名称: | pDL278-P23-FLAG-htrA |
---|---|
别称: | |
基因长: | 1227bp |
质粒宿主: | 革兰氏阳性菌;粪肠球菌 |
质粒用途: | 蛋白表达 |
片段类型: | ORF |
片段物种: | |
原核抗性: | Spe |
筛选标记: | |
荧光标记: | |
启动子: | P23 |
复制子: | pUC |
拷贝数: | |
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。
AAATACGTTT
CCCATTTAGC
CGTCATTTCA
GCACTCGTTA
AAAGGTGCTG
ACTTTCAACT
GCCTGGCACA
ATAATTTTCC
TTTTTCAGTT
ACAACAAGCT
CCAATACGCA
AACCGCCTCT
CCCCGCGCGT
TGGCCGATTC
ATTAATGCAG
CTGGCACGAC
AGGTTTCCCG
ACTGGAAAGC
GGGCAGTGAG
CGCAACGCAA
TTAATGTGAG
TTAGCTCACT
CATTAGGCAC
CCCAGGCTTT
ACACTTTATG
CTTCCGGCTC
GTATGTTGTG
TGGAATTGTG
AGCGGATAAC
AATTTCACAC
AGGAAACAGC
TATGACCATG
ATTACGCCAA
GCTTGAAAAG
CCCTGACAAC
CCTTGTTCCT
AAAAAGGAAT
AAGCGTTCGG
TCAGTAAATA
ATAGAAATAA
AAAATCAGAC
CTAAGACTGA
TGACAAAAAG
AGAAAATTTT
GATAAAATAG
TCTTAGAATT
AAATTAAAAA
GGGAGGCCAA
ATATAATGAA
AAATATGAAT
GACAATGATG
TTCATATGGA
CTACAAGGAC
GACGATGACA
AGGGAGGTGG
CGGTTCACTC
GAGGTGAAAA
AAAAATTAGT
CTCATCACTT
CTAAAGTGTT
CTCTAATCAT
TATTGTTAGC
TTTGCTGGTG
GAGCATTTGC
TAGTTTTGTC
ATGAATCATA
ATGACAATAT
TCCAAATGGT
GGTGTCACTA
AAACTAGTAA
AGTAAATTAT
AATAACATAA
CGCCTACAAC
AAAAGCTGTT
AAAAAGGTAC
AAAATAGTGT
TGTTTCTGTT
ATCAATTATA
AACAACAAGA
GAGTCGTTCT
GACCTATCAG
ACTTCTATAG
TCATTTTTTT
GGTAATCAGG
GGGGCAACAC
TGATAAGGGC
TTACAAGTTT
ACGGTGAAGG
CTCTGGAGTC
ATCTATAAAA
AAGATGGTAA
AAATGCCTAT
GTTGTCACTA
ATAACCACGT
CATTGATGGG
GCTAAACAAA
TTGAAATTCA
ACTAGCTGAT
GGCTCAAAAG
CAGTTGGGAA
ACTTGTTGGG
TCAGATACCT
ACTCTGATTT
AGCCGTCGTC
AAAATTCCAT
CAGATAAGGT
TTCAAATATT
GCAGAATTTG
CTGATTCATC
AAAACTCAAC
ATTGGTGAAA
CTGCTATAGC
GATCGGAAGC
CCTCTTGGAA
CTGAGTATGC
AAATTCTGTA
ACTCAAGGTA
TTGTATCTAG
TTTAAAAAGA
ACTGTAACAA
TGACTAATGA
AGAAGGACAA
ACAGTTTCTA
CAAATGCTAT
CCAGACTGAT
GCTGCTATCA
ATCCTGGTAA
TTCAGGTGGA
GCACTTATCA
ATATTGAAGG
ACAGGTTATT
GGAATTAATT
CTAGTAAAAT
TTCTTCTACA
TCAAATCAAA
CCTCAGGACA
ATCGTCAGGA
AATAGCGTTG
AAGGTATGGG
ATTTGCCATT
CCTTCAAATG
ATGTTGTTAA
GATTATCAAT
CAACTTGAGA
GTAACGGACA
AGTAGAGAGA
