名称: | pYD1-NRK1(opt-yeast) |
---|---|
别称: | XM_022820921.1 |
基因长: | 714bp |
质粒宿主: | 酵母菌 |
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: | TRP1 |
荧光标记: | |
启动子: | GAL1 |
复制子: | CEN/ARS |
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。
ACGGATTAGA
AGCCGCCGAG
CGGGTGACAG
CCCTCCGAAG
GAAGACTCTC
CTCCGTGCGT
CCTCGTCTTC
ACCGGTCGCG
TTCCTGAAAC
GCAGATGTGC
CTCGCGCCGC
ACTGCTCCGA
ACAATAAAGA
TTCTACAATA
CTAGCTTTTA
TGGTTATGAA
GAGGAAAAAT
TGGCAGTAAC
CTGGCCCCAC
AAACCTTCAA
ATGAACGAAT
CAAATTAACA
ACCATAGGAT
GATAATGCGA
TTAGTTTTTT
AGCCTTATTT
CTGGGGTAAT
TAATCAGCGA
AGCGATGATT
TTTGATCTAT
TAACAGATAT
ATAAATGCAA
AAACTGCATA
ACCACTTTAA
CTAATACTTT
CAACATTTTC
GGTTTGTATT
ACTTCTTATT
CAAATGTAAT
AAAAGTATCA
ACAAAAAATT
GTTAATATAC
CTCTATACTT
TAACGTCAAG
GAGAAAAAAC
CCCGGATCGG
ACTACTAGCA
GCTGTAATAC
GACTCACTAT
AGGGAATATT
AAGCTAATTC
CCTACTTCAT
ACATTTTCAA
TTAAGATGCA
GTTACTTCGC
TGTTTTTCAA
TATTTTCTGT
TATTGCTTCA
GTTTTAGCAC
AGGAACTGAC
AACTATATGC
GAGCAAATCC
CCTCACCAAC
TTTAGAATCG
ACGCCGTACT
CTTTGTCAAC
GACTACTATT
TTGGCCAACG
GGAAGGCAAT
GCAAGGAGTT
TTTGAATATT
ACAAATCAGT
AACGTTTGTC
AGTAATTGCG
GTTCTCACCC
CTCAACAACT
AGCAAAGGCA
GCCCCATAAA
CACACAGTAT
GTTTTTAAGC
TTCTGCAGGC
TAGTGGTGGT
GGTGGTTCTG
GTGGTGGTGG
TTCTGGTGGT
GGTGGTTCTG
CTAGCATGAC
TGGTGGACAG
CAAATGGGTC
GGGATCTGTA
CGACGATGAC
GATAAGGTAC
CAGGATCCAG
TGTGGTGGAA
TTCGACTACA
AGGACGACGA
CGACAAGATG
ACAACTACTA
AGGTTAAGCT
AATTGCTATA
TCAGGTTGTT
CATCTTCCGG
TAAAACAACC
TTGGCTAAAT
TCCTTGCCAA
CGCTATTCCA
GGTTGTATAC
TTATTCATGA
AGATGACTTT
TACAAACCAG
ATTCAGAAAT
TCCAATCAAT
GAAAAATATG
GTGTTGCCGA
CTGGGATTGT
CCTGAGGCTC
TGGACTTAGA
TGCCTTCAAA
AGAGAACTTG
ATTTGATTAA
GACTACTGGC
TCCATTAAGA
CGAAGTTGAT
TCATAACGAG
AATGTTGACG
ACATCGGTAA
GTTTAATATA
AAGCAAGAGG
ATTGGGACGC
TCTTCGTGCC
AAGTTATCCT
CAGTTATTGA
AAGTGATTTA
AAAGTCGTAC
TAGTAGACGG
ATTTATGATC
TTTAATGACG
AAGAATTGAT
GAGAAAATTT
GATATAAGGA
TATTTGTAAG
AGCACCATAC
GAAGTTTTAT
CAAGAAGACG
TCATGCAAGG
GCCGGCTACA
AAACACTAGA
ATCATTCTGG
GTTGACCCAC
CATATTATTT
CGATGAGTTC
GTCTATAGAG
CATATAGAGA
AGAACATAAA
CACCTTTTCG
TGAATGAAGA
CGTAGAAGGG
TCATTGAGAA
GTGATGCGGG
TTTATTTGAA
TTAATCAATG
ACGACGAAAC
CGAAATAACT
ACTGCTCTAA
ACACCATTGC
TGACTACATT
GTTTCACACT
TGGATGCTAA
TTAACTCGAG
TCTAGAGGGC
CCTTCGAAGG
TAAGCCTATC
CCTAACCCTC
TCCTCGGTCT
CGATTCTACG
CGTACCGGTC
