名称: | pSIN-EF2-MCS-P2A-EGFP-Neo |
---|---|
别称: | - |
基因长: | |
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: | 空载体 |
原核抗性: | Amp |
筛选标记: | Neo/G418 |
荧光标记: | 绿色 |
启动子: | EF1a |
复制子: | pUC |
拷贝数: | |
感受态: | Stbl3 |
温度: | 37度 |
背景载体: | pSIN-Neo |
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.
pSIN-EF2-MCS-P2A-EGFP-Neo 是一种基于慢病毒的表达载体,质粒携带的EF1a启动子可启动下游基因的表达,质粒携带 EGFP绿色荧光蛋白,可用于检测目的基因表达情况和位置。质粒携带Neo/G418抗性基因,用于筛选表达目的基因的宿主细胞。
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
ACTTTGGTCG
ATAAGCTTTG
CAAAGATGGA
TAAAGTTTTA
AACAGAGAGG
AATCTTTGCA
GCTAATGGAC
CTTCTAGGTC
TTGAAAGGAG
TGGGAATTGG
CTCCGGTGCC
CGTCAGTGGG
CAGAGCGCAC
ATCGCCCACA
GTCCCCGAGA
AGTTGGGGGG
AGGGGTCGGC
AATTGAACCG
GTGCCTAGAG
AAGGTGGCGC
GGGGTAAACT
GGGAAAGTGA
TGTCGTGTAC
TGGCTCCGCC
TTTTTCCCGA
GGGTGGGGGA
GAACCGTATA
TAAGTGCAGT
AGTCGCCGTG
AACGTTCTTT
TTCGCAACGG
GTTTGCCGCC
AGAACACAGG
TAAGTGCCGT
GTGTGGTTCC
CGCGGGCCTG
GCCTCTTTAC
GGGTTATGGC
CCTTGCGTGC
CTTGAATTAC
TTCCACCTGG
CTGCAGTACG
TGATTCTTGA
TCCCGAGCTT
CGGGTTGGAA
GTGGGTGGGA
GAGTTCGAGG
CCTTGCGCTT
AAGGAGCCCC
TTCGCCTCGT
GCTTGAGTTG
AGGCCTGGCC
TGGGCGCTGG
GGCCGCCGCG
TGCGAATCTG
GTGGCACCTT
CGCGCCTGTC
TCGCTGCTTT
CGATAAGTCT
CTAGCCATTT
AAAATTTTTG
ATGACCTGCT
GCGACGCTTT
TTTTCTGGCA
AGATAGTCTT
GTAAATGCGG
GCCAAGATCT
GCACACTGGT
ATTTCGGTTT
TTGGGGCCGC
GGGCGGCGAC
GGGGCCCGTG
CGTCCCAGCG
CACATGTTCG
GCGAGGCGGG
GCCTGCGAGC
GCGGCCACCG
AGAATCGGAC
GGGGGTAGTC
TCAAGCTGGC
CGGCCTGCTC
TGGTGCCTGG
CCTCGCGCCG
CCGTGTATCG
CCCCGCCCTG
GGCGGCAAGG
CTGGCCCGGT
CGGCACCAGT
TGCGTGAGCG
GAAAGATGGC
CGCTTCCCGG
CCCTGCTGCA
GGGAGCTCAA
AATGGAGGAC
GCGGCGCTCG
GGAGAGCGGG
CGGGTGAGTC
ACCCACACAA
AGGAAAAGGG
CCTTTCCGTC
CTCAGCCGTC
GCTTCATGTG
ACTCCACGGA
GTACCGGGCG
CCGTCCAGGC
ACCTCGATTA
GTTCTCGAGC
TTTTGGAGTA
CGTCGTCTTT
AGGTTGGGGG
GAGGGGTTTT
ATGCGATGGA
GTTTCCCCAC
ACTGAGTGGG
TGGAGACTGA
AGTTAGGCCA
GCTTGGCACT
TGATGTAATT
CTCCTTGGAA
TTTGCCCTTT
TTGAGTTTGG
ATCTTGGTTC
ATTCTCAAGC
CTCAGACAGT
GGTTCAAAGT
TTTTTTCTTC
CATTTCAGGT
GTCGTGAGGA
ATTGGGATCC
TTCGAACATA
TGGCTAGCGA
ATTCGCCACT
AACTTCTCCC
TGTTGAAACA
AGCAGGGGAT
GTCGAAGAGA
ATCCCGGGCC
ACCGGTCGCC
ACCATGGTGA
GCAAGGGCGA
GGAGCTGTTC
ACCGGGGTGG
TGCCCATCCT
GGTCGAGCTG
