名称: | BB3cH_pGAP_23*_pPFK300_Cas9 |
---|---|
别称: | Plasmid#104912 |
基因长: | |
质粒宿主: | 酵母菌 |
质粒用途: | 基因编辑 |
片段类型: | CRISPR |
片段物种: | |
原核抗性: | Hyg |
筛选标记: | Hyg |
荧光标记: | |
启动子: | GAP |
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。
CGCTGTGAGC
AAAAGGCCAG
CAAAAGGCCA
GGAACCGTAA
AAAGGCCGCG
TTGCTGGCGT
TTTTCCATAG
GCTCCGCCCC
CCTGACGAGC
ATCACAAAAA
TCGACGCTCA
AGTCAGAGGT
GGCGAAACCC
GACAGGACTA
TAAAGATACC
AGGCGTTTCC
CCCTGGAAGC
TCCCTCGTGC
GCTCTCCTGT
TCCGACCCTG
CCGCTTACCG
GATACCTGTC
CGCCTTTCTC
CCTTCGGGAA
GCGTGGCGCT
TTCTCATAGC
TCACGCTGTA
GGTATCTCAG
TTCGGTGTAG
GTCGTTCGCT
CCAAGCTGGG
CTGTGTGCAC
GAACCCCCCG
TTCAGCCCGA
CCGCTGCGCC
TTATCCGGTA
ACTATCGTCT
TGAGTCCAAC
CCGGTAAGAC
ACGACTTATC
GCCACTGGCA
GCAGCCACTG
GTAACAGGAT
TAGCAGAGCG
AGGTATGTAG
GCGGTGCTAC
AGAGTTCTTG
AAGTGGTGGC
CTAACTACGG
CTACACTAGA
AGAACAGTAT
TTGGTATCTG
CGCTCTGCTG
AAGCCAGTTA
CCTTCGGAAA
AAGAGTTGGT
AGCTCTTGAT
CCGGCAAACA
AACCACCGCT
GGTAGCGGTG
GTTTTTTTGT
TTGCAAGCAG
CAGATTACGC
GCAGAAAAAA
AGGATCTCAA
GAAGATCCTT
TGATCTTTTC
TACGGGGTGG
AGCGCAAAAA
ACCCCGCCCC
TGACAGGGCG
GGGTTTTTTC
GCGATCGGAG
GATCCTTTTT
TGTAGAAATG
TCTTGGTGTC
CTCGTCCAAT
CAGGTAGCCA
TCTCTGAAAT
ATCTGGCTCC
GTTGCAACTC
CGAACGACCT
GCTGGCAACG
TAAAATTCTC
CGGGGTAAAA
CTTAAATGTG
GAGTAATGGA
ACCAGAAACG
TCTCTTCCCT
TCTCTCTCCT
TCCACCGCCC
GTTACCGTCC
CTAGGAAATT
TTACTCTGCT
GGAGAGCTTC
TTCTACGGCC
CCCTTGCAGC
AATGCTCTTC
CCAGCATTAC
GTTGCGGGTA
AAACGGAGGT
CGTGTACCCG
ACCTAGCAGC
CCAGGGATGG
AAAAGTCCCG
GCCGTCGCTG
GCAATAATAG
CGGGCGGACG
CATGTCATGA
GATTATTGGA
AACCACCAGA
ATCGAATATA
AAAGGCGAAC
ACCTTTCCCA
ATTTTGGTTT
CTCCTGACCC
AAAGACTTTA
AATTTAATTT
ATTTGTCCCT
ATTTCAATCA
ATTGAACAAC
TATCAAAACA
CCATGGCGTC
TTCGGATCCC
TCGAGGAAGA
CGCGCTTTTA
GTGTACATCT
GATAATATAG
TACTACCACG
TATGATAATG
TAGAGAATAG
GCTTCCTTGT
CGAGTGTGTT
TGCAGTTTTC
TTGAGTTTCA
AGGTTTAAAT
GCTGGTATAT
TAGTTCATCG
AAGGTTTCAG
CCAATAGCAC
CTTAAATCAA
TCAAACTAAT
TCGACTCTTA
CGAAAGAGCC
TACTGTGTTT
AGTATCGAAG
TCGTTTACCT
TTCATGTTGA
ATAGCTTCCT
CTCTGACCCT
AACATTTCAA
GATCCTCCTA
AAGTTACCCG
GATTGTGAAA
