名称: | pBE2R-mCitrine |
---|---|
别称: | |
基因长: | |
质粒宿主: | 枯草杆菌 |
质粒用途: | 蛋白表达,分泌表达 |
片段类型: | ORF |
片段物种: | 空载体 |
原核抗性: | Amp |
筛选标记: | Kan |
荧光标记: | 黄色 |
启动子: | p43 |
复制子: | repB |
拷贝数: | 低 |
感受态: | Stbl3 |
温度: | 30度 |
背景载体: | |
正向引物: | |
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。
AGATTAATAA
TAGATTTTAG
CTTTTTATTT
GTTGAAAAAA
GCTAATCAAA
TTGTTGTCGG
GATCAATTAC
TGCAAAGTCT
CGTTCATCCC
ACCACTGATC
TTTTAATGAT
GTATTGGGGT
GCAAAATGCC
CAAAGGCTTA
ATATGTTGAT
ATAATTCATC
AATTCCCTCT
ACTTCAATGC
GGCAACTAGC
AGTACCAGCA
ATAAACGACT
CCGCACCTGT
ACAAACCGGT
GAATCATTAC
TACGAGAGCG
CCAGCTTCAT
CACTTGCCTC
CCATAGATGA
ATCCGAACCT
CATTACACAT
TAGAACTGCG
AATCCATCTT
CATGGTGAAC
CAAAGTGAAA
CCTAGTTTAT
CGCAATAAAA
ACCTATACTC
TTTTTAATAT
CCCCGACTGG
CAATGCCGGG
ATAGACTGTA
ACATTCTCAC
GCATAAAATC
CCCTTTCATT
TTCTAATGTA
AATCTATTAC
CTTATTATTA
ATTCAATTCG
CTCATAATTA
ATCCTTTTTC
TTATTACGCA
AAATGGCCCG
ATTTAAGCAC
ACCCTTTATT
CCGTTAATGC
GCCATGACAG
CCATGATAAT
TACTAATACT
AGGAGAAGTT
AATAAATACG
TAACCAACAT
GATTAACAAT
TATTAGAGGT
CATCGTTCAA
AATGGTATGC
GTTTTGACAC
ATCCACTATA
TATCCGTGTC
GTTCTGTCCA
CTCCTGAATC
CCATTCCAGA
AATTCTCTAG
CGATTCCAGA
AGTTTCTCAG
AGTCGGAAAG
TTGACCAGAC
ATTACGAACT
GGCACAGATG
GTCATAACCT
GAAGGAAGAT
CTGATTGCTT
AACTGCTTCA
GTTAAGACCG
AAGCGCTCGT
CGTATAACAG
ATGCGATGAT
GCAGACCAAT
CAACATGGCA
CCTGCCATTG
CTACCTGTAC
AGTCAAGGAT
GGTAGAAATG
TTGTCGGTCC
TTGCACACGA
ATATTACGCC
ATTTGCCTGC
ATATTCAAAC
AGCTCTTCTA
CGATAAGGGC
ACAAATCGCA
TCGTGGAACG
TTTGGGCTTC
TACCGATTTA
GCAGTTTGAT
ACACTTTCTC
TAAGTATCCA
CCTGAATCAT
AAATCGGCAA
AATAGAGAAA
AATTGACCAT
GTGTAAGCGG
CCAATCTGAT
TCCACCTGAG
ATGCATAATC
TAGTAGAATC
TCTTCGCTAT
CAAAATTCAC
TTCCACCTTC
CACTCACCGG
TTGTCCATTC
ATGGCTGAAC
TCTGCTTCCT
CTGTTGACAT
GACACACATC
ATCTCAATAT
CCGAATAGGG
CCCATCAGTC
TGACGACCAA
GAGAGCCATA
AACACCAATA
GCCTTAACAT
CATCCCCATA
TTTATCCAAT
ATTCGTTCCT
TAATTTCATG
AACAATCTTC
ATTCTTTCTT
CTCTAGTCAT
TATTATTGGT
CCATTCACTA
TTCTCATTCC
CTTTTCAGAT
AATTTTAGAT
TTGCTTTTCT
AAATAAGAAT
ATTTGGAGAG
CACCGTTCTT
ATTCAGCTAT
TAATAACTCG
TCTTCCTAAG
CATCCTTCAA
TCCTTTTAAT
AACAATTATA
GCATCTAATC
TTCAACAAAC
TGGCCCGTTT
GTTGAACTAC
TCTTTAATAA
AATAATTTTT
CCGTTCCCAA
TTCCACATTG
CAATAATAGA
AAATCCATCT
TCATCGGCTT
