCAACTTGG
AGGATTTACT
GTAAACAATT
GGACAGAAGA
ATGCACTCAC
CTTGTCATGG
TATCAGTGAA
AGTTACCATT
AAAACAATAT
GTGCACTCAT
TTGTGGACGT
CCAATTGTAA
AGCCAGAATA
TTTTACTGAA
TTCCTGAAAG
CAGTTGAGTC
CAAGAAGCAG
CCTCCACAAA
TTGAAAGTTT
TTACCCACCT
CTTGATGAAC
CATCTATTGG
AAGTAAAAAT
GTTGATCTGT
CAGGACGGCA
GGAAAGAAAA
CAAATCTTCA
AAGGGAAAAC
ATTTATATTT
TTGAATGCCA
AACAGCATAA
GAAATTGAGT
TCCGCAGTTG
TCTTTGGAGG
TGGGGAAGCT
AGGTTGATAA
CAGAAGAGAA
TGAAGAAGAA
CATAATTTCT
TTTTGGCTCC
GGGAACGTGT
GTTGTTGATA
CAGGAATAAC
AAACTCACAG
ACCTTAATTC
CTGACTGTCA
GAAGAAATGG
ATTCAGTCAA
TAATGGATAT
GCTCCAAAGG
CAAGGTCTTA
GACCTATTCC
TGAAGCAGAA
ATTGGATTGG
CGGTGATTTT
CATGACTACA
AAGAATTACT
GTGATCCTCA
GGGCCATCCC
AGTACAGGAT
TAAAGACAAC
AACTCCAGGA
CCAAGCCTTT
CACAAGGCGT
GTCAGTTGAT
GAAAAACTAA
TGCCAAGCGC
CCCAGTGAAC
ACTACAACAT
ACGTAGCTGA
CACAGAATCA
GAGCAAGCAG
ATACATGGGA
TTTGAGTGAA
AGGCCAAAAG
AAATCAAAGT
CTCCAAAATG
GAACAAAAAT
TCAGAATGCT
TTCACAAGAT
GCACCCACTG
TAAAGGAGTC
CTGCAAAACA
AGCTCTAATA
ATAATAGTAT
GGTATCAAAT
ACTTTGGCTA
AGATGAGAAT
CCCAAACTAT
CAGCTTTCAC
CAACTAAATT
GCCAAGTATA
AATAAAAGTA
AAGATAGGGC
TTCTCAGCAG
CAGCAGACCA
ACTCCATCAG
AAACTACTTT
CAGCCGTCTA
CCAAAAAAAG
GGAAAGGGAT
GAAGAAAATC
AAGAAATGTC
TTCATGCAAA
TCAGCAAGAA
TAGAAACGTC
TTGTTCTCTT
TTAGAACAAA
CACAACCTGC
TACACCCTCA
TTGTGGAAAA
ATAAGGAGCA
GCATCTATCT
GAGAATGAGC
CTGTGGACAC
AAACTCAGAC
AATAACTTAT
TTACAGATAC
AGATTTAAAA
TCTATTGTGA
AAAATTCTGC
CAGTAAATCT
CATGCTGCAG
AAAAGCTAAG
ATCAAATAAA
AAAAGGGAAA
TGGATGATGT
GGCCATAGAA
GATGAAGTAT
TGGAACAGTT
ATTCAAGGAC
ACAAAACCAG
AGTTAGAAAT
TGATGTGAAA
GTTCAAAAAC
AGGAGGAAGA
TGTCAATGTT
AGAAAAAGGC
CAAGGATGGA
TATAGAAACA
AATGACACTT
TCAGTGATGA
AGCAGTACCA
GAAAGTAGCA
AAATATCTCA
AGAAAATGAA
ATTGGGAAGA
AACGTGAACT
CAAGGAAGAC
TCACTATGGT
CAGCTAAAGA
AATATCTAAC
AATGACAAAC
TTCAGGATGA
TAGTGAGATG
CTTCCAAAAA
AGCTGTTATT
GACTGAATTT
AGATCACTGG
TGATTAAAAA
CTCTACTTCC
AGAAATCCAT
CTGGCATAAA
TGATGATTAT
GGTCAACTAA
AAAATTTCAA
GAAATTCAAA
AAGGTCACAT
ATCCTGGAGC
AGGAAAACTT
CCACACATCA
TTGGAGGATC
AGATCTAATA
GCTCATCATG
CTCGAAAGAA
TACAGAACTA
GAAGAGTGGC
TAAGGCAGGA