TTCTAATGAT
CCACCTGCCC
AATGCATTTT
TTCTTTATTC
AGTTTACCTT
TTTTACCTAA
TATACGAGCT
TGTTAAAGTA
AGTGGCACTG
CAATACTAGG
CTTATTGTTG
ATATTATGAT
GAATCGTTTT
CACAAACTTG
ATTTCCTGTG
AACTCACCAT
GTACTAAGGA
AAAAAACATG
CATCACCATC
TGAATATTTG
ACCGCTCCGG
GGAGCCTGCC
ATTAGGCAAA
CACCAACTAG
GAACCTACAC
CAAGTTATGA
AGGAGCTTGG
AAAGGATTAT
AAAAAATGCG
ATTACCTAAT
ACACACACGA
TGCCTGAGCG
GTTAATGTGA
AATTTCCCGG
CCATCGATCG
CCCCACAATC
AGACTATTTT
CGCTCATACA
ATAAAAAGTC
TCCTGTTTCC
ACCCGAGTAT
CACCAGAACA
AAGTAACAAA
TACAAAACCA
TGGACAAGAA
GTACTCCATT
GGGCTCGATA
TCGGCACAAA
CAGCGTCGGT
TGGGCCGTCA
TTACGGACGA
GTACAAGGTG
CCGAGCAAAA
AATTCAAAGT
TCTGGGCAAT
ACCGATCGCC
ACAGCATAAA
GAAGAACCTC
ATTGGCGCCC
TCCTGTTCGA
CTCCGGGGAG
ACGGCCGAAG
CCACGCGGCT
CAAAAGAACA
GCACGGCGCA
GATATACCCG
CAGAAAGAAT
CGGATCTGCT
ACCTGCAGGA
GATCTTTAGT
AATGAGATGG
CTAAGGTGGA
TGACTCTTTC
TTCCATAGGC
TGGAGGAGTC
CTTTTTGGTG
GAGGAGGATA
AAAAGCACGA
GCGCCACCCA
ATCTTTGGCA
ATATCGTGGA
CGAGGTGGCG
TACCATGAAA
AGTACCCAAC
CATATATCAT
CTGAGGAAGA
AGCTTGTAGA
CAGTACTGAT
AAGGCTGACT
TGCGGTTGAT
CTATCTCGCG
CTGGCGCATA
TGATCAAATT
TCGGGGACAC
TTCCTCATCG
AGGGGGACCT
GAACCCAGAC
AACAGCGATG
TCGACAAACT
CTTTATCCAA
CTGGTTCAGA
CTTACAATCA
GCTTTTCGAA
GAGAACCCGA
TCAACGCATC
CGGAGTTGAC
GCCAAAGCAA
TCCTGAGCGC
TAGGCTGTCC
AAATCCCGGC
GGCTCGAAAA
CCTCATCGCA
CAGCTCCCTG
GGGAGAAGAA
GAACGGCCTG
TTTGGTAATC
TTATCGCCCT
GTCACTCGGG
CTGACCCCCA
ACTTTAAATC
TAACTTCGAC
CTGGCCGAAG
ATGCCAAGCT
TCAACTGAGC
AAAGACACCT
ACGATGATGA
TCTCGACAAT
CTGCTGGCCC
AGATCGGCGA
CCAGTACGCA
GACCTTTTTT
TGGCGGCAAA
GAACCTGTCA
GACGCCATTC
TGCTGAGTGA
TATTCTGCGA
GTGAACACGG
AGATCACCAA
AGCTCCGCTG
AGCGCTAGTA
TGATCAAGCG
CTATGATGAG
CACCACCAAG
ACTTGACTTT
GCTGAAGGCC
CTTGTCAGAC
AGCAACTGCC
TGAGAAGTAC
AAGGAAATTT
TCTTCGATCA
GTCTAAAAAT
GGCTACGCCG
GATACATTGA
CGGCGGAGCA
AGCCAGGAGG
AATTTTACAA
ATTTATTAAG
CCCATCTTGG
AAAAAATGGA
CGGCACCGAG
GAGCTGCTGG
TAAAGCTTAA
CAGAGAAGAT
CTGTTGCGCA
AACAGCGCAC
TTTCGACAAT
GGAAGCATCC
CCCACCAGAT
TCACCTGGGC
GAACTGCACG
CTATCCTCAG
GCGGCAAGAG
GATTTCTACC
CCTTTTTGAA
AGATAACAGG
