名称: | pLV2-CMV-3×FLAG-Map1lc3b(mouse)-Puro |
---|---|
别称: | NM_026160.5;1010001C15Rik; Atg8; LC3b; MAP1A/MAP1B;Map1lc3 |
基因长: | 378bp |
质粒宿主: | 哺乳细胞,慢病毒 |
质粒用途: | 蛋白表达 |
片段类型: | ORF |
片段物种: | 小鼠 |
原核抗性: | Amp |
筛选标记: | Puro |
荧光标记: | |
启动子: | CMV |
复制子: | pUC |
拷贝数: | |
感受态: | Stbl3 |
温度: | 37度 |
背景载体: | pLV2-CMV-3×FLAG-MCS-Puro |
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.
pLV2-CMV-3×FLAG-Map1lc3b(mouse)-Puro是适用于哺乳细胞过表达的质粒。质粒带有的CMV强启动子可启动小鼠源Map1lc3b基因转录,促进Map1lc3b基因N端融合表达3×FLAG标签,便于FLAG抗体对表达的融合蛋白进行检测、定位和纯化等。此外,质粒带有HIV-1 Ψ和3'/5'LTR元件可用于慢病毒包装,便于基因稳定整合至宿主细胞。质粒携带Puro抗性基因,用于筛选表达目的基因的宿主细胞。
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。
TGGAAGGGCT
AATTCACTCC
CAAAGAAGAC
AAGATATCCT
TGATCTGTGG
ATCTACCACA
CACAAGGCTA
CTTCCCTGAT
TAGCAGAACT
ACACACCAGG
GCCAGGGGTC
AGATATCCAC
TGACCTTTGG
ATGGTGCTAC
AAGCTAGTAC
CAGTTGAGCC
AGATAAGGTA
GAAGAGGCCA
ATAAAGGAGA
GAACACCAGC
TTGTTACACC
CTGTGAGCCT
GCATGGGATG
GATGACCCGG
AGAGAGAAGT
GTTAGAGTGG
AGGTTTGACA
GCCGCCTAGC
ATTTCATCAC
GTGGCCCGAG
AGCTGCATCC
GGAGTACTTC
AAGAACTGCT
GATATCGAGC
TTGCTACAAG
GGACTTTCCG
CTGGGGACTT
TCCAGGGAGG
CGTGGCCTGG
GCGGGACTGG
GGAGTGGCGA
GCCCTCAGAT
CCTGCATATA
AGCAGCTGCT
TTTTGCCTGT
ACTGGGTCTC
TCTGGTTAGA
CCAGATCTGA
GCCTGGGAGC
TCTCTGGCTA
ACTAGGGAAC
CCACTGCTTA
AGCCTCAATA
AAGCTTGCCT
TGAGTGCTTC
AAGTAGTGTG
TGCCCGTCTG
TTGTGTGACT
CTGGTAACTA
GAGATCCCTC
AGACCCTTTT
AGTCAGTGTG
GAAAATCTCT
AGCAGTGGCG
CCCGAACAGG
GACTTGAAAG
CGAAAGGGAA
ACCAGAGGAG
CTCTCTCGAC
GCAGGACTCG
GCTTGCTGAA
GCGCGCACGG
CAAGAGGCGA
GGGGCGGCGA
CTGGTGAGTA
CGCCAAAAAT
TTTGACTAGC
GGAGGCTAGA
AGGAGAGAGA
TGGGTGCGAG
AGCGTCAGTA
TTAAGCGGGG
GAGAATTAGA
TCGCGATGGG
AAAAAATTCG
GTTAAGGCCA
GGGGGAAAGA
AAAAATATAA
ATTAAAACAT
ATAGTATGGG
CAAGCAGGGA
GCTAGAACGA
TTCGCAGTTA
ATCCTGGCCT
GTTAGAAACA
TCAGAAGGCT
GTAGACAAAT
ACTGGGACAG
CTACAACCAT
CCCTTCAGAC
AGGATCAGAA
GAACTTAGAT
CATTATATAA
TACAGTAGCA
ACCCTCTATT
GTGTGCATCA
AAGGATAGAG
ATAAAAGACA
CCAAGGAAGC
TTTAGACAAG
ATAGAGGAAG
AGCAAAACAA
AAGTAAGACC