TTTCGTCATC
ATCTGTATGA
ATCAAATCGC
CTTCTTCTGT
GTCATCAAGG
TTTAATTTTT
TATGTATTTC
TTTTAACAAA
CCACCATAGG
AGATTAACCT
TTTACGGTGT
AAACCTTCCT
CCAAATCAGA
CAAACGTTTC
AAATTCTTTT
CTTCATCATC
GGTCATAAAA
TCCGTATCCT
TTACAGGATA
TTTTGCAGTT
TCGTCAATTG
CCGATTGTAT
ATCCGATTTA
TATTTATTTT
TCGGTCGAAT
CATTTGAACT
TTTACATTTG
GATCATAGTC
TAATTTCATT
GCCTTTTTCC
AAAATTGAAT
CCATTGTTTT
TGATTCACGT
AGTTTTCTGT
ATTCTTAAAA
TAAGTTGGTT
CCACACATAC
CAATACATGC
ATGTGCTGAT
TATAAGAATT
ATCTTTATTA
TTTATTGTCA
CTTCCGTTGC
ACGCATAAAA
CCAACAAGAT
TTTTATTAAT
TTTTTTATAT
TGCATCATTC
GGCGAAATCC
TTGAGCCATA
TCTGACAAAC
TCTTATTTAA
TTCTTCGCCA
TCATAAACAT
TTTTAACTGT
TAATGTGAGA
AACAACCAAC
GAACTGTTGG
CTTTTGTTTA
ATAACTTCAG
CAACAACCTT
TTGTGACTGA
ATGCCATGTT
TCATTGCTCT
CCTCCAGTTG
CACATTGGAC
AAAGCCTGGA
TTTACAAAAC
CACACTCGAT
ACAACTTTCT
TTCGCCTGTT
TCACGATTTT
GTTTATACTC
TAATATTTCA
GCACAATCTT
TTACTCTTTC
AGCCTTTTTA
AATTCAAGAA
TATGCAGAAG
TTCAAAGTAA
TCAACATTAG
CGATTTTCTT
TTCTCTCCAT
GGTCTCACTT
TTCCACTTTT
TGTCTTGTCC
ACTAAAACCC
TTGATTTTTC
ATCTGAATAA
ATGCTACTAT
TAGGACACAT
AATATTAAAA
GAAACCCCCA
TCTATTTAGT
TATTTGTTTA
GTCACTTATA
ACTTTAACAG
ATGGGGTTTT
TCTGTGCAAC
CAATTTTAAG
GGTTTTCAAT
ACTTTAAAAC
ACATACATAC
CAACACTTCA
ACGCACCTTT
CAGCAACTAA
AATAAAAATG
ACGTTATTTC
TATATGTATC
AAGATAAGAA
AGAACAAGTT
CAAAACCATC
AAAAAAAGAC
ACCTTTTCAG
GTGCTTTTTT
TATTTTATAA
ACTCATTCCC
TGATCTCGAC
TTCGTTCTTT
TTTTACCTCT
CGGTTATGAG
TTAGTTCAAA
TTCGTTCTTT
TTAGGTTCTA
AATCGTGTTT
TTCTTGGAAT
TGTGCTGTTT
TATCCTTTAC
CTTGTCTACA
AACCCCTTAA
AAACGTTTTT
AAAGGCTTTT
AAGCCGTCTG
TACGTTCCTT
AAGGAATTCG
AGCTCAGCAT
TATTGAGTGG
ATGATTATAT
TCCTTTTGAT
AGGTGGTATG
TTTTCGCTTG
AACTTTTAAA
TACAGCCATT
GAACATACGG
TTGATTTAAT
AACTGACAAA
CATCACCCTC
TTGCTAAAGC
GGCCAAGGAC
GCTGCCGCCG
GGGCTGTTTG
CGTTTTTGCC
GTGATTTCGT
GTATTATTGG
TTTACTTATT
TTTTTGCCAA
AGCTGTAATG
GCTGAAAATT
CTTACATTTA
TTTTACATTT
TTAGAAATGG
GCGTGAAAAA
AAGCGCGCGA
TTATGTAAAA
TATAAAGTGA
TAGCGGTACA
AGAGAGGAAT
GTACACATGA
GCAAAGGCGA
AGAACTGTTT
ACCGGTGTTG
TTCCGATTCT
GGTTGAACTG
GATGGTGATG
TTAATGGCCA
CAAATTTTCA
GTTAGCGGTG
AAGGCGAAGG
TGATGCAACC
TATGGTAAAC
TGACCCTGAA
ATTTATCTGT
ACCACCGGCA
AACTGCCGGT
TCCGTGGCCG
ACACTGGTTA
CCACCTTTGG
TTATGGTCTG
ATGTGTTTTG
CACGTTATCC
GGATCATATG