GAAAAGATTG
AGAAAATCCT
CACATTTCGG
ATACCCTACT
ATGTAGGCCC
CCTCGCCCGG
GGAAATTCCA
GATTCGCGTG
GATGACTCGC
AAATCAGAAG
AGACTATCAC
TCCCTGGAAC
TTCGAGGAAG
TCGTGGATAA
GGGGGCCTCT
GCCCAGTCCT
TCATCGAAAG
GATGACTAAC
TTTGATAAAA
ATCTGCCTAA
CGAAAAGGTG
CTTCCTAAAC
ACTCTCTGCT
GTACGAGTAC
TTCACAGTTT
ATAACGAGCT
CACCAAGGTC
AAATACGTCA
CAGAAGGGAT
GAGAAAGCCA
GCATTCCTGT
CTGGAGAGCA
GAAGAAAGCT
ATCGTGGACC
TCCTCTTCAA
GACGAACCGG
AAAGTTACCG
TGAAACAGCT
CAAAGAGGAC
TATTTCAAAA
AGATTGAATG
TTTCGACTCT
GTTGAAATCA
GCGGAGTGGA
GGATCGCTTC
AACGCATCCC
TGGGAACGTA
TCACGATCTC
CTGAAAATCA
TTAAAGACAA
GGACTTCCTG
GACAATGAGG
AGAACGAGGA
CATTCTTGAG
GACATTGTCC
TCACCCTTAC
GTTGTTTGAA
GATAGGGAGA
TGATTGAAGA
ACGCTTGAAA
ACTTACGCTC
ATCTCTTCGA
CGACAAAGTC
ATGAAACAGC
TCAAGAGGCG
CCGATATACA
GGATGGGGGC
GGCTGTCAAG
AAAACTGATC
AATGGGATCC
GAGACAAGCA
GAGTGGAAAG
ACAATCCTGG
ATTTTCTTAA
GTCCGATGGA
TTTGCCAACC
GGAACTTCAT
GCAGTTGATC
CATGATGACT
CTCTCACCTT
TAAGGAGGAC
ATCCAGAAAG
CACAAGTTTC
TGGCCAGGGG
GACAGCCTTC
ACGAGCACAT
CGCTAATCTT
GCAGGTAGCC
CAGCTATCAA
AAAGGGAATA
CTGCAGACCG
TTAAGGTCGT
GGATGAACTC
GTCAAAGTAA
TGGGAAGGCA
TAAGCCCGAG
AATATCGTTA
TCGAGATGGC
CCGAGAGAAC
CAAACTACCC
AGAAGGGACA
GAAGAACAGT
AGGGAAAGGA
TGAAGAGGAT
TGAAGAGGGT
ATAAAAGAAC
TGGGGTCCCA
AATCCTTAAG
GAACACCCAG
TTGAAAACAC
CCAGCTTCAG
AATGAGAAGC
TCTACCTGTA
CTACCTGCAG
AACGGCAGGG
ACATGTACGT
GGATCAGGAA
CTGGACATCA
ATCGGCTCTC
CGACTACGAC
GTGGATCATA
TCGTGCCCCA
GTCTTTTCTC
AAAGATGATT
CTATTGATAA
TAAAGTGTTG
ACAAGATCCG
ATAAAAATAG
AGGGAAGAGT
GATAACGTCC
CCTCAGAAGA
AGTTGTCAAG
AAAATGAAAA
ATTATTGGCG
GCAGCTGCTG
AACGCCAAAC
TGATCACACA
ACGGAAGTTC
GATAATCTGA
CTAAGGCTGA
ACGAGGTGGC
CTGTCTGAGT
TGGATAAAGC
CGGCTTCATC
AAAAGGCAGC
TTGTTGAGAC
ACGCCAGATC
ACCAAGCACG
TGGCCCAAAT
TCTCGATTCA
CGCATGAACA
CCAAGTACGA
TGAAAATGAC
AAACTGATTC
GAGAGGTGAA
AGTTATTACT
CTGAAGTCTA
AGCTCGTCTC
AGATTTCAGA
AAGGACTTTC
AGTTTTATAA
GGTGAGAGAG
ATCAACAATT
ACCACCATGC
GCATGATGCC
TACCTGAATG
CAGTGGTAGG
CACTGCACTT
ATCAAAAAAT
ATCCCAAGCT
TGAATCTGAA
TTTGTTTACG
GAGACTATAA
AGTGTACGAT
GTTAGGAAAA
