名称: | pEX18Tc-FlhA(PAO1) |
---|---|
别称: | PWU33336.1 |
基因长: | |
质粒宿主: | 大肠杆菌 |
质粒用途: | 基因编辑 |
片段类型: | |
片段物种: | |
原核抗性: | Tet |
筛选标记: | SacB |
荧光标记: | |
启动子: | |
复制子: | pBR322 |
拷贝数: | 低 |
感受态: | DH5a |
温度: | 37度 |
背景载体: | pEX18TC |
正向引物: | lac P |
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。
GACGAAAGGG
CCTCGTGATA
CGCCTATTTT
TATAGGTTAA
TGTCATGATA
ATAATGGTTT
CTTAGACGTC
AGGTGGCACT
TTTCGGGGAA
ATGTGCGCGG
AACCCCTATT
TGTTTATTTT
TCTAAATACA
TTCAAATATG
TATCCGCTCA
TGAGACAATA
ACCCTGATAA
ATGCTTCAAT
AATATTGAAA
AAGGAAGAGT
ATGAGTATTC
AACCGATTAG
CTTTAATGCG
GTAGTTTATC
ACAGTTAAAT
TGCTAACGCA
GTCAGGCACC
GTGTATGAAA
TCTAACAATG
CGCTCATCGT
CATCCTCGGC
ACCGTCACCC
TGGATGCTGT
AGGCATAGGC
TTGGTTATGC
CGGTACTGCC
GGGCCTCTTG
CGGGATATCG
TCCATTCCGA
CAGCATCGCC
AGTCACTATG
GCGTGCTGCT
AGCGCTATAT
GCGTTGATGC
AATTTCTATG
CGCACCCGTT
CTCGGAGCAC
TGTCCGACCG
CTTTGGCCGC
CGCCCAGTCC
TGCTCGCTTC
GCTACTTGGA
GCCACTATCG
ACTACGCGAT
CATGGCGACC
ACACCCGTCC
TGTGGATTCT
CTACGCCGGA
CGCATCGTGG
CCGGCATCAC
CGGCGCCACA
GGTGCGGTTG
CTGGCGCCTA
TATCGCCGAC
ATCACCGATG
GGGAAGATCG
GGCTCGCCAC
TTCGGGCTCA
TGAGCGCTTG
TTTCGGCGTG
GGTATGGTGG
CAGGCCCCGT
GGCCGGGGGA
CTGTTGGGCG
CCATCTCCTT
ACATGCACCA
TTCCTTGCGG
CGGCGGTGCT
CAACGGCCTC
AACCTACTAC
TGGGCTGCTT
CCTAATGCAG
GAGTCGCATA
AGGGAGAGCG
CCGACCGATG
CCCTTGAGAG
CCTTCAACCC
AGTCAGCTCC
TTCCGGTGGG
CGCGGGGCAT
GACTATCGTC
GCCGCACTTA
TGACTGTCTT
CTTTATCATG
CAACTCGTAG
GACAGGTGCC
GGCAGCGCTC
TGGGTCATTT
TCGGCGAGGA
CCGCTTTCGC
TGGAGCGCGA
CGATGATCGG
CCTGTCGCTT
GCGGTATTCG
GAATCTTGCA
CGCCCTCGCT
CAAGCCTTCG
TCACTGGTCC
CGCCACCAAA
CGTTTCGGCG
AGAAGCAGGC
CATTATCGCC
GGCATGGCGG
CCGACGCGCT
GGGCTACGTC
TTGCTGGCGT
TCGCGACGCG
AGGCTGGATG
GCCTTCCCCA
TTATGATTCT
TCTCGCTTCC
GGCGGCATCG
GGATGCCCGC
GTTGCAGGCC
ATGCTGTCCA
GGCAGGTAGA
TGACGACCAT
CAGGGACAGC
TTCAAGGATC
GCTCGCGGCT
CTTACCAGCC
TAACTTCGAT
CACTGGACCG
CTGATCGTCA
CGGCGATTTA
TGCCGCCTCG
GCGAGCACAT
GGAACGGGTT
GGCATGGATT
GTAGGCGCCG