AAACAGCACG
ATTTTTTCAA
AAGCGCAATG
CCGGAAGGTT
ATGTTCAAGA
ACGTACCATC
TTCTTCAAAG
ATGACGGCAA
CTATAAAACC
CGTGCCGAAG
TTAAATTTGA
AGGTGATACC
CTGGTGAATC
GCATTGAACT
GAAAGGCATC
GATTTTAAAG
AGGATGGTAA
TATCCTGGGC
CACAAACTGG
AATATAATTA
TAATAGCCAC
AACGTGTACA
TCATGGCCGA
CAAACAGAAA
AATGGCATCA
AAGTGAACTT
CAAGATCCGC
CATAATATTG
AAGATGGTTC
AGTTCAGCTG
GCCGATCATT
ATCAGCAGAA
TACCCCGATT
GGTGATGGTC
CGGTTCTGCT
GCCGGATAAT
CATTATCTGA
GCTATCAGAG
CAAACTGAGC
AAAGATCCGA
ATGAAAAACG
TGATCACATG
GTGCTGCTGG
AATTTGTTAC
CGCAGCAGGT
ATTACCTTAG
GTATGGATGA
ACTGTACAAA
TAATGTATTC
TATAGTGTCA
CCTAAATCGT
ATGTGTATGA
TACATAAGGT
TATGTATTAA
TTGTAGCCGC
GTTCTAACGA
CAATATGTAC
AAGCCTAATT
GTGTAGCATC
TGGCTTACTG
AAGCAGACCC
TATCATCTCT
CTCGTAAACT
GCCGTCAGAG
TCGGTTTGGT
TGGACGAACC
TTCTGAGTTT
CTGGTAACGC
CGTCCCGCAC
CCGGAAATGG
TCAGCGAACC
AATCAGCAGG
GTCATCGCTA
GCCAGATCCT
CTACGCCGGA
CGCATCGTGG
CCGGCATCAC
CGGCGCCACA
GGTGCGGTTG
CTGGCGCCTA
TATCGCCGAC
ATCACCGATG
GGGAAGATCG
GGCTCGCCAC
TTCGGGCTCA
TGAGCGCTTG
TTTCGGCGTG
GGTATGGTGG
CAGGCCCCGT
GGCCGGGGGA
CTGTTGGGCG
CCATCTCCTT
GCACCATTCC
TTGCGGCGGC
GGTGCTCAAC
GGCCTCAACC
TACTACTGGG
CTGCTTCCTA
ATGCAGGAGT
CGCATAAGGG
AGAGCGTCGA
TATGGTGCAC
TCTCAGTACA
ATCTGCTCTG
ATGCCGCATA
GTTAAGCCAG
CCCCGACACC
CGCCAACACC
CGCTGACGCG
CCCTGACGGG
CTTGTCTGCT
CCCGGCATCC
GCTTACAGAC
AAGCTGTGAC
CGTCTCCGGG
AGCTGCATGT
GTCAGAGGTT
TTCACCGTCA
TCACCGAAAC
GCGCGAGACG
AAAGGGCCTC
GTGATACGCC
TATTTTTATA
GGTTAATGTC
ATGATAATAA
TGGTTTCTTA
GACGTCAGGT
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
CTGTCCTTCT
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
AGCTGATTTC
ACTTTTTGCA
TTCTACAAAC
TGCATAACTC
ATATGTAAAT
CGCTCCTTTT
TAGGTGGCAC
AAATGTGAGG
CATTTTCGCT
CTTTCCGGCA
ACCACTTCCA
AGTAAAGTAT
AACACACTAT
ACTTTATATT
CATAAAGTGT
GTGCTCTGCG
AGGCTGTCGG
CAGTGCCGAC
CAAAACCATA
AAACCTTTAA
GACCTTTCTT
TTTTTTACGA
GAAAAAAGAA
ACAAAAAAAC
CTGCCCTCTG
CCACCTCAGC
AAAGGGGGGT
TTTGCTCTCG
TGCTCGTTTA
AAAATCAGCA
AGGGACAGGT
AGTATTTTTT
GAGAAGATCA
CTCAAAAAAT
CTCCACCTTT
AAACCCTTGC
CAATTTTTAT
TTTGTCCGTT
TTGTCTAGCT
TACCGAAAGC
CAGACTCAGC
AAGAATAAAA
TTTTTATTGT
CTTTCGGTTT
TCTAGTGTAA
CGGACAAAAC
CACTCAAAAT
AAAAAAGATA
CAAGAGAGGT
CTCTCGTATC
TTTTATTCAG
CAATCGCGCC
CGATTGCTGA
AC