名称: | pBE2R-mRFP1 |
---|---|
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: | |
原核抗性: | Amp |
筛选标记: | |
荧光标记: | 红色 |
启动子: | p43 |
复制子: | repB |
拷贝数: | 低 |
感受态: | DH5a |
温度: | 37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。
GATCTCAACA
GCGGTAAGAT
CCTTGAGAGT
TTTCGCCCCG
AAGAACGTTT
TCCAATGATG
AGCACTTTTA
AAGTTCTGCT
ATGTGGCGCG
GTATTATCCC
GTATTGACGC
CGGGCAAGAG
CAACTCGGTC
GCCGCATACA
CTATTCTCAG
AATGACTTGG
TTGAGTACTC
ACCAGTCACA
GAAAAGCATC
TTACGGATGG
CATGACAGTA
AGAGAATTAT
GCAGTGCTGC
CATAACCATG
AGTGATAACA
CTGCGGCCAA
CTTACTTCTG
ACAACGATCG
GAGGACCGAA
GGAGCTAACC
GCTTTTTTGC
ACAACATGGG
GGATCATGTA
ACTCGCCTTG
ATCGTTGGGA
ACCGGAGCTG
AATGAAGCCA
TACCAAACGA
CGAGCGTGAC
ACCACGATGC
CTGTAGCAAT
GGCAACAACG
TTGCGCAAAC
TATTAACTGG
CGAACTACTT
ACTCTAGCTT
CCCGGCAACA
ATTAATAGAC
TGGATGGAGG
CGGATAAAGT
TGCAGGACCA
CTTCTGCGCT
CGGCCCTTCC
GGCTGGCTGG
TTTATTGCTG
ATAAATCTGG
AGCCGGTGAG
CGTGGGTCTC
GCGGTATCAT
TGCAGCACTG
GGGCCAGATG
GTAAGCCCTC
CCGTATCGTA
GTTATCTACA
CGACGGGGAG
TCAGGCAACT
ATGGATGAAC
GAAATAGACA
GATCGCTGAG
ATAGGTGCCT
CACTGATTAA
GCATTGGTAA
CTGTCAGACC
AAGTTTACTC
ATATATACTT
TAGATTGATT
TAAAACTTCA
TTTTTAATTT
AAAAGGATCT
AGGTGAAGAT
CCTTTTTGAT
AATCTCATGA
CCAAAATCCC
TTAACGTGAG
TTTTCGTTCC
ACTGAGCGTC
AGACCCCGTA
GAAAAGATCA
AAGGATCTTC
TTGAGATCCT
TTTTTTCTGC
GCGTAATCTG
CTGCTTGCAA
ACAAAAAAAC
CACCGCTACC
AGCGGTGGTT
TGTTTGCCGG
ATCAAGAGCT
ACCAACTCTT
TTTCCGAAGG
TAACTGGCTT
CAGCAGAGCG
CAGATACCAA
ATACTGTCCT
TCTAGTGTAG
CCGTAGTTAG
GCCACCACTT
CAAGAACTCT
GTAGCACCGC
CTACATACCT
CGCTCTGCTA
ATCCTGTTAC
CAGTGGCTGC
TGCCAGTGGC
GATAAGTCGT
GTCTTACCGG
GTTGGACTCA
AGACGATAGT
TACCGGATAA
GGCGCAGCGG
TCGGGCTGAA
CGGGGGGTTC
GTGCACACAG
CCCAGCTTGG
AGCGAACGAC
CTACACCGAA
CTGAGATACC
TACAGCGTGA
GCTATGAGAA
AGCGCCACGC
TTCCCGAAGG
GAGAAAGGCG
GACAGGTATC
CGGTAAGCGG
CAGGGTCGGA
ACAGGAGAGC
GCACGAGGGA
GCTTCCAGGG
GGAAACGCCT
GGTATCTTTA
TAGTCCTGTC
GGGTTTCGCC
ACCTCTGACT
TGAGCGTCGA
TTTTTGTGAT
GCTCGTCAGG
GGGGCGGAGC
