名称: | pSIN-EF2-Steep1(mouse)-HA-Puro |
---|---|
别称: | NM_029951.1,Steep1/C330007P06Rik/STEEP |
基因长: | 666bp |
质粒宿主: | 哺乳细胞,慢病毒 |
质粒用途: | 蛋白表达 |
片段类型: | ORF |
片段物种: | 小鼠 |
原核抗性: | Amp |
筛选标记: | Puro |
荧光标记: | |
启动子: | EF1a |
复制子: | pUC |
拷贝数: | |
感受态: | Stbl3 |
温度: | 37度 |
背景载体: | pSIN-EF2-puro |
正向引物: | PEF-F |
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。
CGGACCGCCA
CTGCCAATTA
CCTGTGGTTT
CATTTACTCT
AAACCTGTGA
TTCCTCTGAA
TTATTTTCAT
TTTAAAGAAA
TTGTATTTGT
TAAATATGTA
CTACAAACTT
AGTAGTTGGA
AGGGCTAATT
CACTCCCAAA
GAAGACAAGA
TATCCTTGAT
CTGTGGATCT
ACCACACACA
AGGCTACTTC
CCTGATTAGC
AGAACTACAC
ACCAGGGCCA
GGGGTCAGAT
ATCCACTGAC
CTTTGGATGG
TGCTACAAGC
TAGTACCAGT
TGAGCCAGAT
AAGGTAGAAG
AGGCCAATAA
AGGAGAGAAC
ACCAGCTTGT
TACACCCTGT
GAGCCTGCAT
GGGATGGATG
ACCCGGAGAG
AGAAGTGTTA
GAGTGGAGGT
TTGACAGCCG
CCTAGCATTT
CATCACGTGG
CCCGAGAGCT
GCATCCGGAG
TACTTCAAGA
ACTGCTGATA
TCGAGCTTGC
TACAAGGGAC
TTTCCGCTGG
GGACTTTCCA
GGGAGGCGTG
GCCTGGGCGG
GACTGGGGAG
TGGCGAGCCC
TCAGATCCTG
CATATAAGCA
GCTGCTTTTT
GCCTGTACTG
GGTCTCTCTG
GTTAGACCAG
ATCTGAGCCT
GGGAGCTCTC
TGGCTAACTA
GGGAACCCAC
TGCTTAAGCC
TCAATAAAGC
TTGCCTTGAG
TGCTTCAAGT
AGTGTGTGCC
CGTCTGTTGT
GTGACTCTGG
TAACTAGAGA
TCCCTCAGAC
CCTTTTAGTC
AGTGTGGAAA
ATCTCTAGCA
GTGGCGCCCG
AACAGGGACT
TGAAAGCGAA
AGGGAAACCA
GAGGAGCTCT
CTCGACGCAG
GACTCGGCTT
GCTGAAGCGC
GCACGGCAAG
AGGCGAGGGG
CGGCGACTGG
TGAGTACGCC
AAAAATTTTG
ACTAGCGGAG
GCTAGAAGGA
GAGAGATGGG
TGCGAGAGCG
TCAGTATTAA
GCGGGGGAGA
ATTAGATCGC
GATGGGAAAA
AATTCGGTTA
AGGCCAGGGG
GAAAGAAAAA
ATATAAATTA
AAACATATAG
TATGGGCAAG
CAGGGAGCTA
GAACGATTCG
CAGTTAATCC
TGGCCTGTTA
GAAACATCAG
AAGGCTGTAG
ACAAATACTG
GGACAGCTAC
AACCATCCCT
TCAGACAGGA
TCAGAAGAAC
TTAGATCATT
ATATAATACA
GTAGCAACCC
TCTATTGTGT
GCATCAAAGG
ATAGAGATAA
AAGACACCAA
GGAAGCTTTA
GACAAGATAG
AGGAAGAGCA
AAACAAAAGT
AAGACCACCG
CACAGCAAGC
GGCCGCTGAT
CTTCAGACCT
GGAGGAGGAG
ATATGAGGGA
CAATTGGAGA
AGTGAATTAT
ATAAATATAA
AGTAGTAAAA
ATTGAACCAT
TAGGAGTAGC
ACCCACCAAG
GCAAAGAGAA
GAGTGGTGCA