CCTGCTCTAG
GTATTTCTAT
GGCTGGATTA
AGTAATTTAC
CATCCGATGT
TATTAGTAAA
CTGAAAATCC
CAAGTAATGT
TACTAATGGT
ATTGTAGTAG
CATCTATCCA
ATCTGGCATG
CCAGCTCAAG
GCAAACTAAA
GAAATACGAT
GTCATTACTA
AAGTTGACGA
TAAAGAAGTA
GCATCTCCAA
GTGATTTACA
AAGTTTACTC
TATGGCCACC
AGGTAGGGGA
TTCCATAACA
GTAACCTTTT
ATCGTGGTGA
AAATAAACAA
ACAGTAACTA
TAAAACTTAC
TAAAACTAGT
AAAGATTTAG
CTAAACAACG
AGCAAATAAC
TAAGTCGACT
CTAGAGGATC
CCCGGGTACC
GAGCTCGAAT
TCACTGGCCG
TCGTTTTACA
ACGTCGTGAC
TGGGAAAACC
CTGGCGTTAC
CCAACTTAAT
CGCCTTGCAG
CACATCCCCC
TTTCGCCAGC
TGGCGTAATA
GCGAAGAGGC
CCGCACCGAT
CGCCCTTCCC
AACAGTTGCG
CAGCCTGAAT
GGCGAATGCC
GGGAATTATT
CACCACTCCA
AGAATTGGAG
CCAATCAATT
CTTGCGGAGA
ACTGTGAATG
CGCAAACCAA
CCCTTGGCAG
AACATATCCA
TCGCGTCCGC
CATCTCCAGC
AGCCGCACGC
GGCGCATCTC
GGCTGTTTTG
GCGGATGAGA
GAAGATTTTC
AGCCTGATAC
AGATTAAATC
AGAACGCAGA
AGCGGTCTGA
TAAAACAGAA
TTTGCCTGGC
GGCAGTAGCG
CGGTGGTCCC
ACCTGACCCC
ATGCCGAACT
CAGAAGTGAA
ACGCCGTAGC
GCCGATGGTA
GTGTGGGGTC
TCCCCATGCG
AGAGTAGGGA
ACTGCCAGGC
ATCAAATAAA
ACGAAAGGCT
CAGTCGAAAG
ACTGGGCCTT
TCGTTTTATC
TGTTGTTTGT
CGGTGAACGC
TCTCCTGAGT
AGGACAAATC
CGCCGGGAGC
GGATTTGAAC
GTTGCGAAGC
AACGGCCCGG
AGGGTGGCGG
GCAGGACGCC
CGCCATAAAC
TGCCAGGCAT
CAAATTAAGC
AGAAGGCCAT
CCTGACGGAT
GGCCTTTTTG
CGTTTCTACA
AACTCTTCCT
GTCGTCATAT
CTACAAGCCA
TCCCCCCACA
GATACGGTAA
ACTAGCCTCG
TTTTTGCATC
AGGAAAGCAG
CTGTTTTGGC
GGATGAGAGA
AGATTTTCAG
CCTGATACAG
ATTAAATCAG
AACGCAGAAG
CGGTCTGATA
AAACAGAATT
TGCCTGGCGG
CAGTAGCGCG
GTGGTCCCAC
CTGACCCCAT
GCCGAACTCA
GAAGTGAAAC
GCCGTAGCGC
CGATGGTAGT
GTGGGGTCTC
CCCATGCGAG
AGTAGGGAAC
TGCCAGGCAT
CAAATAAAAC
GAAAGGCTCA
GTCGAAAGAC
TGGGCCTTTC
GTTTTATCTG
TTGTTTGTCG
GTGAACGCTC
TCCTGAGTAG
GACAAATCCG
CCGGGAGCGG
ATTTGAACGT
TGCGAAGCAA
CGGCCCGGAG
GGTGGCGGGC
AGGACGCCCG
CCATAAACTG
CCAGGCATCA
AATTAAGCAG
AAGGCCATCC
TGACGGATGG
CCTTTTTGCG
TTTCTACAAA
CTCTTCCTGT
CGTCATATCT
ACAAGCCATC
CCCCCACAGA
TACGGTAAAC
TAGCCTCGTT
TTTGCATCAG
GAAAGCAGTC
GGGCAGCGTT
GGGTCCTGGC