CCCTATACCT
TGTCTGCCTC
CCCGCGTTGC
GTCGCGGTGC
ATGGAGCCGG
GCCACCTCGA
CCTGAATGGA
AGCCGGCGGC
ACCTCGCTAA
CGGATTCACC
ACTCCAAGAA
TTGGAGCCAA
TCAATTCTTG
CGGAGAACTG
TGAATGCGCA
AACCAACCCT
TGCATCGATT
GTCAGACCAA
GTTTACTCAT
ATATACTTTA
GATTGATTTA
AAACTTCATT
TTTAATTTAA
AAGGATCTAG
GTGAAGATCC
TTTTTGATAA
TCTCATGACC
AAAATCCCTT
AACGTGAGTT
TTCGTTCCAC
TGAGCGTCAG
ACCCCGTAGA
AAAGATCAAA
GGATCTTCTT
GAGATCCTTT
TTTTCTGCGC
GTAATCTGCT
GCTTGCAAAC
AAAAAAACCA
CCGCTACCAG
CGGTGGTTTG
TTTGCCGGAT
CAAGAGCTAC
CAACTCTTTT
TCCGAAGGTA
ACTGGCTTCA
GCAGAGCGCA
GATACCAAAT
ACTGTCCTTC
TAGTGTAGCC
GTAGTTAGGC
CACCACTTCA
AGAACTCTGT
AGCACCGCCT
ACATACCTCG
CTCTGCTAAT
CCTGTTACCA
GTGGCTGCTG
CCAGTGGCGA
TAAGTCGTGT
CTTACCGGGT
TGGACTCAAG
ACGATAGTTA
CCGGATAAGG
CGCAGCGGTC
GGGCTGAACG
GGGGGTTCGT
GCACACAGCC
CAGCTTGGAG
CGAACGACCT
ACACCGAACT
GAGATACCTA
CAGCGTGAGC
TATGAGAAAG
CGCCACGCTT
CCCGAAGGGA
GAAAGGCGGA
CAGGTATCCG
GTAAGCGGCA
GGGTCGGAAC
AGGAGAGCGC
ACGAGGGAGC
TTCCAGGGGG
AAACGCCTGG
TATCTTTATA
GTCCTGTCGG
GTTTCGCCAC
CTCTGACTTG
AGCGTCGATT
TTTGTGATGC
TCGTCAGGGG
GGCGGAGCCT
ATGGAAAAAC
GCCAGCAACG
CGGCCTTTTT
ACGGTTCCTG
GCCTTTTGCT
GGCCTTTTGC
TCACATGTTC
TTTCCTGCGT
TATCCCCTGA
TTCTGTGGAT
AACCGTATTA
CCGCCTTTGA
GTGAGCTGAT
ACCGCTCGCC
GCAGCCGAAC
GACCGAGCGC
AGCGAGTCAG
TGAGCGAGGA
AGCGGAAGAG
CGCCCAATAC
GCAAACCGCC
TCTCCCCGCG
CGTTGGCCGA
TTCATTAATG
CAGCTGGCAC
GACAGGTTTC
CCGACTGGAA
AGCGGGCAGT
GAGCGCAACG
CAATTAATGT
GAGTTAGCTC
ACTCATTAGG
CACCCCAGGC
TTTACACTTT
ATGCTTCCGG
CTCGTATGTT
GTGTGGAATT
GTGAGCGGAT
AACAATTTCA
CACAGGAAAC
AGCTATGACA
TGATTACGAA
TTCGTGGATC
GCACGCAACT
GATCAGCAAT
GTGCGCAGCA
ACCTGGCGGG
ACTGGGGCGC
GGCAGCCTCG
GCGTGCCGGT
CCTGCTGCTG
GCCATGCTGG
CGATGATGAC
CCTGCCGATC
CCGCCATTCC
TGCTGGACGT
GCTGTTCACC
TTCAACATCG
CCCTGTCCAT
CGTCGTCCTG
CTGGTCTGCG
TGTACGCCCT
GCGGCCGCTG
GACTTCGCCG
TGTTCCCGAC
CATCCTGCTG
GTGGCGACCC
TGTTGCGCCT
GGCGCTGAAC
GTTGCCTCGA
CCCGCGTGGT
GCTGCTCCAC
GGCCACGACG
GCCATGGCGC
CGCAGGCAAG