ACCGCACAGC
AAGCGGCCGG
CCGCTGATCT
TCAGACCTGG
AGGAGGAGAT
ATGAGGGACA
ATTGGAGAAG
TGAATTATAT
AAATATAAAG
TAGTAAAAAT
TGAACCATTA
GGAGTAGCAC
CCACCAAGGC
AAAGAGAAGA
GTGGTGCAGA
GAGAAAAAAG
AGCAGTGGGA
ATAGGAGCTT
TGTTCCTTGG
GTTCTTGGGA
GCAGCAGGAA
GCACTATGGG
CGCAGCGTCA
ATGACGCTGA
CGGTACAGGC
CAGACAATTA
TTGTCTGGTA
TAGTGCAGCA
GCAGAACAAT
TTGCTGAGGG
CTATTGAGGC
GCAACAGCAT
CTGTTGCAAC
TCACAGTCTG
GGGCATCAAG
CAGCTCCAGG
CAAGAATCCT
GGCTGTGGAA
AGATACCTAA
AGGATCAACA
GCTCCTGGGG
ATTTGGGGTT
GCTCTGGAAA
ACTCATTTGC
ACCACTGCTG
TGCCTTGGAA
TGCTAGTTGG
AGTAATAAAT
CTCTGGAACA
GATTTGGAAT
CACACGACCT
GGATGGAGTG
GGACAGAGAA
ATTAACAATT
ACACAAGCTT
AATACACTCC
TTAATTGAAG
AATCGCAAAA
CCAGCAAGAA
AAGAATGAAC
AAGAATTATT
GGAATTAGAT
AAATGGGCAA
GTTTGTGGAA
TTGGTTTAAC
ATAACAAATT
GGCTGTGGTA
TATAAAATTA
TTCATAATGA
TAGTAGGAGG
CTTGGTAGGT
TTAAGAATAG
TTTTTGCTGT
ACTTTCTATA
GTGAATAGAG
TTAGGCAGGG
ATATTCACCA
TTATCGTTTC
AGACCCACCT
CCCAACCCCG
AGGGGACCCG
ACAGGCCCGA
AGGAATAGAA
GAAGAAGGTG
GAGAGAGAGA
CAGAGACAGA
TCCATTCGAT
TAGTGAACGG
ATCTCGACGG
TATCGCCTTT
AAAAGAAAAG
GGGGGATTGG
GGGGTACAGT
GCAGGGGAAA
GAATAGTAGA
CATAATAGCA
ACAGACATAC
AAACTAAAGA
ATTACAAAAA
CAAATTACAA
AAATTCAAAA
TTTTCGGGTT
TATTACAGGG
ACAGCAGAGA
TCCAGTTTAT
CGATAAGCTT
GGGAGTTCCG
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
GCAGAGCTCG
TTTAGTGAAC
CGTCAGATCG
CCTGGAGACG
CCATCCACGC
TGTTTTGACC
TCCATAGAAG
ACACCGACTC
TACTAGAGGA
TCGCTAGCGC
TACCGGACTC
AGATCTCGAG
CCACCATGGA
CTACAAAGAC
CATGACGGTG
ATTATAAAGA
TCATGACATC
GACTACAAGG
ATGACGATGA
CAAGGAATTC
ATGCCGTCCG
AGAAGACCTT
CAAGCAGCGC
CGGAGCTTTG
AACAAAGAGT
GGAAGATGTC
CGGCTCATCC
GGGAGCAGCA
CCCCACCAAG
ATCCCAGTGA
TTATAGAGCG
ATACAAGGGG
GAGAAGCAGC
TGCCCGTCCT
GGACAAGACC
AAGTTCCTGG
TGCCTGACCA
CGTGAACATG
AGCGAGCTCA
TCAAGATAAT
CAGACGGCGC
TTGCAGCTCA
ATGCTAACCA
AGCCTTCTTC
CTCCTGGTGA
ATGGGCACAG
CATGGTGAGT
GTGTCCACTC
CCATCTCCGA
AGTGTACGAG
AGTGAGAGAG
ATGAAGACGG
CTTCCTGTAC
ATGGTTTATG
CCTCGCAGGA
GACATTCGGG
ACAGCAATGG
CTGTGTAAGG
ATCCCGCGAC
TCTAGATAAT
TCTACCGGGT
AGGGGAGGCG