GATTCTGAGA
CCTCTGAGAG
GCTCTCAGAG
CTATCTAAAG
CTGAGGGATA
TATAAATACC
TTAGAAAATT
ATTCGAAGAG
CTTAGAAGCG
AAAATAGAGC
GTTTAGAGCG
CGAGGGGCTG
AAATTAGAAA
AACTAAAAAC
ACAAATAGCT
GACCTAAAAA
TCATGTCTGA
GAAAGAACTA
GCGGCTATTA
CCCCTAAAAA
AGGCGTGTTC
GGTAAAGAAT
ATGTGGAATT
GACTAAAGAG
CAGTTTGAAG
AATTTAAAGG
GCTGATATAC
CGAAGCAGAA
ACCTTGTTCA
TCAAAAAGAG
CTAGAGAATG
AGCAATTAAG
GCGGATAGTG
CCTCTGAGAC
GCTCTAAACG
GTTTGAAGCG
AGTTGGAACG
AGCTAAAGAA
AAAAGTAAGG
GAGAGAGCAT
AGAGCGTCTT
AGGAACGAAA
ATAGAGCGCT
TAGAAGTGAA
AACTCAGTTT
TGAGACAGCA
AAATGACAAA
ATGCTAGGAA
AACTGAAAGA
GTTTATGCCA
GATAAAGCCT
TAAAAAATTT
TATATCAGAG
TTAAGAGCTA
TTCAGCCAAT
CGTAAGGGTA
GTTAAACGAG
TGATTGAAAA
AGGGCTAGGC
CTTTGAGCGA
TTTATGCCGT
GAAAGCTAAT
TGACAATAAG
CAAGGGCAAA
GTACGCTAGG
ACGTGACGAG
CCGAAAGGCT
TTAGCGTTTC
GAGCCGACAC
GGACAAAGGA
CGTCCGCCCT
TGGTTACTTG
TTGTCAATTA
GACCATGGAA
TAAAGTAAGC
GGACATGGTA
TAATAGCTAG
GTCGCAACGT
TCTTTCGCTA
AGTTACGAAC
TTAGATTGGA
GGTGAGCGCT
GTGAAGACTT
TCCTAGAACT
TGTTTTGATA
CCTTTTGTGG
TTGGCGTTGC
TGCAGAGGTA
AGTGCTGATT
GGTTGATTCG
GTATGTTCGA
AACAAACGCG
ACAAAAATTA
ACCTGAGTTC
TTTTTGAGGA
CAAAAAAGAA
AGACAGTAGT
TCCAGCTACT
GTTTTTTTTG
CGTTGTGCTA
TTCGTTTCCT
AGAACTTCTA
GCGTTAAAAT
TATTATACCA
CGTTTGGATT
TAGAAAGTCA
AAATTTGAGG
TTTTAGGGGT
GAATTTTTCG
TGAAACGAAA
AAGAGGGCTG
AAAAGCCCTT
AAAAACCCAA
TTGCGTAGCA
AGGGTTTTTT
CTTATCTTGA
TACATATAGA
AATAATGAGT
TTTTTTATTT
TCTTGTTTTA
AAGCACCTCA
AACCCTTGAT
ATTGCTGGGT
TTTTAGGTAA
CAAAAAAGCC
CTTGCAATTT
AATAAAATAA
AGTTTATAAT
TTAAGTGTCC
AATTTAAAAT
AATAAACTTA
GGAGAATTGC
AGGAACTTTT
TTATATACTC
AAAAAAATTT
TTTTGCAAGA
AAATTATAAC
ATGACAGGTA
CTGAAAATCA
AGTCTTTAAG
GACTATTCTA
AGAATGGCAA
AGATAGAAAG
TGGCGAGAAC
GCAAGTTAAA
AAATATTGAG
CTTGCTGGTC
GTTTAGAGCG
TTTAGGATAT
CGTTCGTTTG
AACGGGTCTA
TCAATGTGCC
GAAGTGTTGA
AGTTTATCGA
ACAACAAGAC
GGCACGAAAA
AACTCTATCA
GTCTTATTTT
TGCAAAAATA
AGCTCTGCCC
AATTTGTAAC
TGGAGACGGT
CAATGAAGTA
TGCTTATCAA
GCTGAATTAG
TGGTAAATGA
AGCAATGAAA
CGCTATCCAA