AATGGAGGTA
CAAAATCAAC
ATGCAAAAGA
AGAGTCTCTT
GCTGATGATC
TTTTTAGATA
CAATCCTTAT
TTAAAAAGGA
GAAGAGACTA
CAAAGACCAT
GACGGTGATT
ATAAAGATCA
TGACATCGAC
TACAAGGATG
ACGATGACAA
GTAGTTCGAA
TTCTGCAGTC
GACGGTACCG
CGGGCCCGGG
ATCGATCCGC
CCCTCTCCCT
CCCCCCCCCC
TAACGTTACT
GGCCGAAGCC
GCTTGGAATA
AGGCCGGTGT
GCGTTTGTCT
ATATGTTATT
TTCCACCATA
TTGCCGTCTT
TTGGCAATGT
GAGGGCCCGG
AAACCTGGCC
CTGTCTTCTT
GACGAGCATT
CCTAGGGGTC
TTTCCCCTCT
CGCCAAAGGA
ATGCAAGGTC
TGTTGAATGT
CGTGAAGGAA
GCAGTTCCTC
TGGAAGCTTC
TTGAAGACAA
ACAACGTCTG
TAGCGACCCT
TTGCAGGCAG
CGGAACCCCC
CACCTGGCGA
CAGGTGCCTC
TGCGGCCAAA
AGCCACGTGT
ATAAGATACA
CCTGCAAAGG
CGGCACAACC
CCAGTGCCAC
GTTGTGAGTT
GGATAGTTGT
GGAAAGAGTC
AAATGGCTCT
CCTCAAGCGT
ATTCAACAAG
GGGCTGAAGG
ATGCCCAGAA
GGTACCCCAT
TGTATGGGAT
CTGATCTGGG
GCCTCGGTAC
ACATGCTTTA
CATGTGTTTA
GTCGAGGTTA
AAAAAACGTC
TAGGCCCCCC
GAACCACGGG
GACGTGGTTT
TCCTTTGAAA
AACACGATGA
TAATATGGCC
ACAACCATGG
TGAGCAAGGG
CGAGGAGCTG
TTCACCGGGG
TGGTGCCCAT
CCTGGTCGAG
CTGGACGGCG
ACGTAAACGG
CCACAAGTTC
AGCGTGTCCG
GCGAGGGCGA
GGGCGATGCC
ACCTACGGCA
AGCTGACCCT
GAAGTTCATC
TGCACCACCG
GCAAGCTGCC
CGTGCCCTGG
CCCACCCTCG
TGACCACCCT
GACCTACGGC
GTGCAGTGCT
TCAGCCGCTA
CCCCGACCAC
ATGAAGCAGC
ACGACTTCTT
CAAGTCCGCC
ATGCCCGAAG
GCTACGTCCA
GGAGCGCACC
ATCTTCTTCA
AGGACGACGG
CAACTACAAG
ACCCGCGCCG
AGGTGAAGTT
CGAGGGCGAC
ACCCTGGTGA
ACCGCATCGA
GCTGAAGGGC
ATCGACTTCA
AGGAGGACGG
CAACATCCTG
GGGCACAAGC
TGGAGTACAA
CTACAACAGC
CACAACGTCT
ATATCATGGC
CGACAAGCAG
AAGAACGGCA
TCAAGGTGAA
CTTCAAGATC
CGCCACAACA
TCGAGGACGG
CAGCGTGCAG
CTCGCCGACC
ACTACCAGCA
GAACACCCCC
ATCGGCGACG
GCCCCGTGCT
GCTGCCCGAC
AACCACTACC
TGAGCACCCA
GTCCGCCCTG
AGCAAAGACC
CCAACGAGAA
GCGCGATCAC
ATGGTCCTGC
TGGAGTTCGT
GACCGCCGCC
GGGATCACTC
TCGGCATGGA
CGAGCTGTAC
AAGTCCGGAC
TCAGATCTAG
GAGACGACCT
TCCATGACCG
AGTACAAGCC
CACGGTGCGC
CTCGCCACCC
GCGACGACGT
CCCCCGGGCC
GTACGCACCC
TCGCCGCCGC
GTTCGCCGAC
TACCCCGCCA
CGCGCCACAC
CGTCGACCCG
GACCGCCACA
TCGAGCGGGT
CACCGAGCTG
CAAGAACTCT
TCCTCACGCG
CGTCGGGCTC
GACATCGGCA
AGGTGTGGGT
CGCGGACGAC
GGCGCCGCGG
TGGCGGTCTG
GACCACGCCG
GAGAGCGTCG