CTTTTCCCAA
GGCAGTCTGG
AGCATGCGCT
TTAGCAGCCC
CGCTGGGCAC
TTGGCGCTAC
ACAAGTGGCC
TCTGGCCTCG
CACACATTCC
ACATCCACCG
GTAGGCGCCA
ACCGGCTCCG
TTCTTTGGTG
GCCCCTTCGC
GCCACCTTCT
ACTCCTCCCC
TAGTCAGGAA
GTTCCCCCCC
GCCCCGCAGC
TCGCGTCGTG
CAGGACGTGA
CAAATGGAAG
TAGCACGTCT
CACTAGTCTC
GTGCAGATGG
ACAGCACCGC
TGAGCAATGG
AAGCGGGTAG
GCCTTTGGGG
CAGCGGCCAA
TAGCAGCTTT
GCTCCTTCGC
TTTCTGGGCT
CAGAGGCTGG
GAAGGGGTGG
GTCCGGGGGC
GGGCTCAGGG
GCGGGCTCAG
GGGCGGGGCG
GGCGCCCGAA
GGTCCTCCGG
AGGCCCGGCA
TTCTGCACGC
TTCAAAAGCG
CACGTCTGCC
GCGCTGTTCT
CCTCTTCCTC
ATCTCCGGGC
CTTTCGACCT
GCAGCCCAAG
CTTACCATGA
CCGAGTACAA
GCCCACGGTG
CGCCTCGCCA
CCCGCGACGA
CGTCCCCAGG
GCCGTACGCA
CCCTCGCCGC
CGCGTTCGCC
GACTACCCCG
CCACGCGCCA
CACCGTCGAT
CCGGACCGCC
ACATCGAGCG
GGTCACCGAG
CTGCAAGAAC
TCTTCCTCAC
GCGCGTCGGG
CTCGACATCG
GCAAGGTGTG
GGTCGCGGAC
GACGGCGCCG
CGGTGGCGGT
CTGGACCACG
CCGGAGAGCG
TCGAAGCGGG
GGCGGTGTTC
GCCGAGATCG
GCCCGCGCAT
GGCCGAGTTG
AGCGGTTCCC
GGCTGGCCGC
GCAGCAACAG
ATGGAAGGCC
TCCTGGCGCC
GCACCGGCCC
AAGGAGCCCG
CGTGGTTCCT
GGCCACCGTC
GGCGTCTCGC
CCGACCACCA
GGGCAAGGGT
CTGGGCAGCG
CCGTCGTGCT
CCCCGGAGTG
GAGGCGGCCG
AGCGCGCCGG
GGTGCCCGCC
TTCCTGGAGA
CCTCCGCGCC
CCGCAACCTC
CCCTTCTACG
AGCGGCTCGG
CTTCACCGTC
ACCGCCGACG
TCGAGGTGCC
CGAAGGACCG
CGCACCTGGT
GCATGACCCG
CAAGCCCGGT
GCCTGACCGC
GTCTGGAACA
ATCAACCTCT
GGATTACAAA
ATTTGTGAAA
GATTGACTGG
TATTCTTAAC
TATGTTGCTC
CTTTTACGCT
ATGTGGATAC
GCTGCTTTAA
TGCCTTTGTA
TCATGCTATT
GCTTCCCGTA
TGGCTTTCAT
TTTCTCCTCC
TTGTATAAAT
CCTGGTTGCT
GTCTCTTTAT
GAGGAGTTGT
GGCCCGTTGT
CAGGCAACGT
GGCGTGGTGT
GCACTGTGTT
TGCTGACGCA
ACCCCCACTG
GTTGGGGCAT
TGCCACCACC
TGTCAGCTCC
TTTCCGGGAC
TTTCGCTTTC
CCCCTCCCTA
TTGCCACGGC
GGAACTCATC
GCCGCCTGCC
TTGCCCGCTG
CTGGACAGGG
GCTCGGCTGT
TGGGCACTGA
CAATTCCGTG
GTGTTGTCGG
GGAAGCTGAC
GTCCTTTCCA
TGGCTGCTCG
CCTGTGTTGC
CACCTGGATT
CTGCGCGGGA
CGTCCTTCTG
CTACGTCCCT
TCGGCCCTCA
ATCCAGCGGA
CCTTCCTTCC
CGCGGCCTGC
TGCCGGCTCT
GCGGCCTCTT
CCGCGTCTTC
GCCTTCGCCC
TCAGACGAGT
CGGATCTCCC
TTTGGGCCGC
CTCCCCGCCT
GGAATTAATT
CTGCAGTCGA
