名称: | pDG1730 |
---|---|
别称: | |
基因长: | |
质粒宿主: | 枯草杆菌 |
质粒用途: | 蛋白表达 |
片段类型: | ORF |
片段物种: | |
原核抗性: | Amp |
筛选标记: | Ery,Spe |
荧光标记: | |
启动子: | |
复制子: | |
拷贝数: | |
感受态: | Stbl3 |
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。
AACAAAATTC
TCCAGTCTTC
ACATCGGTTT
GAAAGGAGGA
AGCGGAAGAA
TGAAGTAAGA
GGGATTTTTG
ACTCCGAAGT
AAGTCTTCAA
AAAATCAAAT
AAGGAGTGTC
AAGAATGTTT
GCAAAACGAT
TCAAAACCTC
TTTACTGCCG
TTATTCGCTG
GATTTTTATT
GCTGTTTCAT
TTGGTTCTGG
CAGGACCGGC
GGCTGCGAGT
GCTGAAACGG
CGAACAAATC
GAATGAGCTT
ACAGCACCGT
CGATCAAAAG
CGGAACCATT
CTTCATGCAT
GGAATTGGTC
GTTCAATACG
TTAAAACACA
ATATGAAGGA
TATTCATGAT
GCAGGATATA
CAGCCATTCA
GACATCTCCG
ATTAACCAAG
TAAAGGAAGG
GAATCAAGGA
GATAAAAGCA
TGTCGAACTG
GTACTGGCTG
TATCAGCCGA
CATCGTATCA
AATTGGCAAC
CGTTACTTAG
GTACTGAACA
AGAATTTAAA
GAAATGTGTG
CAGCCGCTGA
AGAATATGGC
ATAAAGGTCA
TTGTTGACGC
GGTCATCAAT
CATACCACCA
GTGATTATGC
CGCGATTTCC
AATGAGGTTA
AGAGTATTCC
AAACTGGACA
CATGGAAACA
CACAAATTAA
AAACTGGTCT
GATCGGATCC
TAGAAGCTTA
TCGAATTCCT
GCAGCCCTGG
CGAATGGCGA
TTTTCGTTCG
TGAATACATG
TTATAATAAC
TATAACTAAT
AACGTAACGT
GACTGGCAAG
AGATATTTTT
AAAACAATGA
ATAGGTTTAC
ACTTACTTTA
GTTTTATGGA
AATGAAAGAT
CATATCATAT
ATAATCTAGA
ATAAAATTAA
CTAAAATAAT
TATTATCTAG
ATAAAAAATT
TAGAAGCCAA
TGAAATCTAT
AAATAAACTA
AATTAAGTTT
ATTTAATTAA
CAACTATGGA
TATAAAATAG
GTACTAATCA
AAATAGTGAG
GAGGATATAT
TTGAATACAT
ACGAACAAAT
TAATAAAGTG
AAAAAAATAC
TTCGGAAACA
TTTAAAAAAT
AACCTTATTG
GTACTTACAT
GTTTGGATCA
GGAGTTGAGA
GTGGACTAAA
ACCAAATAGT
GATCTTGACT
TTTTAGTCGT
CGTATCTGAA
CCATTGACAG
ATCAAAGTAA
AGAAATACTT
ATACAAAAAA
TTAGACCTAT
TTCAAAAAAA
ATAGGAGATA
AAAGCAACTT
ACGATATATT
GAATTAACAA
TTATTATTCA
GCAAGAAATG
GTACCGTGGA
ATCATCCTCC
CAAACAAGAA
TTTATTTATG
GAGAATGGTT
ACAAGAGCTT
TATGAACAAG
GATACATTCC
TCAGAAGGAA
TTAAATTCAG
ATTTAACCAT
AATGCTTTAC