ATCATCACCA
TCACCATTGA
GTTTAAACCC
GCTGATCTGA
TAACAACAGT
GTAGATGTAA
CAAAATCGAC
TTTGTTCCCA
CTGTACTTTT
AGCTCGTACA
AAATACAATA
TACTTTTCAT
TTCTCCGTAA
ACAACATGTT
TTCCCATGTA
ATATCCTTTT
CTATTTTTCG
TTCCGTTACC
AACTTTACAC
ATACTTTATA
TAGCTATTCA
CTTCTATACA
CTAAAAAACT
AAGACAATTT
TAATTTTGCT
GCCTGCCATA
TTTCAATTTG
TTATAAATTC
CTATAATTTA
TCCTATTAGT
AGCTAAAAAA
AGATGAATGT
GAATCGAATC
CTAAGAGAAT
TGGGCAAGTG
CACAAACAAT
ACTTAAATAA
ATACTACTCA
GTAATAACCT
ATTTCTTAGC
ATTTTTGACG
AAATTTGCTA
TTTTGTTAGA
GTCTTTTACA
CCATTTGTCT
CCACACCTCC
GCTTACATCA
ACACCAATAA
CGCCATTTAA
TCTAAGCGCA
TCACCAACAT
TTTCTGGCGT
CAGTCCACCA
GCTAACATAA
AATGTAAGCT
CTCGGGGCTC
TCTTGCCTTC
CAACCCAGTC
AGAAATCGAG
TTCCAATCCA
AAAGTTCACC
TGTCCCACCT
GCTTCTGAAT
CAAACAAGGG
AATAAACGAA
TGAGGTTTCT
GTGAAGCTGC
ACTGAGTAGT
ATGTTGCAGT
CTTTTGGAAA
TACGAGTCTT
TTAATAACTG
GCAAACCGAG
GAACTCTTGG
TATTCTTGCC
ACGACTCATC
TCCGTGCAGT
TGGACGATAT
CAATGCCGTA
ATCATTGACC
AGAGCCAAAA
CATCCTCCTT
AGGTTGATTA
CGAAACACGC
CAACCAAGTA
TTTCGGAGTG
CCTGAACTAT
TTTTATATGC
TTTTACAAGA
CTTGAAATTT
TCCTTGCAAT
AACCGGGTCA
ATTGTTCTCT
TTCTATTGGG
CACACATATA
ATACCCAGCA
AGTCAGCATC
GGAATCTAGA
GCACATTCTG
CGGCCTCTGT
GCTCTGCAAG
CCGCAAACTT
TCACCAATGG
ACCAGAACTA
CCTGTGAAAT
TAATAACAGA
CATACTCCAA
GCTGCCTTTG
TGTGCTTAAT
CACGTATACT
CACGTGCTCA
ATAGTCACCA
ATGCCCTCCC
TCTTGGCCCT
CTCCTTTTCT
TTTTTCGACC
GAATTTCTTG
AAGACGAAAG
GGCCTCGTGA
TACGCCTATT
TTTATAGGTT
AATGTCATGA
TAATAATGGT
TTCTTAGGAC
GGATCGCTTG
CCTGTAACTT
ACACGCGCCT
CGTATCTTTT
AATGATGGAA
TAATTTGGGA
ATTTACTCTG
TGTTTATTTA
TTTTTATGTT
TTGTATTTGG
ATTTTAGAAA
GTAAATAAAG
AAGGTAGAAG
AGTTACGGAA
TGAAGAAAAA
AAAATAAACA
AAGGTTTAAA
AAATTTCAAC
AAAAAGCGTA
CTTTACATAT
ATATTTATTA
GACAAGAAAA
GCAGATTAAA
TAGATATACA
TTCGATTAAC
GATAAGTAAA
ATGTAAAATC
ACAGGATTTT
CGTGTGTGGT
CTTCTACACA
GACAAGATGA
AACAATTCGG
CATTAATACC
TGAGAGCAGG
AAGAGCAAGA
TAAAAGGTAG
TATTTGTTGG
CGATCCCCCT
AGAGTCTTTT
ACATCTTCGG
AAAACAAAAA
CTATTTTTTC
TTTAATTTCT
TTTTTTACTT
TCTATTTTTA
ATTTATATAT
TTATATTAAA
AAATTTAAAT
TATAATTATT
TTTATAGCAC
GTGATGAAAA
GGACCCAGGT
GGCACTTTTC
GGGGAAATGT
GCGCGGAACC
CCTATTTGTT
TATTTTTCTA
AATACATTCA
AATATGTATC
CGCTCATGAG
ACAATAACCC
TGATAAATGC
TTCAATAATA
TTGAAAAAGG
AAGAGTATGA
GTATTCAACA
TTTCCGTGTC
GCCCTTATTC
CCTTTTTTGC
GGCATTTTGC
CTTCCTGTTT
TTGCTCACCC
AGAAACGCTG
GTGAAAGTAA