GAGAGAAAAA
AGAGCAGTGG
GAATAGGAGC
TTTGTTCCTT
GGGTTCTTGG
GAGCAGCAGG
AAGCACTATG
GGCGCAGCGT
CAATGACGCT
GACGGTACAG
GCCAGACAAT
TATTGTCTGG
TATAGTGCAG
CAGCAGAACA
ATTTGCTGAG
GGCTATTGAG
GCGCAACAGC
ATCTGTTGCA
ACTCACAGTC
TGGGGCATCA
AGCAGCTCCA
GGCAAGAATC
CTGGCTGTGG
AAAGATACCT
AAAGGATCAA
CAGCTCCTGG
GGATTTGGGG
TTGCTCTGGA
AAACTCATTT
GCACCACTGC
TGTGCCTTGG
AATGCTAGTT
GGAGTAATAA
ATCTCTGGAA
CAGATTTGGA
ATCACACGAC
CTGGATGGAG
TGGGACAGAG
AAATTAACAA
TTACACAAGC
TTAATACACT
CCTTAATTGA
AGAATCGCAA
AACCAGCAAG
AAAAGAATGA
ACAAGAATTA
TTGGAATTAG
ATAAATGGGC
AAGTTTGTGG
AATTGGTTTA
ACATAACAAA
TTGGCTGTGG
TATATAAAAT
TATTCATAAT
GATAGTAGGA
GGCTTGGTAG
GTTTAAGAAT
AGTTTTTGCT
GTACTTTCTA
TAGTGAATAG
AGTTAGGCAG
GGATATTCAC
CATTATCGTT
TCAGACCCAC
CTCCCAACCC
CGAGGGGACC
CGACAGGCCC
GAAGGAATAG
AAGAAGAAGG
TGGAGAGAGA
GACAGAGACA
GATCCATTCG
ATTAGTGAAC
GGATCTCGAC
GGTATCGCCA
CAAATGGCAG
TATTCATCCA
CAATTTTAAA
AGAAAGGGGG
GGATTGGGGG
GTACAGTGCA
GGGGAAAGAA
TAGTAGACAT
AATAGCAACA
GACATACAAA
CTAAAGAATT
ACAAAAACAA
ATTACAAAAA
TTCAAAATTT
TCGGGTTTAT
TACAGGGACA
GCAGAGATCC
ACTTTGGATC
GATAAGCTTT
GCAAAGATGG
ATAAAGTTTT
AAACAGAGAG
GAATCTTTGC
AGCTAATGGA
CCTTCTAGGT
CTTGAAAGGA
GTGGGAATTG
GCTCCGGTGC
CCGTCAGTGG
GCAGAGCGCA
CATCGCCCAC
AGTCCCCGAG
AAGTTGGGGG
GAGGGGTCGG
CAATTGAACC
GGTGCCTAGA
GAAGGTGGCG
CGGGGTAAAC
TGGGAAAGTG
ATGTCGTGTA
CTGGCTCCGC
CTTTTTCCCG
AGGGTGGGGG
AGAACCGTAT
ATAAGTGCAG
TAGTCGCCGT
GAACGTTCTT
TTTCGCAACG
GGTTTGCCGC
CAGAACACAG
GTAAGTGCCG
TGTGTGGTTC
CCGCGGGCCT
GGCCTCTTTA
CGGGTTATGG
CCCTTGCGTG
CCTTGAATTA
CTTCCACCTG
GCTGCAGTAC
GTGATTCTTG
ATCCCGAGCT
TCGGGTTGGA
AGTGGGTGGG
AGAGTTCGAG
GCCTTGCGCT
TAAGGAGCCC
CTTCGCCTCG
TGCTTGAGTT
GAGGCCTGGC
CTGGGCGCTG
GGGCCGCCGC
GTGCGAATCT
GGTGGCACCT
TCGCGCCTGT
CTCGCTGCTT
TCGATAAGTC
TCTAGCCATT
TAAAATTTTT
GATGACCTGC
TGCGACGCTT
TTTTTCTGGC
AAGATAGTCT
TGTAAATGCG
GGCCAAGATC
TGCACACTGG
TATTTCGGTT
TTTGGGGCCG
CGGGCGGCGA
CGGGGCCCGT
GCGTCCCAGC
GCACATGTTC
GGCGAGGCGG
GGCCTGCGAG
CGCGGCCACC
GAGAATCGGA
CGGGGGTAGT
CTCAAGCTGG