AAGGTCGCTT
TCTCTTTTTG
ACCTTGACGA
TTAAGAACAT
TAGTGGTGAA
AAATTAAATA
AATCAATTTC
AGAAATAGGG
CGAGCTTTTA
ATCGTCTTAT
GAAATACAAG
AAAGTCGATA
AAAATGTTAT
TGGCTATTTG
CGAGCCACTG
AGGTAACTTA
TTCAACTGAG
CATGAGAATT
ATCACCCTCA
TTTGCATGTA
TTGTTATTTG
TGAAATCTAG
CTATTTTACT
GGAAATAATA
CAAATTACAT
TAGCCAAGAA
GAATGGACGA
AACTATGGGC
TAAGGCGATG
AAATTGGATT
ATACACCTGT
AGTTGATATT
CGAACCGTCA
AAGCTCATAA
ACGTAAAAAC
TTGAAGTCAG
CTATTATCGA
AACAGCTAAA
TATCCTGTTA
AGCCTTTTGA
TGTAGATACA
GAAGATGTGA
CATTATTCTC
TGAAATGGTC
AAAGAACGGA
TAACAGAAGA
TTTAACGAAT
GGTTTGCACC
GAAAAAGGCA
GATTGGTTTT
GGAAAGTTGT
TCAAGAAAAT
CAAGGCGGAG
TTAGCTCTTG
ATGATGTCGA
AGAAGGGAAT
CTTGTTCAGA
CCGGAGCAGA
AGAATCTGCA
GAAAGTACTG
GTCGTGAAAT
TGTTGCCTTT
TGGAATTGGG
ATAGAAAGAA
TTATTTTGTG
AGGTAGCTGA
TGAGTAAAAC
AGTATCAGAA
TTAGCTCAAG
AATTGGGAGT
TAGTAGGCAG
TATCTTAATC
GGATTTTATC
GCAAAATAAT
CTCGGTCGAA
AAAAAGGGAA
TAAAAAAGTA
GTTTCCGATA
TGGACGAGAA
AGTTATGCAA
GGGTTTATTG
TTTTCTAAAA
TCTGATTACC
AATTAGAATG
AATATTTCCC
AAATATTAAA
TAATAAAACA
AAAAAATTGA
AAAAAGTGTT
TCCACCATTT
TTTCAATTTT
TTTATAATTT
TTTTAATCTG
TTATTTAAAT
AGTTTATAGT
TAAATTTACA
TTTTCATTAG
TCCATTCAAT
ATTCTCTCCA
AGATAACTAC
GAACTGCTAA
CAAAATTCTC
TCCCTATGTT
CTAATGGAGA
AGATTCAGCC
ACTGCATTTC
CCGCAATATC
TTTTGGTATG
ATTTTACCCG
TGTCCATAGT
TAAAATCATA
CGGCATAAAG
TTAATATAGA
GTTGGTTTCA
TCATCCTGAT
AATTATCTAT
TAATTCCTCT
GACGAATCCA
TAATGGCTCT
TCTCACATCA
GAAAATGGAA
TATCAGGTAG
TAATTCCTCT
AAGTCATAAT
TTCCGTATAT
TCTTTTATTT
TTTCGTTTTG
CTTGGTAAAG
CATTATGGTT
AAATCTGAAT
TTAATTCCTT
CTGAGGAATG
TATCCTTGTT
CATAAAGCTC
TTGTAACCAT
TCTCCATAAA
TAAATTCTTG
TTTGGGAGGA
TGATTCCACG
GTACCATTTC
TTGCTGAATA
ATAATTGTTA
ATTCAATATA
TCGTAAGTTG
CTTTTATCTC
CTATTTTTTT
TGAAATAGGT
CTAATTTTTT
GTATAAGTAT
TTCTTTACTT
TGATCTGTCA
ATGGTTCAGA
TACGACGACT
AAAAAGTCAA
GATCACTATT
TGGTTTTAGT
CCACTCTCAA
CTCCTGATCC
AAACATGTAA
GTACCAATAA
GGTTATTTTT
TAAATGTTTC
CGAAGTATTT
TTTTCACTTT
ATTAATTTGT
TCGTATGTAT
TCAAATATAT
CCTCCTCACT