GTGATCCAGG
CCTTCGGCGA
AGTGGTGATC
GGCGGCAACT
ACGTGGTCGG
GGTGGTGGTG
TTCGCGATCC
TCATGATCAT
CAACTTCGTG
GTGGTGACCA
AGGGCGCCGG
GCGGATTTCC
GAAGTCAGCG
CGCGCTTCAC
CCTGGATGCC
ATGCCCGGCA
AGCAGATGGC
CATCGACGCC
GACCTCAACG
CCGGCCTGAT
CGAGCAGGAG
GAAGCCAAGA
AGCGCCGCAG
CGAGGTGGCC
CAGGAGGCCG
ACTTCTACGG
TTCGATGGAC
GGTGCCAGCA
AGTTCGTCCG
CGGCGACGCC
ATCGCCGGCC
TGCTGATTCT
CTTCATCAAC
CTGATCGGCG
GCGTCGCCAT
CGGCATGATC
CAGCATTCGA
TGTCCTTCGG
CGATGCCGGC
AAGGTCTACG
CCCTGCTGAC
CATCGGTGAC
GGCCTGGTGG
CACAACTGCC
GTCGCTGCTG
CTGTCGACCG
CGGCGGCGAT
CATGGTGACG
CGGGTCTCCA
GTTCCGAGGA
CATGGGCCAG
CAGGTCAACC
GGCAGATGTT
CGCCTCGCCC
AAGGCGCTGG
CGATTTCCGC
GGCGATCCTG
ATCGCCATGG
GCGCCGTGCC
GGGCATGCCG
CATGTTTCCT
TCATCGGCCT
CGGCGCCCTG
GCCGGCGCTG
CCGCCTACTG
GATCTGGCAC
CGGCAGAACA
AGGCCAAGCA
GGTCGCCGAG
CAGGAGGTGC
AGCGCCAGCA
GGAGCTGCTG
CCGGCGCAGC
GCGCCCAGGA
GGTCAAGGAA
CTGGGTTGGG
ACGACGTCAC
CCCGGTGGAC
ATGGTCGGCC
TGGAAGTCGG
CTACCGGCTG
ATCCCGCTGG
TCGACCGCAA
CCAGGGCGGG
CAACTGCTGG
CGCGGATCAA
GGGCGTACGC
AAGAAGCTGT
CCCAGGAGCT
GGGCTTCCTG
ATGCCCTCGG
TGCACATCCG
CGACAACCTC
GACCTGCTGC
CCAACGCCTA
CCGTCTGACC
CTGATGGGGG
TCAGCGTGGC
CGAAGCGGAG
ATCTATCCCG
ATCGCGAGCT
GGCGATCAAC
CCCGGCCAGG
TATTCGGCAG
CCTCAACGGC
ATCGCCGGCA
AGGACCCGGC
CTTCGGCCTG
GAGGCGGTAT
GGATCGAGCC
CGGCCAGCGC
GACCAGGCGC
AGTCGCTGGG
CTACACCGTG
GTGGATGCGA
GCACAGTGGT
CGCCACTCAC
CTGAATCAGG
TGCTGCACAA
GCACGCCCAC
GAGCTGCTGG
GCCATGAAGA
AGTGCAACAG
CTCCTGCAAT
TGCTCGCGCG
CAGCTCGCCG
AAGCTGGCGG
AAGAGCTGGT
GCCGGGGCTG
ATTTCCCTGT
CCACCCTGCT
CAAGGTCCTG
CAGGCGCTGC
TCCAGGAGCA
GGTGCCGGTG
CGCGACATCC
GTACCATCGC
CGAGGCCATC
GCCAACGTCG
CTCCACGGAG
TCAAGATCCC
GGCGCGATGG
TCGCCGCGGT
TCGCGTCGCG
TTGTCCCGCG
CAATCGTGCA
AAGCATTGTC
GGGCTTGAGC
CGGAGCTGCC
TGTGATCACG
CTCGAGCCGA
GGTTGGAACA
GATTTTGCTG
AACAGCATGC
AGAAGGCCGG
ACAAGGCTCC
GAGGACGGCA
TGTTGCTGGA
GCCGGGGATG
GCCGAGAAGC
TGCAACGCTC
GCTGATCGAG
AGCGCGCAAC
GCCAGGAGAT
GCTCGGTAAG
CCGGCGATCC
TGCTGGTTGC