CTATGGAAAA
ACGCCAGCAA
CGCGGCCTTT
TTACGGTTCC
TGGCCTTTTG
CTGGCCTTTT
GCTCACATGT
TCTTTCCTGC
GTTATCCCCT
GATTCTGTGG
ATAACCGTAT
TACCGCCTTT
GAGTGAGCTG
ATACCGCTCG
CCGCAGCCGA
ACGACCGAGC
GCAGCGAGTC
AGTGAGCGAG
GAAGCGGAAG
AGCGCCCAAT
ACGCAAACCG
CCTCTCCCCG
CGCGTTGGCC
GATTCATTAA
TGCAGCTGAT
TTCACTTTTT
GCATTCTACA
AACTGCATAA
CTCATATGTA
AATCGCTCCT
TTTTAGGTGG
CACAAATGTG
AGGCATTTTC
GCTCTTTCCG
GCAACCACTT
CCAAGTAAAG
TATAACACAC
TATACTTTAT
ATTCATAAAG
TGTGTGCTCT
GCGAGGCTGT
CGGCAGTGCC
GACCAAAACC
ATAAAACCTT
TAAGACCTTT
CTTTTTTTTA
CGAGAAAAAA
GAAACAAAAA
AACCTGCCCT
CTGCCACCTC
AGCAAAGGGG
GGTTTTGCTC
TCGTGCTCGT
TTAAAAATCA
GCAAGGGACA
GGTAGTATTT
TTTGAGAAGA
TCACTCAAAA
AATCTCCACC
TTTAAACCCT
TGCCAATTTT
TATTTTGTCC
GTTTTGTCTA
GCTTACCGAA
AGCCAGACTC
AGCAAGAATA
AAATTTTTAT
TGTCTTTCGG
TTTTCTAGTG
TAACGGACAA
AACCACTCAA
AATAAAAAAG
ATACAAGAGA
GGTCTCTCGT
ATCTTTTATT
CAGCAATCGC
GCCCGATTGC
TGAACAGATT
AATAATAGAT
TTTAGCTTTT
TATTTGTTGA
AAAAAGCTAA
TCAAATTGTT
GTCGGGATCA
ATTACTGCAA
AGTCTCGTTC
ATCCCACCAC
TGATCTTTTA
ATGATGTATT
GGGGTGCAAA
ATGCCCAAAG
GCTTAATATG
TTGATATAAT
TCATCAATTC
CCTCTACTTC
AATGCGGCAA
CTAGCAGTAC
CAGCAATAAA
CGACTCCGCA
CCTGTACAAA
CCGGTGAATC
ATTACTACGA
GAGCGCCAGC
TTCATCACTT
GCCTCCCATA
GATGAATCCG
AACCTCATTA
CACATTAGAA
CTGCGAATCC
ATCTTCATGG
TGAACCAAAG
TGAAACCTAG
TTTATCGCAA
TAAAAACCTA
TACTCTTTTT
AATATCCCCG
ACTGGCAATG
CCGGGATAGA
CTGTAACATT
CTCACGCATA
AAATCCCCTT
TCATTTTCTA
ATGTAAATCT
ATTACCTTAT
TATTAATTCA
ATTCGCTCAT
AATTAATCCT
TTTTCTTATT
ACGCAAAATG
GCCCGATTTA
AGCACACCCT
TTATTCCGTT
AATGCGCCAT
GACAGCCATG
ATAATTACTA
ATACTAGGAG
AAGTTAATAA
ATACGTAACC
AACATGATTA
ACAATTATTA
GAGGTCATCG
TTCAAAATGG
TATGCGTTTT
GACACATCCA
CTATATATCC
GTGTCGTTCT
GTCCACTCCT
GAATCCCATT
CCAGAAATTC
TCTAGCGATT
CCAGAAGTTT
CTCAGAGTCG
GAAAGTTGAC
CAGACATTAC
GAACTGGCAC
AGATGGTCAT
AACCTGAAGG
AAGATCTGAT
TGCTTAACTG
CTTCAGTTAA
GACCGAAGCG
CTCGTCGTAT
AACAGATGCG
ATGATGCAGA
CCAATCAACA
TGGCACCTGC
CATTGCTACC
TGTACAGTCA