GACGGCGACG
TAAACGGCCA
CAAGTTCAGC
GTGTCCGGCG
AGGGCGAGGG
CGATGCCACC
TACGGCAAGC
TGACCCTGAA
GTTCATCTGC
ACCACCGGCA
AGCTGCCCGT
GCCCTGGCCC
ACCCTCGTGA
CCACCCTGAC
CTACGGCGTG
CAGTGCTTCA
GCCGCTACCC
CGACCACATG
AAGCAGCACG
ACTTCTTCAA
GTCCGCCATG
CCCGAAGGCT
ACGTCCAGGA
GCGCACCATC
TTCTTCAAGG
ACGACGGCAA
CTACAAGACC
CGCGCCGAGG
TGAAGTTCGA
GGGCGACACC
CTGGTGAACC
GCATCGAGCT
GAAGGGCATC
GACTTCAAGG
AGGACGGCAA
CATCCTGGGG
CACAAGCTGG
AGTACAACTA
CAACAGCCAC
AACGTCTATA
TCATGGCCGA
CAAGCAGAAG
AACGGCATCA
AGGTGAACTT
CAAGATCCGC
CACAACATCG
AGGACGGCAG
CGTGCAGCTC
GCCGACCACT
ACCAGCAGAA
CACCCCCATC
GGCGACGGCC
CCGTGCTGCT
GCCCGACAAC
CACTACCTGA
GCACCCAGTC
CGCCCTGAGC
AAAGACCCCA
ACGAGAAGCG
CGATCACATG
GTCCTGCTGG
AGTTCGTGAC
CGCCGCCGGG
ATCACTCTCG
GCATGGACGA
GCTGTACAAG
TAACTAGTGA
TCCGCATGCA
TCTAGGGCGG
CCAATTCCGC
CCCTCTCCCT
CCCCCCCCCC
TAACGTTACT
GGCCGAAGCC
GCTTGGAATA
AGGCCGGTGT
GCGTTTGTCT
ATATGTGATT
TTCCACCATA
TTGCCGTCTT
TTGGCAATGT
GAGGGCCCGG
AAACCTGGCC
CTGTCTTCTT
GACGAGCATT
CCTAGGGGTC
TTTCCCCTCT
CGCCAAAGGA
ATGCAAGGTC
TGTTGAATGT
CGTGAAGGAA
GCAGTTCCTC
TGGAAGCTTC
TTGAAGACAA
ACAACGTCTG
TAGCGACCCT
TTGCAGGCAG
CGGAACCCCC
CACCTGGCGA
CAGGTGCCTC
TGCGGCCAAA
AGCCACGTGT
ATAAGATACA
CCTGCAAAGG
CGGCACAACC
CCAGTGCCAC
GTTGTGAGTT
GGATAGTTGT
GGAAAGAGTC
AAATGGCTCT
CCTCAAGCGT
ATTCAACAAG
GGGCTGAAGG
ATGCCCAGAA
GGTACCCCAT
TGTATGGGAT
CTGATCTGGG
GCCTCGGTGC
ACATGCTTTA
CATGTGTTTA
GTCGAGGTTA
AAAAAACGTC
TAGGCCCCCC
GAACCACGGG
GACGTGGTTT
TCCTTTGAAA
AACACGATGA
TAAGCTTGCC
ACAACCCACA
AGGAGACGAC
CTTCCATGAT
TGAACAAGAT
GGATTGCACG
CAGGTTCTCC
GGCCGCTTGG
GTGGAGAGGC
TATTCGGCTA
TGACTGGGCA
CAACAGACAA
TCGGCTGCTC
TGATGCCGCC
GTGTTCCGGC
TGTCAGCGCA
GGGGCGCCCG
GTTCTTTTTG
TCAAGACCGA
CCTGTCCGGT
GCCCTGAATG
AACTGCAGGA
CGAGGCAGCG
CGGCTATCGT
GGCTGGCCAC
GACGGGCGTT
CCTTGCGCAG
CTGTGCTCGA
CGTTGTCACT
GAAGCGGGAA
GGGACTGGCT
GCTATTGGGC
GAAGTGCCGG
GGCAGGATCT
CCTGTCATCT
CACCTTGCTC
CTGCCGAGAA
AGTATCCATC
ATGGCTGATG
CAATGCGGCG
GCTGCATACG
CTTGATCCGG
CTACCTGCCC
ATTCGACCAC
CAAGCGAAAC
ATCGCATCGA
GCGAGCACGT
ACTCGGATGG
AAGCCGGTCT
TGTCGATCAG
GATGATCTGG
ACGAAGAGCA
TCAGGGGCTC
GCGCCAGCCG
AACTGTTCGC
CAGGCTCAAG
GCGCGCATGC
CCGACGGCGA
GGATCTCGTC
GTGACCCATG
GCGATGCCTG
CTTGCCGAAT