CAAGCAAAAC
GAAAAAATAA
AAGAATATAC
GGAAATTATG
ACTTAGAGGA
ATTACTACCT
GATATTCCAT
TTTCTGATGT
GAGAAGAGCC
ATTATGGATT
CGTCAGAGGA
ATTAATAGAT
AATTATCAGG
ATGATGAAAC
CAACTCTATA
TTAACTTTAT
GCCGTATGAT
TTTAACTATG
GACACGGGTA
AAATCATACC
AAAAGATATT
GCGGGAAATG
CAGTGGCTGA
ATCTTCTCCA
TTAGAACATA
GGGAGAGAAT
TTTGTTAGCA
GTTCGTAGTT
ATCTTGGAGA
GAATATTGAA
TGGACTAATG
AAAATGTAAA
TTTAACTATA
AACTATTTAA
ATAACAGATT
AAAAAAATTA
TAAAAAAATT
GAAAAAATGG
TGGAAACACT
TTTTTCAATT
TTTTTGTTTT
ATTATTTAAT
ATTTGGGAAA
TATTCATTCT
AATTGGTAAT
CAGATTTTAG
AAAACAATAA
ACCCTTGCAT
AGGGGGATCT
CGACATGGAT
GAGCGATGAT
GATATCCGTT
TAGGCTGGGC
GGTGATAGCT
TCTCGTTCAG
GCAGTACGCC
TCTTTTCTTT
TCCAGACCTG
AGGGAGGCGG
AAATGGTGTG
AGGTTCCCGG
GGAAAAGCCA
AATAGGCGAT
CGCGGGAGTG
CTTTATTTGA
AGATCAGGCT
ATCACTGCGG
TCAATAGATT
TCACAATGTG
ATGGCTGGAC
AGCCTGAGGA
ACTCTCGAAC
CCGAATGGAA
ACAACCAGAT
ATTTATGAAT
CAGCGCGGCT
CACATGGCGT
TGTGCTGGCA
AATGCAGGTT
CATCCTCTGT
CTCTATCAAT
ACGGCAACAA
AATTGCCTGA
TGGCAGGTAT
GACAATAAAG
CTGGAGCGGG
TTCATTTCAA
GTGAACGATG
GTAAACTGAC
AGGCACGATC
AATGCCAGGT
CTGTAGCTGT
GCTTTATCCT
GATGATATTG
CAAAAGCGCC
TCATGTTTTC
CTTGAGAATT
ACAAAACAGG
TGTAACACAT
TCTTTCAATG
ATCAACTGAC
GATTACCTTG
CGTGCAGATG
CGAATACAAC
AAAAGCCGTT
TATCAAATCA
ATAATGGACC
AGACGACAGG
CGTTTAAGGA
TGGAGATCAA
TTCACAATCG
GAAAAGGAGA
TCCAATTTGG
CAAAACATAC
ACCATCATGT
TAAAAGGAAC
GAACAGTGAT
GGTGTAACGA
GGACCGAGAA
ATACAGTTTT
GTTAAAAGAG
ATCCAGCGTC
GGCCAAAACC
ATCGGCTATC
AAAATCCGAA
TCATTGGAGC
CAGGTAAATG
CTTATATCTA
TAAACATGAT
GGGAGCCGAG
TAATTGAATT
GACCGGATCT
TGGCCTGGAA
AACCAATGAC
TAAAAATGCA
GACGGAATTT
ACACGCTGAC
GCTGCCTGCG
GACACGGATA
CAACCAACGC
AAAAGTGATT
TTTAATAATG
GCAGCGCCCA
AGTGCCCGGT
CAGAATCAGC
CTGGCTTTGA
TTACGTGCTA
AATGGTTTAT
ATAATGACTC
GGGCTTAAGC
GGTTCTCTTC
CCCATTGAGG
GCAAGGCTAG
ACGGGACTTA
CCGAAAGAAA
CCATCAATGA