AGGATGGTAG
AAATGTTGTC
GGTCCTTGCA
CACGAATATT
ACGCCATTTG
CCTGCATATT
CAAACAGCTC
TTCTACGATA
AGGGCACAAA
TCGCATCGTG
GAACGTTTGG
GCTTCTACCG
ATTTAGCAGT
TTGATACACT
TTCTCTAAGT
ATCCACCTGA
ATCATAAATC
GGCAAAATAG
AGAAAAATTG
ACCATGTGTA
AGCGGCCAAT
CTGATTCCAC
CTGAGATGCA
TAATCTAGTA
GAATCTCTTC
GCTATCAAAA
TTCACTTCCA
CCTTCCACTC
ACCGGTTGTC
CATTCATGGC
TGAACTCTGC
TTCCTCTGTT
GACATGACAC
ACATCATCTC
AATATCCGAA
TAGGGCCACA
GTCGGCATTA
TCTCATATTA
TAAAAGCCAG
TCATTAGGCC
TATCTGACAA
TTCCTGAATA
GAGTTCATAA
ACAATCCTGC
ATGATAACCA
TCACAAACAG
AATGATGTAC
CTGTAAAGAT
AGCGGTAAAT
ATATTGAATT
ACCTTTATTA
ATGAATTTTC
CTGCTGTAAT
AATGGGTAGA
AGGTAATTAC
TATTATTATT
GATATTTAAG
TTAAACCCAG
TAAATGAAGT
CCATGGAATA
ATAGAAAGAG
AAAAAGCATT
TTCAGGTATA
GGTGTTTTGG
GAAACAATTT
CCCCGAACCA
TTATATTTCT
CTACATCAGA
AAGGTATAAA
TCATAAAACT
CTTTGAAGTC
ATTCTTTACA
GGAGTCCAAA
TACCAGAGAA
TGTTTTAGAT
ACACCATCAA
AAATTGTATA
AAGTGGCTCT
AACTTATCCC
AATAACCTAA
CTCTCCGTCG
CTATTGTAAC
CAGTTCTAAA
AGCTGTATTT
GAGTTTATCA
CCCTTGTCAC
TAAGAAAATA
AATGCAGGGT
AAAATTTATA
TCCTTCTTGT
TTTATGTTTC
GGTATAAAAC
ACTAATATCA
ATTTCTGTGG
TTATACTAAA
AGTCGTTTGT
TGGTTCAAAT
AATGATTAAA
TATCTCTTTT
CTCTTCCAAT
TGTCTAAATC
AATTTTATTA
AAGTTCATTT
GATATGCCTC
CTGGCCCATC
AGTCTGACGA
CCAAGAGAGC
CATAAACACC
AATAGCCTTA
ACATCATCCC
CATATTTATC
CAATATTCGT
TCCTTAATTT
CATGAACAAT
CTTCATTCTT
TCTTCTCTAG
TCATTATTAT
TGGTCCATTC
ACTATTCTCA
TTCCCTTTTC
AGATAATTTT
AGATTTGCTT
TTCTAAATAA
GAATATTTGG
AGAGCACCGT
TCTTATTCAG
CTATTAATAA
CTCGTCTTCC
TAAGCATCCT
TCAATCCTTT
TAATAACAAT
TATAGCATCT
AATCTTCAAC
AAACTGGCCC
GTTTGTTGAA
CTACTCTTTA
ATAAAATAAT
TTTTCCGTTC
CCAATTCCAC
ATTGCAATAA
TAGAAAATCC
ATCTTCATCG
GCTTTTTCGT
CATCATCTGT
ATGAATCAAA
TCGCCTTCTT
CTGTGTCATC
AAGGTTTAAT
TTTTTATGTA
TTTCTTTTAA
CAAACCACCA
TAGGAGATTA
ACCTTTTACG
GTGTAAACCT
TCCTCCAAAT
CAGACAAACG
TTTCAAATTC
TTTTCTTCAT
CATCGGTCAT
AAAATCCGTA
TCCTTTACAG
GATATTTTGC
AGTTTCGTCA
ATTGCCGATT
GTATATCCGA
TTTATATTTA
TTTTTCGGTC
GAATCATTTG
AACTTTTACA