CGGACCGATC
CGCTCGATGA
TGTCGCGCTT
CGCGCGGATG
GCGGTGCCCA
GCATGCATGT
GCTGGCCTAC
CAGGAAATCC
CGGACAACAA
GCAAGTCACC
ATCGTCGCGA
CGGTGGGACA
GAACTGAGGA
TCCTCTAGAG
TCGACCTGCA
GGCATGCAAG
CTTGGCACTG
GCCGTCGTTT
TACAACGTCG
TGACTGGGAA
AACCCTGGCG
TTACCCAACT
TAATCGCCTT
GCAGCACATC
CCCCTTTCGC
CAGCTGGCGT
AATAGCGAAG
AGGCCCGCAC
CGATCGCCCT
TCCCAACAGT
TGCGCAGCCT
GAATGGCGAA
TGGCGAGCTT
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
AACTCTTTTT
GTTTATTTTT
CTAAATACAT
TCAAATATGC
GCCTGATGCG
GTATTTTCTC
CTTACGCATA
TCGACATCCG
CCCTCACCGC
CAGGAACGCA
ACCGCAGCCT
CATCACGCCG
GCGCTTCTTG
GCCGCGCGGG
ATTCAACCCA
CTCGGCCAGC
TCGTCGGTGT
AGCTCTTTGG
CATCGTCTCT
CGCCTGTCCC
CTCAGTTCAG
TAATTTCCTG
CATTTGCCTG
TTTCCAGTCG
GTAGATATTC
CACAAAACAG
CAGGGAAGCA
GCGCTTTTCC
GCTGCATAAC
CCTGCTTCGG
GGTCATTATA
GCGATTTTTT
CGGTATATCC
ATCCTTTTTC
GCACGATATA
CAGGATTTTG
CCAAAGGGTT
CGTGTAGACT
TTCCTTGGTG
TATCCAACGG
CGTCAGCCGG
GCAGGATAGG
TGAAGTAGGC
CCACCCGCGA
GCGGGTGTTC
CTTCTTCACT
GTCCCTTATT
CGCACCTGGC
GGTGCTCAAC
GGGAATCCTG
CTCTGCGAGG
CTGGCCGGCT
ACCGCCGGCG
TAACAGATGA
GGGCAAGCGG
ATGGCTGATG
AAACCAAGCC
AACCAGGAAG
GGCAGCCCAC
CTATCAAGGT
GTACTGCCTT
CCAGACGAAC
GAAGAGCGAT
TGAGGAAAAG
GCGGCGGCGG
CCGGCATGAG
CCTGTCGGCC
TACCTGCTGG
CCGTCGGCCA
GGGCTACAAA
ATCACGGGCG
TCGTGGACTA
TGAGCACGTC
CGCGAGCTGG
CCCGCATCAA
TGGCGACCTG
GGCCGCCTGG
GCGGCCTGCT
GAAACTCTGG
CTCACCGACG
ACCCGCGCAC
GGCGCGGTTC
GGTGATGCCA
CGATCCTCGC
CCTGCTGGCG
AAGATCGACT
CTAGCTAGAG
GATCGATCCT
TTTTAACCCA
TCACATATAC
CTGCCGTTCA
CTATTATTTA
GTGAAATGAG
ATATTATGAT
ATTTTCTGAA
TTGTGATTAA
AAAGGCAACT
TTATGCCCAT
GCAACAGAAA
CTATAAAAAA
TACAGAGAAT
GAAAAGAAAC
AGATAGATTT
TTTAGTTCTT
TAGGCCCGTA
GTCTGCAAAT
CCTTTTATGA
TTTTCTATCA
AACAAAAGAG
GAAAATAGAC
CAGTTGCAAT
CCAAACGAGA
GTCTAATAGA
ATGAGGTCGA
AAAGTAAATC
GCGCGGGTTT
GTTACTGATA
AAGCAGGCAA
GACCTAAAAT
GTGTAAAGGG
CAAAGTGTAT
ACTTTGGCGT
CACCCCTTAC
ATATTTTAGG
TCTTTTTTTA
TTGTGCGTAA
CTAACTTGCC
ATCTTCAAAC
AGGAGGGCTG