CCGGCCTGCT
CTGGTGCCTG
GCCTCGCGCC
GCCGTGTATC
GCCCCGCCCT
GGGCGGCAAG
GCTGGCCCGG
TCGGCACCAG
TTGCGTGAGC
GGAAAGATGG
CCGCTTCCCG
GCCCTGCTGC
AGGGAGCTCA
AAATGGAGGA
CGCGGCGCTC
GGGAGAGCGG
GCGGGTGAGT
CACCCACACA
AAGGAAAAGG
GCCTTTCCGT
CCTCAGCCGT
CGCTTCATGT
GACTCCACGG
AGTACCGGGC
GCCGTCCAGG
CACCTCGATT
AGTTCTCGAG
CTTTTGGAGT
ACGTCGTCTT
TAGGTTGGGG
GGAGGGGTTT
TATGCGATGG
AGTTTCCCCA
CACTGAGTGG
GTGGAGACTG
AAGTTAGGCC
AGCTTGGCAC
TTGATGTAAT
TCTCCTTGGA
ATTTGCCCTT
TTTGAGTTTG
GATCTTGGTT
CATTCTCAAG
CCTCAGACAG
TGGTTCAAAG
TTTTTTTCTT
CCATTTCAGG
TGTCGTGAGG
AATTGGGATC
CGCCACCATG
CCGAAAGTCG
TGTCGCGGTC
GGTGGTCTGC
TCAGACACCC
GGGACCGCGA
GGAATATGAT
GATGGCGAGA
AGCCCCTCCA
TGTCTACTAC
TGTTTATGTG
GCCAGATGGT
CCTGGTGCTG
GATTGCCAGT
TAGAGAAATT
GCCCATGAGG
CCCCGGGACC
GGTCTCGCGT
GATCGATGCT
GCCAAACATG
CTCACAAGTT
TTGTAACACA
GAAGATGAGG
AGACTACTTA
TCTTCGGAGG
CCTGAGGGCA
TTGAACGTCA
GTACAGGAAG
AAGTGTGCAA
AGTGTGGACT
GCCGCTCTTC
TACCAATCCC
AGCCGAAGAA
TGCTCCTGTG
ACCTTCATTG
TGGATGGAGC
AGTAGTGAAA
TTTGGCCAAG
GCTTTGGGAA
AACCAATATA
TATACTCAGA
AACAAGAGCC
TCCTAAGAAG
GTTATGATGA
CCAAAAGGAC
TAAAGACATG
GGCAAGTTCA
GCTCTGTTAC
TGTGTCTACC
ATTGATGAAG
AAGAAGAGGA
GATAGAGGCT
AGGGAAGTTG
CTGACTCTTA
CGCGCAGAAT
GCCAAAGTGA
TTGAAAAGCA
GCTGGAGCGC
AAAGGCATGA
GCAAGAGGAG
GCTGCAGGAG
TTGGCTGAAT
TGGAAGCCAA
GAAAGCAAAA
ATGAAGGGGA
CCCTGATCGA
CAATCAGTTC
AAATACCCAT
ACGACGTACC
AGATTACGCT
TGAGAATTCT
TCGAACATAT
GAAGCTTGCT
AGCACTAGTG
ATCCGCATGC
ATCTAGGGCG
GCCAATTCCG
CCCCTCTCCC
CCCCCCCCCT
CTCCCTCCCC
CCCCCCTAAC
GTTACTGGCC
GAAGCCGCTT
GGAATAAGGC
CGGTGTGCGT
TTGTCTATAT
GTTATTTTCC
ACCATATTGC
CGTCTTTTGG
CAATGTGAGG
GCCCGGAAAC
CTGGCCCTGT
CTTCTTGACG
AGCATTCCTA
GGGGTCTTTC
CCCTCTCGCC
AAAGGAATGC
AAGGTCTGTT
GAATGTCGTG
AAGGAAGCAG
TTCCTCTGGA
AGCTTCTTGA
AGACAAACAA
CGTCTGTAGC
GACCCTTTGC
AGGCAGCGGA
ACCCCCCACC
TGGCGACAGG
TGCCTCTGCG
GCCAAAAGCC
ACGTGTATAA
GATACACCTG
CAAAGGCGGC
ACAACCCCAG
TGCCACGTTG
TGAGTTGGAT
AGTTGTGGAA
AGAGTCAAAT
GGCTCTCCTC
AAGCGTATTC
AACAAGGGGC
TGAAGGATGC
CCAGAAGGTA