AAGATGCTGA
AGATCAGTTG
GGTGCACGAG
TGGGTTACAT
CGAACTGGAT
CTCAACAGCG
GTAAGATCCT
TGAGAGTTTT
CGCCCCGAAG
AACGTTTTCC
AATGATGAGC
ACTTTTAAAG
TTCTGCTATG
TGGCGCGGTA
TTATCCCGTG
TTGACGCCGG
GCAAGAGCAA
CTCGGTCGCC
GCATACACTA
TTCTCAGAAT
GACTTGGTTG
AGTACTCACC
AGTCACAGAA
AAGCATCTTA
CGGATGGCAT
GACAGTAAGA
GAATTATGCA
GTGCTGCCAT
AACCATGAGT
GATAACACTG
CGGCCAACTT
ACTTCTGACA
ACGATCGGAG
GACCGAAGGA
GCTAACCGCT
TTTTTGCACA
ACATGGGGGA
TCATGTAACT
CGCCTTGATC
GTTGGGAACC
GGAGCTGAAT
GAAGCCATAC
CAAACGACGA
GCGTGACACC
ACGATGCCTG
TAGCAATGGC
AACAACGTTG
CGCAAACTAT
TAACTGGCGA
ACTACTTACT
CTAGCTTCCC
GGCAACAATT
AATAGACTGG
ATGGAGGCGG
ATAAAGTTGC
AGGACCACTT
CTGCGCTCGG
CCCTTCCGGC
TGGCTGGTTT
ATTGCTGATA
AATCTGGAGC
CGGTGAGCGT
GGGTCTCGCG
GTATCATTGC
AGCACTGGGG
CCAGATGGTA
AGCCCTCCCG
TATCGTAGTT
ATCTACACGA
CGGGCAGTCA
GGCAACTATG
GATGAACGAA
ATAGACAGAT
CGCTGAGATA
GGTGCCTCAC
TGATTAAGCA
TTGGTAACTG
TCAGACCAAG
TTTACTCATA
TATACTTTAG
ATTGATTTAA
AACTTCATTT
TTAATTTAAA
AGGATCTAGG
TGAAGATCCT
TTTTGATAAT
CTCATGACCA
AAATCCCTTA
ACGTGAGTTT
TCGTTCCACT
GAGCGTCAGA
CCCCGTAGAA
AAGATCAAAG
GATCTTCTTG
AGATCCTTTT
TTTCTGCGCG
TAATCTGCTG
CTTGCAAACA
AAAAAACCAC
CGCTACCAGC
GGTGGTTTGT
TTGCCGGATC
AAGAGCTACC
AACTCTTTTT
CCGAAGGTAA
CTGGCTTCAG
CAGAGCGCAG
ATACCAAATA
CTGTCCTTCT
AGTGTAGCCG
TAGTTAGGCC
ACCACTTCAA
GAACTCTGTA
GCACCGCCTA
CATACCTCGC
TCTGCTAATC
CTGTTACCAG
TGGCTGCTGC
CAGTGGCGAT
AAGTCGTGTC
TTACCGGGTT
GGACTCAAGA
CGATAGTTAC
CGGATAAGGC
GCAGCGGTCG
GGCTGAACGG
GGGGTTCGTG
CACACAGCCC
AGCTTGGAGC
GAACGACCTA
CACCGAACTG
AGATACCTAC
AGCGTGAGCA
TTGAGAAAGC
GCCACGCTTC
CCGAAGGGAG
AAAGGCGGAC
AGGTATCCGG
TAAGCGGCAG
GGTCGGAACA
GGAGAGCGCA
CGAGGGAGCT
TCCAGGGGGG
AACGCCTGGT
ATCTTTATAG
TCCTGTCGGG
TTTCGCCACC
TCTGACTTGA
GCGTCGATTT
TTGTGATGCT
CGTCAGGGGG
GCCGAGCCTA
TGGAAAAACG
CCAGCAACGC
GGCCTTTTTA
CGGTTCCTGG
CCTTTTGCTG
GCCTTTTGCT
CACATGTTCT
TTCCTGCGTT
ATCCCCTGAT
TCTGTGGATA
ACCGTATTAC
CGCCTTTGAG
TGAGCTGATA
CCGCTCGCCG
CAGCCGAACG
ACCGAGCGCA
GCGAGTCAGT
GAGCGAGGAA
GCGGAAGAGC
GCCCAATACG
CAAACCGCCT
CTCCCCGCGC
GTTGGCCGAT
TCATTAATGC
AGCTGGCACG
ACAGGTTTCC
CGACTGGAAA
GCGGGCAGTG
AGCGCAACGC
AATTAATGTG
AGTTACCTCA
CTCATTAGGC
ACCCCAGGCT
TTACACTTTA
TGCTTCCGGC
TCCTATGTTG
TGTGGAATTG
TGAGCGGATA
ACAATTTCAC
ACAGGAAACA
GCTATGACCA
TGATTACGCC
AAGCTCGGAA
TTAACCCTCA
CTAAAGGGAA
CAAAAGCTGG
CTAGT