TGATCGCAAA
GTCTGAGCAG
GAAATAGGCA
AGGCCACCGC
TAAGTACTTC
TTTTACAGCA
ATATTATGAA
TTTTTTCAAG
ACCGAGATTA
CACTGGCCAA
TGGAGAGATT
CGGAAGCGAC
CACTTATCGA
AACAAACGGA
GAAACAGGAG
AAATCGTGTG
GGACAAGGGT
AGGGATTTCG
CGACAGTCCG
GAAGGTCCTG
TCCATGCCGC
AGGTGAACAT
CGTTAAAAAG
ACCGAAGTAC
AGACCGGAGG
CTTCTCCAAG
GAAAGTATCC
TCCCGAAAAG
GAACAGCGAC
AAGCTGATCG
CACGCAAAAA
AGATTGGGAC
CCCAAGAAAT
ACGGCGGATT
CGATTCTCCT
ACAGTCGCTT
ACAGTGTACT
GGTTGTGGCC
AAAGTGGAGA
AAGGGAAGTC
TAAAAAACTC
AAAAGCGTCA
AGGAACTGCT
GGGCATCACA
ATCATGGAGC
GATCAAGCTT
CGAAAAAAAC
CCCATCGACT
TTCTCGAGGC
GAAAGGATAT
AAAGAGGTCA
AAAAAGACCT
CATCATTAAG
CTTCCCAAGT
ACTCTCTCTT
TGAGCTTGAA
AACGGCCGGA
AACGAATGCT
CGCTAGTGCG
GGCGAGCTGC
AGAAAGGTAA
CGAGCTGGCA
CTGCCCTCTA
AATACGTTAA
TTTCTTGTAT
CTGGCCAGCC
ACTATGAAAA
GCTCAAAGGG
TCCCCCGAAG
ATAATGAGCA
GAAGCAGCTG
TTCGTGGAAC
AACACAAACA
CTACCTTGAT
GAGATCATCG
AGCAAATAAG
CGAATTCTCC
AAAAGAGTGA
TCCTCGCCGA
CGCTAACCTC
GATAAGGTGC
TTTCTGCTTA
CAATAAGCAC
AGGGATAAGC
CCATCAGGGA
GCAGGCAGAA
AACATTATCC
ACTTGTTTAC
TCTGACCAAC
TTGGGCGCGC
CTGCAGCCTT
CAAGTACTTC
GACACCACCA
TAGACAGAAA
GCGGTACACC
TCTACAAAGG
AGGTCCTGGA
CGCCACACTG
ATTCATCAGT
CAATTACGGG
GCTCTATGAA
ACAAGAATCG
ACCTCTCTCA
GCTCGGTGGA
GACAGCAGGG
CTGACCCCAA
GAAGAAGAGG
AAGGTGTGAG
CTTCGTCCGA
CGGCGGCCCG
ACGGGTCCGA
GGCCTCGGAG
ATCCGTCCCC
CTTTTCCTTT
GTCGATATCA
TGTAATTAGT
TATGTCACGC
TTACATTCAC
GCCCTCCCCC
CACATCCGCT
CTAACCGAAA
AGGAAGGAGT
TAGACAACCT
GAAGTCTAGG
TCCCTATTTA
TTTTTTTATA
GTTATGTTAG
TATTAAGAAC
GTTATTTATA
TTTCAAATTT
TTCTTTTTTT
TCTGTACAGA
CGCGTGTACG
CATGTAACAT
TATACTGAAA
ACCTTGCTTG
AGAAGGTTTT
GGGACGCTCG
AAGGCTTTAA
TTTGCAAGCT
GGCGCTAATT
CATGGACATG
GAGGCCCAGA
ATACCCTCCT
TGACAGTCTT
GACGTGCGCA
GCTCAGGGGC
ATGATGTGAC
TGTCGCCCGT
ACATTTAGCC
CATACATCCC
CATGTATAAT
CATTTGCATC
CATACATTTT
GATGGCCGCA
CGGCGCGAAG
CAAAAATTAC
GGCTCCTCGC
TGCAGACGTG
CGAGCAGGGA
AACGCTCCCC
TCACAGACGC
GTTGAATTGT
CCCCACGCCG
CGCCCCTGTA
GAGAAATATA
AAAGGTTAGG
ATTTGCCACT
GAGGTTCTTC
TTTCATATAC
TTCCTTTTAA
AATCTTGCTA
GGATACAGTT
CTCACATCAC
ATCCGAACAT