TTTGGATCAT
AGTCTAATTT
CATTGCCTTT
TTCCAAAATT
GAATCCATTG
TTTTTGATTC
ACGTAGTTTT
CTGTATTCTT
AAAATAAGTT
GGTTCCACAC
ATACCAATAC
ATGCATGTGC
TGATTATAAG
AATTATCTTT
ATTATTTATT
GTCACTTCCG
TTGCACGCAT
AAAACCAACA
AGATTTTTAT
TAATTTTTTT
ATATTGCATC
ATTCGGCGAA
ATCCTTGAGC
CATATCTGAC
AAACTCTTAT
TTAATTCTTC
GCCATCATAA
ACATTTTTAA
CTGTTAATGT
GAGAAACAAC
CAACGAACTG
TTGGCTTTTG
TTTAATAACT
TCAGCAACAA
CCTTTTGTGA
CTGAATGCCA
TGTTTCATTG
CTCTCCTCCA
GTTGCACATT
GGACAAAGCC
TGGATTTACA
AAACCACACT
CGATACAACT
TTCTTTCGCC
TGTTTCACGA
TTTTGTTTAT
ACTCTAATAT
TTCAGCACAA
TCTTTTACTC
TTTCAGCCTT
TTTAAATTCA
AGAATATGCA
GAAGTTCAAA
GTAATCAACA
TTAGCGATTT
TCTTTTCTCT
CCATGGTCTC
ACTTTTCCAC
TTTTTGTCTT
GTCCACTAAA
ACCCTTGATT
TTTCATCTGA
ATAAATGCTA
CTATTAGGAC
ACATAATATT
AAAAGAAACC
CCCATCTATT
TAGTTATTTG
TTTAGTCACT
TATAACTTTA
ACAGATGGGG
TTTTTCTGTG
CAACCAATTT
TAAGGGTTTT
CAATACTTTA
AAACACATAC
ATACCAACAC
TTCAACGCAC
CTTTCAGCAA
CTAAAATAAA
AATGACGTTA
TTTCTATATG
TATCAAGATA
AGAAAGAACA
AGTTCAAAAC
CATCAAAAAA
AGACACCTTT
TCAGGTGCTT
TTTTTATTTT
ATAAACTCAT
TCCCTGATCT
CGACTTCGTT
CTTTTTTTAC
CTCTCGGTTA
TGAGTTAGTT
CAAATTCGTT
CTTTTTAGGT
TCTAAATCGT
GTTTTTCTTG
GAATTGTGCT
GTTTTATCCT
TTACCTTGTC
TACAAACCCC
TTAAAAACGT
TTTTAAAGGC
TTTTAAGCCG
TCTGTACGTT
CCTTAAGGAA
TTCGAGCTCA
GCATTATTGA
GTGGATGATT
ATATTCCTTT
TGATAGGTGG
TATGTTTTCG
CTTGAACTTT
TAAATACAGC
CATTGAACAT
ACGGTTGATT
TAATAACTGA
CAAACATCAC
CCTCTTGCTA
AAGCGGCCAA
GGACGCTGCC
GCCGGGGCTG
TTTGCGTTTT
TGCCGTGATT
TCGTGTATTA
TTGGTTTACT
TATTTTTTTG
CCAAAGCTGT
AATGGCTGAA
AATTCTTACA
TTTATTTTAC
ATTTTTAGAA
ATGGGCGTGA
AAAAAAGCGC
GCGATTATGT
AAAATATAAA
GTGATAGCGG
TACCATTATA
CACACATACT
AGAGAAAGAG
GAGAAATACT
AGATGGCTTC
CTCCGAAGAC
GTTATCAAAG
AGTTCATGCG
TTTCAAAGTT
CGTATGGAAG
GTTCCGTTAA
CGGTCACGAG
TTCGAAATCG
AAGGTGAAGG
TGAAGGTCGT
CCGTACGAAG
GTACCCAGAC
CGCTAAACTG
AAAGTTACCA
AAGGTGGTCC
GCTGCCGTTC
GCTTGGGACA
TCCTGTCCCC
GCAGTTCCAG
TACGGTTCCA
AAGCTTACGT
TAAACACCCG
GCTGACATCC
CGGACTACCT
GAAACTGTCC