GACCTAGAAA
AACATGGAGC
AATCACAAGT
AGCAATACAG
CAGCTACCAA
TGCTGATTGT
GCCTGGCTAG
AAGCACAAGA
GGAGGAGGAG
GTGGGTTTTC
CAGTCACACC
TCAGGTACCT
TTAAGACCAA
TGACTTACAA
GGCAGCTGTA
GATCTTAGCC
ACTTTTTAAA
AGAAAAGAGG
GGACTGGAAG
GGCTAATTCA
CTCCCAACGA
AGACAAGATA
TCCTTGATCT
GTGGATCTAC
CACACACAAG
GCTACTTCCC
TGATTAGCAG
AACTACACAC
CAGGGCCAGG
GGTCAGATAT
CCACTGACCT
TTGGATGGTG
CTACAAGCTA
GTACCAGTTG
AGCCAGATAA
GGTAGAAGAG
GCCAATAAAG
GAGAGAACAC
CAGCTTGTTA
CACCCTGTGA
GCCTGCATGG
GATGGATGAC
CCGGAGAGAG
AAGTGTTAGA
GTGGAGGTTT
GACAGCCGCC
TAGCATTTCA
TCACGTGGCC
CGAGAGCTGC
ATCCGGAGTA
CTTCAAGAAC
TGCTGATATC
GAGCTTGCTA
CAAGGGACTT
TCCGCTGGGG
ACTTTCCAGG
GAGGCGTGGC
CTGGGCGGGA
CTGGGGAGTG
GCGAGCCCTC
AGATCCTGCA
TATAAGCAGC
TGCTTTTTGC
CTGTACTGGG
TCTCTCTGGT
TAGACCAGAT
CTGAGCCTGG
GAGCTCTCTG
GCTAACTAGG
GAACCCACTG
CTTAAGCCTC
AATAAAGCTT
GCCTTGAGTG
CTTCAAGTAG
TGTGTGCCCG
TCTGTTGTGT
GACTCTGGTA
ACTAGAGATC
CCTCAGACCC
TTTTAGTCAG
TGTGGAAAAT
CTCTAGCAGT
AGTAGTTCAT
GTCATCTTAT
TATTCAGTAT
TTATAACTTG
CAAAGAAATG
AATATCAGAG
AGTGAGAGGC
CTTGACATTG
CTAGCGTTTT
ACCGTCGACC
TCTAGCTAGA
GCTTGGCGTA
ATCATGGTCA
TAGCTGTTTC
CTGTGTGAAA
TTGTTATCCG
CTCACAATTC
CACACAACAT
ACGAGCCGGA
AGCATAAAGT
GTAAAGCCTG
GGGTGCCTAA
TGAGTGAGCT
AACTCACATT
AATTGCGTTG
CGCTCACTGC
CCGCTTTCCA
GTCGGGAAAC
CTGTCGTGCC
AGCTGCATTA
ATGAATCGGC
CAACGCGCGG
GGAGAGGCGG
TTTGCGTATT
GGGCGCTCTT
CCGCTTCCTC
GCTCACTGAC
TCGCTGCGCT
CGGTCGTTCG
GCTGCGGCGA
GCGGTATCAG
CTCACTCAAA
GGCGGTAATA
CGGTTATCCA
CAGAATCAGG
GGATAACGCA
GGAAAGAACA
TGTGAGCAAA
AGGCCAGCAA
AAGGCCAGGA
ACCGTAAAAA
GGCCGCGTTG
CTGGCGTTTT
TCCATAGGCT
CCGCCCCCCT
GACGAGCATC
ACAAAAATCG
ACGCTCAAGT
CAGAGGTGGC
GAAACCCGAC
AGGACTATAA
AGATACCAGG
CGTTTCCCCC
TGGAAGCTCC
CTCGTGCGCT
CTCCTGTTCC
GACCCTGCCG
CTTACCGGAT
ACCTGTCCGC
CTTTCTCCCT
TCGGGAAGCG
TGGCGCTTTC
TCATAGCTCA
CGCTGTAGGT
ATCTCAGTTC
GGTGTAGGTC
GTTCGCTCCA
AGCTGGGCTG
TGTGCACGAA
CCCCCCGTTC
AGCCCGACCG
CTGCGCCTTA
TCCGGTAACT
ATCGTCTTGA
GTCCAACCCG
GTAAGACACG
ACTTATCGCC
ACTGGCAGCA
GCCACTGGTA
ACAGGATTAG
CAGAGCGAGG
TATGTAGGCG