AAACAACCAT
GGGTAAAAAG
CCTGAACTCA
CCGCGACGTC
TGTCGAGAAG
TTTCTGATCG
AAAAGTTCGA
CAGCGTCTCC
GACCTGATGC
AGCTCTCGGA
GGGCGAAGAA
TCTCGTGCTT
TCAGCTTCGA
TTTAGGAGGG
CGTGGATATG
TCCTGCGGGT
AAATAGCTGC
GCCGATGGTT
TCTACAAAGA
TCGTTATGTT
TATCGGCACT
TTGCATCGGC
CGCGCTCCCG
ATTCCGGAAG
TGCTTGACAT
TGGGGAATTC
AGCGAGAGCC
TGACCTATTG
CATCTCCCGC
CGTGCACAGG
GTGTCACGTT
GCAAGACCTC
CCTGAAACCG
AACTGCCCGC
TGTTCTGCAG
CCGGTCGCGG
AGGCCATGGA
TGCGATCGCT
GCGGCCGATC
TTAGCCAGAC
GAGCGGGTTC
GGCCCATTCG
GACCGCAAGG
AATCGGTCAA
TACACTACAT
GGCGTGATTT
CATATGCGCG
ATTGCTGATC
CCCATGTGTA
TCACTGGCAA
ACTGTGATGG
ACGACACCGT
CAGTGCGTCC
GTCGCGCAGG
CTCTCGATGA
GCTGATGCTT
TGGGCCGAGG
ACTGCCCCGA
AGTCCGGCAC
CTCGTGCACG
CGGATTTCGG
CTCCAACAAT
GTCCTGACGG
ACAATGGCCG
CATAACAGCG
GTCATTGACT
GGAGCGAGGC
GATGTTCGGG
GATTCCCAAT
ACGAGGTCGC
CAACATCTTC
TTCTGGAGGC
CGTGGTTGGC
TTGTATGGAG
CAGCAGACGC
GCTACTTCGA
GCGGAGGCAT
CCGGAGCTTG
CAGGATCGCC
GCGGCTCCGG
GCGTATATGC
TCCGCATTGG
TCTTGACCAA
CTCTATCAGA
GCTTGGTTGA
CGGCAATTTC
GATGATGCAG
CTTGGGCGCA
GGGTCGATGC
GACGCAATCG
TCCGATCCGG
AGCCGGGACT
GTCGGGCGTA
CACAAATCGC
CCGCAGAAGC
GCGGCCGTCT
GGACCGATGG
CTGTGTAGAA
GTACTCGCCG
ATAGTGGAAA
CCGACGCCCC
AGCACTCGTC
CGAGGGCAAA
GGAATAATCA
GTACTGACAA
TAAAAAGATT
CTTGTTTTCA
AGAACTTGTC
ATTTGTATAG
TTTTTTTATA
TTGTAGTTGT
TCTATTTTAA
TCAAATGTTA
GCGTGATTTA
TATTTTTTTT
CGCCTCGACA
TCATCTGCCC
AGATGCGAAG
TTAAGTGCGC
AGAAAGTAAT
ATCATGCGTC
AATCGTATGT
GAATGCTGGT
CGCTATACTG
GCTTCCTTTT
CATCACGTGC
TATAAAAATA
ATTATAATTT
AAATTTTTTA
ATATAAATAT
ATAAATTAAA
AATAGAAAGT
AAAAAAAGAA
ATTAAAGAAA
AAATAGTTTT
TGTTTTCCGA
AGATGTAAAA
GACTCTAGGG
GGATCGCCAA
CAAATACTAC
CTTTTATCTT
GCTCTTCCTG
CTCTCAGGTA
TTAATGCCGA
ATTGTTTCAT
CTTGTCTGTG
TAGAACACCA
CACACGAAAA
TCCTGTGATT
TTACATTTTA
CTTATCGTTA
ATCGAATGTA
TATCTATTTA
ATCTGCTTTT
CTTGTCTAAT
AAATATATAT
GTAAAGTACG
CTTTTTGTTG
AAATTTTTTA
AACCTTTGTT
TATTTTTTTT
TCTTCATTCC
GTAACTCTTC
TACCTTCTTT
ATTTACTTTC
TAAAATCCAA
ATACAAAACA
TAAAAATAAA
TAAACACAGA
GTAAATTCCC
AAATTATTCC
ATCATTAAAA
GATACGAGGC
GCGTGTAAGT
TACAGGCAAG
CGATCG