TTCCCGGAAG
GTTTCAAATG
GGAACGTGTT
ATGAACTTCG
AAGACGGTGG
TGTTGTTACC
GTTACCCAGG
ACTCCTCCCT
GCAAGACGGT
GAGTTCATCT
ACAAAGTTAA
ACTGCGTGGT
ACCAACTTCC
CGTCCGACGG
TCCGGTTATG
CAGAAAAAAA
CCATGGGTTG
GGAAGCTTCC
ACCGAACGTA
TGTACCCGGA
AGACGGTGCT
CTGAAAGGTG
AAATCAAAAT
GCGTCTGAAA
CTGAAAGACG
GTGGTCACTA
CGACGCTGAA
GTTAAAACCA
CCTACATGGC
TAAAAAACCG
GTTCAGCTGC
CGGGTGCTTA
CAAAACCGAC
ATCAAACTGG
ACATCACCTC
CCACAACGAA
GACTACACCA
TCGTTGAACA
GTACGAACGT
GCTGAAGGTC
GTCACTCCAC
CGGTGCTTAA
TAACGCTGAT
AGTGCTAGTG
TAGATCGCTA
CTAGAGCCAG
GCATCAAATA
AAACGAAAGG
CTCAGTCGAA
AGACTGGGCC
TTTCGTTTTA
TCTGTTGTTT
GTCGGTGAAC
GCTCTCTACT
AGAGTCACAC
TGGCTCACCT
TCGGGTGGGC
CTTTCTGCGT
TTATATAGGC
CATCTGGGCC
GGGGGCGTCT
CTGAGCTAGA
AATAGCAAGC
TTGTATTCTA
TAGTGTCACC
TAAATCGTAT
GTGTATGATA
CATAAGGTTA
TGTATTAATT
GTAGCCGCGT
TCTAACGACA
ATATGTACAA
GCCTAATTGT
GTAGCATCTG
GCTTACTGAA
GCAGACCCTA
TCATCTCTCT
CGTAAACTGC
CGTCAGAGTC
GGTTTGGTTG
GACGAACCTT
CTGAGTTTCT
GGTAACGCCG
TCCCGCACCC
GGAAATGGTC
AGCGAACCAA
TCAGCAGGGT
CATCGCTAGC
CAGATCCTCT
ACGCCGGACG
CATCGTGGCC
GGCATCACCG
GCGCCACAGG
TGCGGTTGCT
GGCGCCTATA
TCGCCGACAT
CACCGATGGG
GAAGATCGGG
CTCGCCACTT
CGGGCTCATG
AGCGCTTGTT
TCGGCGTGGG
TATGGTGGCA
GGCCCCGTGG
CCGGGGGACT
GTTGGGCGCC
ATCTCCTTGC
ACCATTCCTT
GCGGCGGCGG
TGCTCAACGG
CCTCAACCTA
CTACTGGGCT
GCTTCCTAAT
GCAGGAGTCG
CATAAGGGAG
AGCGTCGATA
TGGTGCACTC
TCAGTACAAT
CTGCTCTGAT
GCCGCATAGT
TAAGCCAGCC
CCGACACCCG
CCAACACCCG
CTGACGCGCC
CTGACGGGCT
TGTCTGCTCC
CGGCATCCGC
TTACAGACAA
GCTGTGACCG
TCTCCGGGAG
CTGCATGTGT
CAGAGGTTTT
CACCGTCATC
ACCGAAACGC
GCGAGACGAA
AGGGCCTCGT
GATACGCCTA
TTTTTATAGG
TTAATGTCAT
GATAATAATG
GTTTCTTAGA
CGTCAGGTGG
CACTTTTCGG
GGAAATGTGC
GCGGAACCCC
TATTTGTTTA
TTTTTCTAAA
TACATTCAAA
TATGTATCCG
CTCATGAGAC
AATAACCCTG
ATAAATGCTT
CAATAATATT
GAAAAAGGAA
GAGTATGAGT
ATTCAACATT
TCCGTGTCGC
CCTTATTCCC
TTTTTTGCGG
CATTTTGCCT
TCCTGTTTTT
GCTCACCCAG
AAACGCTGGT
GAAAGTAAAA
GATGCTGAAG
ATCAGTTGGG
TGCACGAGTG
GGTTACATCG
AACTG