GAAGAAGCAG
ACCGCTAACA
CAGTACATAA
AAAAGGAGAC
ATGAACGATG
AACATCAAAA
AGTTTGCAAA
ACAAGCAACA
GTATTAACCT
TTACTACCGC
ACTGCTGGCA
GGAGGCGCAA
CTCAAGCGTT
TGCGAAAGAA
ACGAACCAAA
AGCCATATAA
GGAAACATAC
GGCATTTCCC
ATATTACACG
CCATGATATG
CTGCAAATCC
CTGAACAGCA
AAAAAATGAA
AAATATCAAG
TTCCTGAGTT
CGATTCGTCC
ACAATTAAAA
ATATCTCTTC
TGCAAAAGGC
CTGGACGTTT
GGGACAGCTG
GCCATTACAA
AACGCTGACG
GCACTGTCGC
AAACTATCAC
GGCTACCACA
TCGTCTTTGC
ATTAGCCGGA
GATCCTAAAA
ATGCGGATGA
CACATCGATT
TACATGTTCT
ATCAAAAAGT
CGGCGAAACT
TCTATTGACA
GCTGGAAAAA
CGCTGGCCGC
GTCTTTAAAG
ACAGCGACAA
ATTCGATGCA
AATGATTCTA
TCCTAAAAGA
CCAAACACAA
GAATGGTCAG
GTTCAGCCAC
ATTTACATCT
GACGGAAAAA
TCCGTTTATT
CTACACTGAT
TTCTCCGGTA
AACATTACGG
CAAACAAACA
CTGACAACTG
CACAAGTTAA
CGTATCAGCA
TCAGACAGCT
CTTTGAACAT
CAACGGTGTA
GAGGATTATA
AATCAATCTT
TGACGGTGAC
GGAAAAACGT
ATCAAAATGT
ACAGCAGTTC
ATCGATGAAG
GCAACTACAG
CTCAGGCGAC
AACCATACGC
TGAGAGATCC
TCACTACGTA
GAAGATAAAG
GCCACAAATA
CTTAGTATTT
GAAGCAAACA
CTGGAACTGA
AGATGGCTAC
CAAGGCGAAG
AATCTTTATT
TAACAAAGCA
TACTATGGCA
AAAGCACATC
ATTCTTCCGT
CAAGAAAGTC
AAAAACTTCT
GCAAAGCGAT
AAAAAACGCA
CGGCTGAGTT
AGCAAACGGC
GCTCTCGGTA
TGATTGAGCT
AAACGATGAT
TACACACTGA
AAAAAGTGAT
GAAACCGCTG
ATTGCATCTA
ACACAGTAAC
AGATGAAATT
GAACGCGCGA
ACGTCTTTAA
AATGAACGGC
AAATGGTATC
TGTTCACTGA
CTCCCGCGGA
TCAAAAATGA
CGATTGACGG
CATTACGTCT
AACGATATTT
ACATGCTTGG
TTATGTTTCT
AATTCTTTAA
CTGGCCCATA
CAAGCCGCTG
AACAAAACTG
GCCTTGTGTT
AAAAATGGAT
CTTGATCCTA
ACGATGTAAC
CTTTACTTAC
TCACACTTCG
CTGTACCTCA
AGCGAAAGGA
AACAATGTCG
TGATTACAAG
CTATATGACA
AACAGAGGAT
TCTACGCAGA
CAAACAATCA
ACGTTTGCGC
CTAGCTTCCT
GCTGAACATC
AAAGGCAAGA
AAACATCTGT
TGTCAAAGAC
AGCATCCTTG
AACAAGGACA
ATTAACAGTT
AACAAATAAA
AACGCAAAAG
AAAATGCCGA
TTATGGTGCA
CTCTCAGTAC
AATCTGCTCT
GATGCCGCAT
AGTTAAGCCA
GCCCCGACAC
CCGCCAACAC
CCGCTGACGC
GCCCTGACGG
GCTTGTCTGC
TCCCGGCATC
CGCTTACAGA
CAAGCTGTGA
CCGTCTCCGG
GAGCTGCATG
TGTCAGAGGT
TTTCACCGTC
ATCACCGAAA
CGCGCGA