TGGTTTCTTT
TTTGTTCATA
AATCAGACAA
AACTTTTCTC
TTGCAAAAGT
TTGTGAAGTG
TTGCACAATA
TAAATGTGAA
ATACTTCACA
AACAAAAAGA
CATCAAAGAG
AAACATACCC
TGCAAGGATG
CTGATATTGT
CTGCATTTGC
GCCGGAGCAA
ACCAAAAACC
TGGTGAGACA
CGCCTTGAAT
TAGTAGAAAA
GAACTTGAAG
ATTTTCAAAG
GCATCGTTAG
TGAAGTCATG
GCGAGCGGAT
TTGACGGCAT
TTTCTTAGTC
GCGACGCGAG
GCTGGATGGC
CTTCCCCATT
ATGATTCTTC
TCGCTTCCGG
CGGCATCGGG
ATGCCCGCGT
TGCAGGCCAT
GCTGTCCAGG
CAGGTAGATG
ACGACCATCA
GGGACAGCTT
CAAGGATCGC
TCGCGGCTCT
TACCAGCCTA
ACTTCGATCA
CTGGACCGCT
GATCGTCACG
GCGATTTATG
CCGCCTCGGC
GAGCACATGG
AACGGGTTGG
CATGGATTGT
AGGCGCCGCC
CTATACCTTG
TCTGCCTCCC
CGCGTTGCGT
CGCGGTGCAT
GGAGCCGGGC
CACCTACTGA
AGTGGATTTC
TTTAAGAGCT
CCTTTAACTT
CCTCACCAGT
AGTTGTATCG
GTACCATAAG
TAGAAGCAGC
AACCCAAGTA
GCTTTACCAG
CATCCGGTTC
AACCAGCATA
GTAAGAATCT
TACTGGACAT
CGGCAGTTCT
TCGAACAGTG
CGCCAACTAC
CAGCTCTTTC
TGCAGTTCAT
TCAGGGCACC
GGAGAACCTG
CGTGCAATCC
ATCTTGTTCA
ATCATGCGAA
ACGATCCTCA
TCCTGTCTCT
TGATCCATGG
ATTACGCGTT
AACCCGGGCC
CGCGGATGCA
TATGATCAGA
TCCTTTAACT
CTGGCAACCC
TCAAAATTGA
ATGAGACATG
CTACACCTCC
GGATAATAAA
TATATATAAA
CGTATATAGA
TTTCATAAAG
TCTAACACAC
TAGACTTATT
TACTTCGTAA
TTAAGTCGTT
AAACCGTGTG
CTCTACGACC
AAAACTATAA
AACCTTTAAG
AACTTTCTTT
TTTTACAAGA
AAAAAGAAAT
TAGATAAATC
TCTCATATCT
TTTATTCAAT
AATCGCATCC
GATTGCAGTA
TAAATTTAAC
GATCACTCAT
CATGTTCATA
TTTATCAGAG
CTCGTGCTAT
AATTATACTA
ATTTTATAAG
GAGGAAAAAA
TATGGGCATT
TTTAGTATTT
TTGTAATCAG
CACAGTTCAT
TATCAACCAA
ACAAAAAATA
AGTGGTTATA
ATGAATCGTT
AATAAGCAAA
ATTCATATAA
CCAAATTAAA
GAGGGTTATA
ATGAACGAGA
AAAATATAAA
ACACAGTCAA
AACTTTATTA
CTTCAAAACA
TAATATAGAT
AAAATAATGA
CAAATATAAG
ATTAAATGAA
CATGATAATA
TCTTTGAAAT
CGGCTCAGGA
AAAGGCCATT
TTACCCTTGA
ATTAGTAAAG
AGGTGTAATT
TCGTAACTGC
CATTGAAATA
GACCATAAAT
TATGCAAAAC
TACAGAAAAT
AAACTTGTTG
ATCACGATAA
TTTCCAAGTT
TTAAACAAGG