ATTTTGATTA
GTACCTATTT
TATATCCATA
GTTGTTAATT
AAATAAACTT
AATTTAGTTT
ATTTATAGAT
TTCATTGGCT
TCTAAATTTT
TTATCTAGAT
AATAATTATT
TTAGTTAATT
TTATTCTAGA
TTATATATGA
TATGATCTTT
CATTTCCATA
AAACTAAAGT
AAGTGTAAAC
CTATTCATTG
TTTTAAAAAT
ATCTCTTGCC
AGTCACGTTA
CGTTATTAGT
TATAGTTATT
ATAACATGTA
TTCACGAACG
AAAATCGATC
TTCTATATAA
AAGATATATT
ATCTTATCAG
TATTGTCAAT
ATATTCAAGG
CAATCTGCCT
CCTCATCCTC
TTCATCCTCT
TCGTCTTGGT
AGCTTTTTAA
ATATGGCGCT
TCATAGAGTA
ATTCTGTAAA
GGTCCAATTC
TCGTTTTCAT
ACCTCGGTAT
AATCTTACCT
ATCACCTCAA
ATGGTTCGCT
GGGTTTATCG
GACCACTTCT
GCGCTCGGCC
CTTCCGGCTG
GCTGGTTTAT
TGCTGATAAA
TCTGGAGCCG
GTGAGCGTGG
GTCTCGCGGT
ATCATTGCAG
CACTGGGGCC
AGATGGTAAG
CCCTCCCGTA
TCGTAGTTAT
CTACACGACG
GGGAGTCAGG
CAACTATGGA
TGAACGAAAT
AGACAGATCG
CTGAGATAGG
TGCCTCACTG
ATTAAGCATT
GGTAACTGTC
AGACCAAGTT
TACTCATATA
TACTTTAGAT
TGATTTAAAA
CTTCATTTTT
AATTTAAAAG
GATCTAGGTG
AAGATCCTTT
TTGATAATCT
CATGACCAAA
ATCCCTTAAC
GTGAGTTTTC
GTTCCACTGA
GCGTCAGACC
CCGTAGAAAA
GATCAAAGGA
TCTTCTTGAG
ATCCTTTTTT
TCTGCGCGTA
ATCTGCTGCT
TGCAAACAAA
AAAACCACCG
CTACCAGCGG
TGGTTTGTTT
GCCGGATCAA
GAGCTACCAA
CTCTTTTTCC
GAAGGTAACT
GGCTTCAGCA
GAGCGCAGAT
ACCAAATACT
GTCCTTCTAG
TGTAGCCGTA
GTTAGGCCAC
CACTTCAAGA
ACTCTGTAGC
ACCGCCTACA
TACCTCGCTC
TGCTAATCCT
GTTACCAGTG
GCTGCTGCCA
GTGGCGATAA
GTCGTGTCTT
ACCGGGTTGG
ACTCAAGACG
ATAGTTACCG
GATAAGGCGC
AGCGGTCGGG
CTGAACGGGG
GGTTCGTGCA
CACAGCCCAG
CTTGGAGCGA
ACGACCTACA
CCGAACTGAG
ATACCTACAG
CGTGAGCATT
GAGAAAGCGC
CACGCTTCCC
GAAGGGAGAA
AGGCGGACAG
GTATCCGGTA
AGCGGCAGGG
TCGGAACAGG
AGAGCGCACG
AGGGAGCTTC
CAGGGGGAAA
CGCCTGGTAT
CTTTATAGTC
CTGTCGGGTT
TCGCCACCTC
TGACTTGAGC
GTCGATTTTT
GTGATGCTCG
TCAGGGGGGC
GGAGCCTATG
GAAAAACGCC
AGCAACGCGG
CCTTTTTACG
GTTCCTGGCC
TTTTGCTGGC
CTTTTGCTCA
CATGTTCTTT
CCTGCGTTAT
CCCCTGATTC
TGTGGATAAC
CGTATTACCG
CCTTTGAGTG
AGCTGATACC
GCTCGCCGCA
GCCGAACGAC
CGAGCGCAGC
GAGTCAGTGA
GCGAGGAAGC
GGAAGAGCGC
CCAATACGCA
AACCGCCTCT
CCCCGCGCGT
TGGCCGATTC
ATTAATGCAG