AAGCGGGGGC
GGTGTTCGCC
GAGATCGGCC
CGCGCATGGC
CGAGTTGAGC
GGTTCCCGGC
TGGCCGCGCA
GCAACAGATG
GAAGGCCTCC
TGGCGCCGCA
CCGGCCCAAG
GAGCCCGCGT
GGTTCCTGGC
CACCGTCGGC
GTCTCGCCCG
ACCACCAGGG
CAAGGGTCTG
GGCAGCGCCG
TCGTGCTCCC
CGGAGTGGAG
GCGGCCGAGC
GCGCCGGGGT
GCCCGCCTTC
CTGGAGACCT
CCGCGCCCCG
CAACCTCCCC
TTCTACGAGC
GGCTCGGCTT
CACCGTCACC
GCCGACGTCG
AGGTGCCCGA
AGGACCGCGC
ACCTGGTGCA
TGACCCGCAA
GCCCGGTGCG
GGATCCACCG
GATCTAGATA
ACTGATCATA
ATCAGCCATA
CCACATTTGT
AGAGGTTTTA
CTTGCTTTAA
AAAACCTCCC
ACACCTCCCC
CTGAACCTGA
AACATAAAAT
GAATGCAATT
GTTGTTGTTA
ACTTGTTTAT
TGCAGCTTAT
AATGGTTACA
AATAAAGCAA
TAGCATCACA
AATTTCACAA
ATAAAGCATT
TTTTTCACTG
CATTCTAGTT
GTGGTTTGTC
CAAACTCATC
AATGTATCTT
AACGCGATCA
AGCTCCAATT
CGCCCTATAG
TGAGTCGTAT
TACGCGCGCT
CACTGGCCGT
CGTTTTACAA
CGTCGTGACT
GGGAAAACCC
TGGCGTTACC
CAACTTAATC
GCCTTGCAGC
ACATCCCCCT
TTCGCCAGCT
GGCGTAATAG
CGAAGAGGCC
CGCACCGATC
GCCCTTCCCA
ACAGTTGCGC
AGCCTGAATG
GCGAATGGAA
ATTGTAAGCG
TTAATATTTT
GTTAAAATTC
GCGTTAAATT
TTTGTTAAAT
CAGCTCATTT
TTTTAACCAA
TAGGCCGAAA
TCGGCAAAAT
CCCTTATAAA
TCAAAAGAAT
AGACCGAGAT
AGGGTTGAGT
GTTGTTCCAG
TTTGGAACAA
GAGTCCACTA
TTAAAGAACG
TGGACTCCAA
CGTCAAAGGG
CGAAAAACCG
TCTATCAGGG
CGATGGCCCA
CTACGTGAAC
CATCACCCTA
ATCAAGTTTT
TTGGGGTCGA
GGTGCCGTAA
AGCACTAAAT
CGGAACCCTA
AAGGGAGCCC
CCGATTTAGA
GCTTGACGGG
GAAAGCCGGC
GAACGTGGCG
AGAAAGGAAG
GGAAGAAAGC
GAAAGGAGCG
GGCGCTAGGG
CGCTGGCAAG
TGTAGCGGTC
ACGCTGCGCG
TAACCACCAC
ACCCGCCGCG
CTTAATGCGC
CGCTACAGGG
CGCGTCAGGT
GGCACTTTTC
GGGGAAATGT
GCGCGGAACC
CCTATTTGTT
TATTTTTCTA
AATACATTCA
AATATGTATC
CGCTCATGAG
ACAATAACCC
TGATAAATGC
TTCAATAATA
TTGAAAAAGG
AAGAGTATGA
GTATTCAACA
TTTCCGTGTC
GCCCTTATTC
CCTTTTTTGC
GGCATTTTGC
CTTCCTGTTT
TTGCTCACCC
AGAAACGCTG
GTGAAAGTAA
AAGATGCTGA
AGATCAGTTG
GGTGCACGAG
TGGGTTACAT
CGAACTGGAT
CTCAACAGCG
GTAAGATCCT
TGAGAGTTTT
CGCCCCGAAG
AACGTTTTCC
AATGATGAGC
ACTTTTAAAG
TTCTGCTATG
TGGCGCGGTA
TTATCCCGTA
TTGACGCCGG
GCAAGAGCAA
CTCGGTCGCC
GCATACACTA
TTCTCAGAAT
GACTTGGTTG
AGTACTCACC
AGTCACAGAA
AAGCATCTTA
CGGATGGCAT
GACAGTAAGA
GAATTATGCA
GTGCTGCCAT
AACCATGAGT