CCCCATTGTA
TGGGATCTGA
TCTGGGGCCT
CGGTGCACAT
GCTTTACATG
TGTTTAGTCG
AGGTTAAAAA
AACGTCTAGG
CCCCCCGAAC
CACGGGGACG
TGGTTTTCCT
TTGAAAAACA
CGATGATAAG
CTTGCCACAA
CCCACAAGGA
GACGACCTTC
CATGACCGAG
TACAAGCCCA
CGGTGCGCCT
CGCCACCCGC
GACGACGTCC
CCCGGGCCGT
ACGCACCCTC
GCCGCCGCGT
TCGCCGACTA
CCCCGCCACG
CGCCACACCG
TCGACCCGGA
CCGCCACATC
GAGCGGGTCA
CCGAGCTGCA
AGAACTCTTC
CTCACGCGCG
TCGGGCTCGA
CATCGGCAAG
GTGTGGGTCG
CGGACGACGG
CGCCGCGGTG
GCGGTCTGGA
CCACGCCGGA
GAGCGTCGAA
GCGGGGGCGG
TGTTCGCCGA
GATCGGCCCG
CGCATGGCCG
AGTTGAGCGG
TTCCCGGCTG
GCCGCGCAGC
AACAGATGGA
AGGCCTCCTG
GCGCCGCACC
GGCCCAAGGA
GCCCGCGTGG
TTCCTGGCCA
CCGTCGGCGT
CTCGCCCGAC
CACCAGGGCA
AGGGTCTGGG
CAGCGCCGTC
GTGCTCCCCG
GAGTGGAGGC
GGCCGAGCGC
GCCGGGGTGC
CCGCCTTCCT
GGAGACCTCC
GCGCCCCGCA
ACCTCCCCTT
CTACGAGCGG
CTCGGCTTCA
CCGTCACCGC
CGACGTCGAG
GTGCCCGAAG
GACCGCGCAC
CTGGTGCATG
ACCCGCAAGC
CCGGTGCCTG
ATAATAGGCG
GCCGCTCGAG
ACCTAGAAAA
ACATGGAGCA
ATCACAAGTA
GCAATACAGC
AGCTACCAAT
GCTGATTGTG
CCTGGCTAGA
AGCACAAGAG
GAGGAGGAGG
TGGGTTTTCC
AGTCACACCT
CAGGTACCTT
TAAGACCAAT
GACTTACAAG
GCAGCTGTAG
ATCTTAGCCA
CTTTTTAAAA
GAAAAGGGGG
GACTGGAAGG
GCTAATTCAC
TCCCAACGAA
GACAAGATCT
GCTTTTTGCT
TGTACTGGGT
CTCTCTGGTT
AGACCAGATC
TGAGCCTGGG
AGCTCTCTGG
CTAACTAGGG
AACCCACTGC
TTAAGCCTCA
ATAAAGCTTG
CCTTGAGTGC
TTCAAGTAGT
GTGTGCCCGT
CTGTTGTGTG
ACTCTGGTAA
CTAGAGATCC
CTCAGACCCT
TTTAGTCAGT
GTGGAAAATC
TCTAGCAGTA
GTAGTTCATG
TCATCTTATT
ATTCAGTATT
TATAACTTGC
AAAGAAATGA
ATATCAGAGA
GTGAGAGGCC
TTGACATTAT
AATAGATTTA
GCAGGAATTG
AACTAGGAGT
GGAGCACACA
GGCAAAGTTC
TAGAGCTCGC
TGATCAGCCT
CGACTGTGCC
TTCTAGTTGC
CAGCCATCTG
TTGTTTGCCC
CTCCCCCGTG
CCTTCCTTGA
CCCTGGAAGG
TGCCACTCCC
ACTGTCCTTT
CCTAATAAAA
TGAGGAAATT
GCATCGCATT
GTCTGAGTAG
GTGTCATTCT
ATTCTGGGGG
GTGGGGGTGG
GGCAGGACAG
CAAGGGGGGA
GGATTGGGAA
GACAATAGCA
GGCATGCTGG
GGATGCGGTG
GGCTCTATGG
CTTCTGAGGC
GGAAAGAACC
AGCTGGGGGC
GCGCCCCTCG
AGGCCGCCAT
GGTCATAGCT
GTTTGACGTC
AGGTGGCACT
TTTCGGGGAA
ATGTGCGCGG
AACCCCTATT
TGTTTATTTT
TCTAAATACA
TTCAAATATG