ATATATTGCA
GTTTAAATTT
CCTAAAAACC
AATCCTATAA
AATATATGGT
AATATACCTT
ATAACATAAG
TACGGATATA
ATACGCAAAA
TTGTTTTTGA
TAGTATAGCT
AATGAGATTT
ATTTAATCGT
GGAATACGGG
TTTGCTAAAA
GATTATTAAA
TACAAAACGC
TCATTGGCAT
TACTTTTAAT
GGCAGAAGTT
GATATTTCTA
TATTAAGTAT
GGTTCCAAGA
GAATATTTTC
ATCCTAAACC
TAAAGTGAAT
AGCTCACTTA
TCAGATTAAG
TAGAAAAAAA
TCAAGAATAT
CACACAAAGA
TAAACAAAAG
TATAATTATT
TCGTTATGAA
ATGGGTTAAC
AAAGAATACA
AGAAAATATT
TACAAAAAAT
CAATTTAACA
ATTCCTTAAA
ACATGCAGGA
ATTGACGATT
TAAACAATAT
TAGCTTTGAA
CAATTCTTAT
CTCTTTTCAA
TAGCTATAAA
TTATTTAATA
AGTAAGTTAA
GGGATGCATA
AACTGCATCC
CTTAACTTGT
TTTTCGTGTG
CCTATTTTTT
GTGAATCGAT
TATGTCTTTT
GCGCAGTCGG
CTTAAACCAG
TTTTCGCTGG
TGCGAAAAAA
GAGTGTCTTG
TGACACCTAA
ATTCAAAATC
TATCGGTCAG
ATTTATACCG
ATTTGATTTT
ATATATTCTT
GAATAACATA
CGCCGAGTTA
TCACATAAAA
GCGGGAACCA
ATCATCAAAT
TTAAACTTCA
TTGCATAATC
CATTAAACTC
TTAAATTCTA
CGATTCCTTG
TTCATCAATA
AACTCAATCA
TTTCTTTAAT
TAATTTATAT
CTATCTGTTG
TTGTTTTCTT
TAATAATTCA
TCAACATCTA
CACCGCCATA
AACTATCATA
TCTTCTTTTT
GATATTTAAA
TTTATTAGGA
TCTTAAGGCC
TAGGTCTAGA
GTCTTTGTTT
TGACGCCATT
AGCGTACGTA
ACAATCCTCG
TTAAAGGACA
AGGACCTGAG
CGGAAGTGTA
TCGTACAGTA
GACGGAGTAT
ACTAGTATAG
TCTATAGTCC
GTGGAATTAT
TATATTTATC
TCCGACGATA
TTCTCATCAG
TGAAATCCAG
CTGGAGTTCT
TTAGCAAATT
TTTTTATTAG
CTGAACTTAG
TATTAGTGGC
CATACTCCTC
CAATCCAAAG
CTATTTAGAA
AGATTACTAT
ATCCTCAAAC
AGGCGGTAAC
CGGCCTCTTC
ATCGGGAATG
CGCGCGACCT
TCAGCATCGC
CGGCATGTCC
CCCTGGCGGA
CGGGAAGTAT
CCAGCTCGAG
GTCGGGCCGC
GTTGCTGGCG
TTTTTCCATA
GGCTCCGCCC
CCCTGACGAG
CATCACAAAA
ATCGACGCTC
AAGTCAGAGG
TGGCGAAACC
CGACAGGACT
ATAAAGATAC
CAGGCGTTTC
CCCCTGGAAG
CTCCCTCGTG
CGCTCTCCTG
TTCCGACCCT
GCCGCTTACC
GGATACCTGT
CCGCCTTTCT
CCCTTCGGGA
AGCGTGGCGC
TTTCTCATAG
CTCACGCTGT
AGGTATCTCA
GTTCGGTGTA
GGTCGTTCGC
TCCAAGCTGG
GCTGTGTGCA
CGAACCCCCC