GTGCTACAGA
GTTCTTGAAG
TGGTGGCCTA
ACTACGGCTA
CACTAGAAGA
ACAGTATTTG
GTATCTGCGC
TCTGCTGAAG
CCAGTTACCT
TCGGAAAAAG
AGTTGGTAGC
TCTTGATCCG
GCAAACAAAC
CACCGCTGGT
AGCGGTGGTT
TTTTTGTTTG
CAAGCAGCAG
ATTACGCGCA
GAAAAAAAGG
ATCTCAAGAA
GATCCTTTGA
TCTTTTCTAC
GGGGTCTGAC
GCTCAGTGGA
ACGAAAACTC
ACGTTAAGGG
ATTTTGGTCA
TGAGATTATC
AAAAAGGATC
TTCACCTAGA
TCCTTTTAAA
TTAAAAATGA
AGTTTTAAAT
CAATCTAAAG
TATATATGAG
TAAACTTGGT
CTGACAGTTA
CCAATGCTTA
ATCAGTGAGG
CACCTATCTC
AGCGATCTGT
CTATTTCGTT
CATCCATAGT
TGCCTGACTC
CCCGTCGTGT
AGATAACTAC
GATACGGGAG
GGCTTACCAT
CTGGCCCCAG
TGCTGCAATG
ATACCGCGAG
ACCCACGCTC
ACCGGCTCCA
GATTTATCAG
CAATAAACCA
GCCAGCCGGA
AGGGCCGAGC
GCAGAAGTGG
TCCTGCAACT
TTATCCGCCT
CCATCCAGTC
TATTAATTGT
TGCCGGGAAG
CTAGAGTAAG
TAGTTCGCCA
GTTAATAGTT
TGCGCAACGT
TGTTGCCATT
GCTACAGGCA
TCGTGGTGTC
ACGCTCGTCG
TTTGGTATGG
CTTCATTCAG
CTCCGGTTCC
CAACGATCAA
GGCGAGTTAC
ATGATCCCCC
ATGTTGTGCA
AAAAAGCGGT
TAGCTCCTTC
GGTCCTCCGA
TCGTTGTCAG
AAGTAAGTTG
GCCGCAGTGT
TATCACTCAT
GGTTATGGCA
GCACTGCATA
ATTCTCTTAC
TGTCATGCCA
TCCGTAAGAT
GCTTTTCTGT
GACTGGTGAG
TACTCAACCA
AGTCATTCTG
AGAATAGTGT
ATGCGGCGAC
CGAGTTGCTC
TTGCCCGGCG
TCAATACGGG
ATAATACCGC
GCCACATAGC
AGAACTTTAA
AAGTGCTCAT
CATTGGAAAA
CGTTCTTCGG
GGCGAAAACT
CTCAAGGATC
TTACCGCTGT
TGAGATCCAG
TTCGATGTAA
CCCACTCGTG
CACCCAACTG
ATCTTCAGCA
TCTTTTACTT
TCACCAGCGT
TTCTGGGTGA
GCAAAAACAG
GAAGGCAAAA
TGCCGCAAAA
AAGGGAATAA
GGGCGACACG
GAAATGTTGA
ATACTCATAC
TCTTCCTTTT
TCAATATTAT
TGAAGCATTT
ATCAGGGTTA
TTGTCTCATG
AGCGGATACA
TATTTGAATG
TATTTAGAAA
AATAAACAAA
TAGGGGTTCC
GCGCACATTT
CCCCGAAAAG
TGCCACCTGA
CGTCGACGGA
TCGGGAGATC
AACTTGTTTA
TTGCAGCTTA
TAATGGTTAC
AAATAAAGCA
ATAGCATCAC
AAATTTCACA
AATAAAGCAT
TTTTTTCACT
GCATTCTAGT
TGTGGTTTGT
CCAAACTCAT
CAATGTATCT
TATCATGTCT
GGATCAACTG
GATAACTCAA
GCTAACCAAA
ATCATCCCAA
ACTTCCCACC
CCATACCCTA
TTACCACTGC
CAATTACCTG
TGGTTTCATT
TACTCTAAAC
CTGTGATTCC
TCTGAATTAT
TTTCATTTTA
AAGAAATTGT
ATTTGTTAAA
TATGTACTAC
AAACTTAGTA
GTTTTTAAAG
AAATTGTATT
TGTTAAATAT
GTACTACAAA
CTTAGTAGT