TATCCGCTCA
CGAGACAATA
ACCCTGATAA
ATGCTTCAAT
AATATTGAAA
AAGGAAGAGT
ATGAGTATTC
AACATTTCCG
TGTCGCCCTT
ATTCCCTTTT
TTGCGGCATT
TTGCCTTCCT
GTTTTTGCTC
ACCCAGAAAC
GCTGGTGAAA
GTAAAAGATG
CTGAAGATCA
GTTGGGTGCA
CGAGTGGGTT
ACATCGAACT
GGATCTCAAC
AGCGGTAAGA
TCCTTGAGAG
TTTTCGCCCC
GAAGAACGTT
TTCCAATGAT
GAGCACTTTT
AAAGTTCTGC
TATGTGGCGC
GGTATTATCC
CGTATTGACG
CCGGGCAAGA
GCAACTCGGT
CGCCGCATAC
ACTATTCTCA
GAATGACTTG
GTTGAGTACT
CACCAGTCAC
AGAAAAGCAT
CTTACGGATG
GCATGACAGT
AAGAGAATTA
TGCAGTGCTG
CCATAACCAT
GAGTGATAAC
ACTGCGGCCA
ACTTACTTCT
GACAACGATC
GGAGGACCGA
AGGAGCTAAC
CGCTTTTTTG
CACAACATGG
GGGATCATGT
AACTCGCCTT
GATCGTTGGG
AACCGGAGCT
GAATGAAGCC
ATACCAAACG
ACGAGCGTGA
CACCACGATG
CCTGTAGCAA
TGGCAACAAC
GTTGCGCAAA
CTATTAACTG
GCGAACTACT
TACTCTAGCT
TCCCGGCAAC
AATTAATAGA
CTGGATGGAG
GCGGATAAAG
TTGCAGGACC
ACTTCTGCGC
TCGGCCCTTC
CGGCTGGCTG
GTTTATTGCT
GATAAATCTG
GAGCCGGTGA
GCGTGGGTCT
CGCGGTATCA
TTGCAGCACT
GGGGCCAGAT
GGTAAGCCCT
CCCGTATCGT
AGTTATCTAC
ACGACGGGGA
GTCAGGCAAC
TATGGATGAA
CGAAATAGAC
AGATCGCTGA
GATAGGTGCC
TCACTGATTA
AGCATTGGTA
ACTGTCAGAC
CAAGTTTACT
CATATATACT
TTAGATTGAT
TTAAAACTTC
ATTTTTAATT
TAAAAGGATC
TAGGTGAAGA
TCCTTTTTGA
TAATCTCATG
ACCAAAATCC
CTTAACGTGA
GTTTTCGTTC
CACTGAGCGT
CAGACCCCGT
AGAAAAGATC
AAAGGATCTT
CTTGAGATCC
TTTTTTTCTG
CGCGTAATCT
GCTGCTTGCA
AACAAAAAAA
CCACCGCTAC
CAGCGGTGGT
TTGTTTGCCG
GATCAAGAGC
TACCAACTCT
TTTTCCGAAG
GTAACTGGCT
TCAGCAGAGC
GCAGATACCA
AATACTGTTC
TTCTAGTGTA
GCCGTAGTTA
GGCCACCACT
TCAAGAACTC
TGTAGCACCG
CCTACATACC
TCGCTCTGCT
AATCCTGTTA
CCAGTGGCTG
CTGCCAGTGG
CGATAAGTCG
TGTCTTACCG
GGTTGGACTC
AAGACGATAG
TTACCGGATA
AGCGCAGCGG
TCGGGCTGAA
CGGGGGGTTC
GTGCACACAG
CCCAGCTTGG
AGCGAACGAC
CTACACCGAA
CTGAGATACC
TACAGCGTGA
GCTATGAGAA
AGCGCCACGC
TTCCCGAAGG
GAGAAAGGCG
GACAGGTATC
CGGTAAGCGG
CAGGGTCGGA
ACAGGAGAGC
GCACGAGGGA
GCTTCCAGGG
GGAAACGCCT
GGTATCTTTA
TAGTCCTGTC
GGGTTTCGCC
ACCTCTGACT
TGAGCGTCGA
TTTTTGTGAT
GCTCGTCAGG
GGGGCGGAGC
CTATGGAAAA
ACGCCAGCAA
CGCGGCCTTT
TTACGGTTCT
GGCCTTTTGC
A