GTTCAGCCCG
ACCGCTGCGC
CTTATCCGGT
AACTATCGTC
TTGAGTCCAA
CCCGGTAAGA
CACGACTTAT
CGCCACTGGC
AGCAGCCACT
GGTAACAGGA
TTAGCAGAGC
GAGGTATGTA
GGCGGTGCTA
CAGAGTTCTT
GAAGTGGTGG
CCTAACTACG
GCTACACTAG
AAGGACAGTA
TTTGGTATCT
GCGCTCTGCT
GAAGCCAGTT
ACCTTCGGAA
AAAGAGTTGA
TAGCTCTTGA
TCCGGCAAAC
AAACCACCGC
TGGTAGCGGT
GGTTTTTTTG
TTTGCAAGCA
GCAGATTACG
CGCAGAAAAA
AAGGATCTCA
AGAAGATCCT
TTGATCTTTT
CTACGGGGTC
TGACGCTCAG
TGGAACGAAA
ACTCACGTTA
AGGGATTTTG
GTCATGAGAT
TATCAAAAAG
GATCTTCACC
TAGATCCTTT
TAAATTAAAA
ATGAAGTTTT
AAATCAATCT
AAAGTATATA
TGAGTAAACT
TGGTCTGACA
GTTACCAATG
CTTAATCAGT
GAGGCACCTA
TCTCAGCGAT
CTGTCTATTT
CGTTCATCCA
TAGTTGCCTG
ACTCCCCGTC
GTGTAGATAA
CTACGATACG
GGAGGGCTTA
CCATCTGGCC
CCAGTGCTGC
AATGATACCG
CGAGACCCAC
GCTCACCGGC
TCCAGATTTA
TCAGCAATAA
ACCAGCCAGC
CGGAAGGGCC
GAGCGCAGAA
GTGGTCCTGC
AACTTTATCC
GCCTCCATCC
AGTCTATTAA
TTGTTGCCGG
GAAGCTAGAG
TAAGTAGTTC
GCCAGTTAAT
AGTTTGCGCA
ACGTTGTTGC
CATTGCTGCA
GGCATCGTGG
TGTCACGCTC
GTCGTTTGGT
ATGGCTTCAT
TCAGCTCCGG
TTCCCAACGA
TCAAGGCGAG
TTACATGATC
CCCCATGTTG
TGCAAAAAAG
CGGTTAGCTC
CTTCGGTCCT
CCGATCGTTG
TCAGAAGTAA
GTTGGCCGCA
GTGTTATCAC
TCATGGTTAT
GGCAGCACTG
CATAATTCTC
TTACTGTCAT
GCCATCCGTA
AGATGCTTTT
CTGTGACTGG
TGAGTACTCA
ACCAAGTCAT
TCTGAGAATA
GTGTATGCGG
CGACCGAGTT
GCTCTTGCCC
GGCGTCAACA
CGGGATAATA
CCGCGCCACA
TAGCAGAACT
TTAAAAGTGC
TCATCATTGG
AAAACGTTCT
TCGGGGCGAA
AACTCTCAAG
GATCTTACCG
CTGTTGAGAT
CCAGTTCGAT
GTAACCCACT
CGTGCACCCA
ACTGATCTTC
AGCATCTTTT
ACTTTCACCA
GCGTTTCTGG
GTGAGCAAAA
ACAGGAAGGC
AAAATGCCGC
AAAAAAGGGA
ATAAGGGCGA
CACGGAAATG
TTGAATACTC
ATACTCTTCC
TTTTTCAATA
TTATTGAAGC
ATTTATCAGG
GTTATTGTCT
CATGAGCGGA
TACATATTTG
AATGTATTTA
GAAAAATAAA
CAAATAGGGG
TTCCGCGCAC
ATTTCCCCGA
AAAGTGCCAC
CTGACGTCTA
AGAAACCATT
ATTATCATGA
CATTAACCTA
TAAAAATAGG
CGTATCACGA
GGCCCTTTCG
TCTTCAAGAA
TT