GATAACACTG
CGGCCAACTT
ACTTCTGACA
ACGATCGGAG
GACCGAAGGA
GCTAACCGCT
TTTTTGCACA
ACATGGGGGA
TCATGTAACT
CGCCTTGATC
GTTGGGAACC
GGAGCTGAAT
GAAGCCATAC
CAAACGACGA
GCGTGACACC
ACGATGCCTG
TAGCAATGGC
AACAACGTTG
CGCAAACTAT
TAACTGGCGA
ACTACTTACT
CTAGCTTCCC
GGCAACAATT
AATAGACTGG
ATGGAGGCGG
ATAAAGTTGC
AGGACCACTT
CTGCGCTCGG
CCCTTCCGGC
TGGCTGGTTT
ATTGCTGATA
AATCTGGAGC
CGGTGAGCGT
GGGTCTCGCG
GTATCATTGC
AGCACTGGGG
CCAGATGGTA
AGCCCTCCCG
TATCGTAGTT
ATCTACACGA
CGGGGAGTCA
GGCAACTATG
GATGAACGAA
ATAGACAGAT
CGCTGAGATA
GGTGCCTCAC
TGATTAAGCA
TTGGTAACTG
TCAGACCAAG
TTTACTCATA
TATACTTTAG
ATTGATTTAA
AACTTCATTT
TTAATTTAAA
AGGATCTAGG
TGAAGATCCT
TTTTGATAAT
CTCATGACCA
AAATCCCTTA
ACGTGAGTTT
TCGTTCCACT
GAGCGTCAGA
CCCCGTAGAA
AAGATCAAAG
GATCTTCTTG
AGATCCTTTT
TTTCTGCGCG
TAATCTGCTG
CTTGCAAACA
AAAAAACCAC
CGCTACCAGC
GGTGGTTTGT
TTGCCGGATC
AAGAGCTACC
AACTCTTTTT
CCGAAGGTAA
CTGGCTTCAG
CAGAGCGCAG
ATACCAAATA
CTGTTCTTCT
AGTGTAGCCG
TAGTTAGGCC
ACCACTTCAA
GAACTCTGTA
GCACCGCCTA
CATACCTCGC
TCTGCTAATC
CTGTTACCAG
TGGCTGCTGC
CAGTGGCGAT
AAGTCGTGTC
TTACCGGGTT
GGACTCAAGA
CGATAGTTAC
CGGATAAGGC
GCAGCGGTCG
GGCTGAACGG
GGGGTTCGTG
CACACAGCCC
AGCTTGGAGC
GAACGACCTA
CACCGAACTG
AGATACCTAC
AGCGTGAGCT
ATGAGAAAGC
GCCACGCTTC
CCGAAGGGAG
AAAGGCGGAC
AGGTATCCGG
TAAGCGGCAG
GGTCGGAACA
GGAGAGCGCA
CGAGGGAGCT
TCCAGGGGGA
AACGCCTGGT
ATCTTTATAG
TCCTGTCGGG
TTTCGCCACC
TCTGACTTGA
GCGTCGATTT
TTGTGATGCT
CGTCAGGGGG
GCGGAGCCTA
TGGAAAAACG
CCAGCAACGC
GGCCTTTTTA
CGGTTCCTGG
CCTTTTGCTG
GCCTTTTGCT
CACATGTTCT
TTCCTGCGTT
ATCCCCTGAT
TCTGTGGATA
ACCGTATTAC
CGCCTTTGAG
TGAGCTGATA
CCGCTCGCCG
CAGCCGAACG
ACCGAGCGCA
GCGAGTCAGT
GAGCGAGGAA
GCGGAAGAGC
GCCCAATACG
CAAACCGCCT
CTCCCCGCGC
GTTGGCCGAT
TCATTAATGC
AGCTGGCACG
ACAGGTTTCC
CGACTGGAAA
GCGGGCAGTG
AGCGCAACGC
AATTAATGTG
AGTTAGCTCA
CTCATTAGGC
ACCCCAGGCT
TTACACTTTA
TGCTTCCGGC
TCGTATGTTG
TGTGGAATTG
TGAGCGGATA
ACAATTTCAC
ACAGGAAACA
GCTATGACCA
TGATTACGCC
AAGCGCGCAA
TTAACCCTCA
CTAAAGGGAA
CAAAAGCTGG
CTAGTGGATC
CCCCGGGCTG
CAGGAATTCG
ATTAATAGTA
ATCAATTACG
GGGTCATTAG
TTCATAGCCC
ATATATGGAG
TTCCG