ATCATGGTGG
AAAATGGCCG
CTTTTCTGGA
TTCATCGACT
GTGGCCGGCT
GGGTGTGGCG
GACCGCTATC
AGGACATAGC
GTTGGCTACC
CGTGATATTG
CTGAAGAGCT
TGGCGGCGAA
TGGGCTGACC
GCTTCCTCGT
GCTTTACGGT
ATCGCCGCTC
CCGATTCGCA
GCGCATCGCC
TTCTATCGCC
TTCTTGACGA
GTTCTTCTGA
TAATAGGCGG
CCGCTCGAGA
CCTAGAAAAA
CATGGAGCAA
TCACAAGTAG
CAATACAGCA
GCTACCAATG
CTGATTGTGC
CTGGCTAGAA
GCACAAGAGG
AGGAGGAGGT
GGGTTTTCCA
GTCACACCTC
AGGTACCTTT
AAGACCAATG
ACTTACAAGG
CAGCTGTAGA
TCTTAGCCAC
TTTTTAAAAG
AAAAGGGGGG
ACTGGAAGGG
CTAATTCACT
CCCAACGAAG
ACAAGATCTG
CTTTTTGCTT
GTACTGGGTC
TCTCTGGTTA
GACCAGATCT
GAGCCTGGGA
GCTCTCTGGC
TAACTAGGGA
ACCCACTGCT
TAAGCCTCAA
TAAAGCTTGC
CTTGAGTGCT
TCAAGTAGTG
TGTGCCCGTC
TGTTGTGTGA
CTCTGGTAAC
TAGAGATCCC
TCAGACCCTT
TTAGTCAGTG
TGGAAAATCT
CTAGCAGTAG
TAGTTCATGT
CATCTTATTA
TTCAGTATTT
ATAACTTGCA
AAGAAATGAA
TATCAGAGAG
TGAGAGGCCT
TGACATTATA
ATAGATTTAG
CAGGAATTGA
ACTAGGAGTG
GAGCACACAG
GCAAAGTTCT
AGAGCTCGCT
GATCAGCCTC
GACTGTGCCT
TCTAGTTGCC
AGCCATCTGT
TGTTTGCCCC
TCCCCCGTGC
CTTCCTTGAC
CCTGGAAGGT
GCCACTCCCA
CTGTCCTTTC
CTAATAAAAT
GAGGAAATTG
CATCGCATTG
TCTGAGTAGG
TGTCATTCTA
TTCTGGGGGG
TGGGGTGGGG
CAGGACAGCA
AGGGGGAGGA
TTGGGAAGAC
AATAGCAGGC
ATGCTGGGGA
TGCGGTGGGC
TCTATGGCTT
CTGAGGCGGA
AAGAACCAGC
TGGGGGCGCG
CCCCTCGAGG
CCGCCATGGT
CATAGCTGTT
TGACGTCAGG
TGGCACTTTT
CGGGGAAATG
TGCGCGGAAC
CCCTATTTGT
TTATTTTTCT
AAATACATTC
AAATATGTAT
CCGCTCATGA
GACAATAACC
CTGATAAATG
CTTCAATAAT
ATTGAAAAAG
GAAGAGTATG
AGTATTCAAC
ATTTCCGTGT
CGCCCTTATT
CCCTTTTTTG
CGGCATTTTG
CCTTCCTGTT
TTTGCTCACC
CAGAAACGCT
GGTGAAAGTA
AAAGATGCTG
AAGATCAGTT
GGGTGCACGA
GTGGGTTACA
TCGAACTGGA
TCTCAACAGC
GGTAAGATCC
TTGAGAGTTT
TCGCCCCGAA
GAACGTTTTC
CAATGATGAG
CACTTTTAAA
GTTCTGCTAT
GTGGCGCGGT
ATTATCCCGT
ATTGACGCCG
GGCAAGAGCA
ACTCGGTCGC
CGCATACACT
ATTCTCAGAA
TGACTTGGTT
GAGTACTCAC
CAGTCACAGA
AAAGCATCTT
ACGGATGGCA
TGACAGTAAG
AGAATTATGC
AGTGCTGCCA
TAACCATGAG
TGATAACACT
GCGGCCAACT
TACTTCTGAC
AACGATCGGA
GGACCGAAGG
AGCTAACCGC
TTTTTTGCAC
AACATGGGGG
ATCATGTAAC
TCGCCTTGAT
CGTTGGGAAC
CGGAGCTGAA
TGAAGCCATA
CCAAACGACG
AGCGTGACAC
CACGATGCCT
GTAGCAATGG
CAACAACGTT
GCGCAAACTA
TTAACTGGCG
AACTACTTAC
TCTAGCTTCC
CGGCAACAAT
TAATAGACTG
GATGGAGGCG
GATAAAGTTG
CAGGACCACT
TCTGCGCTCG
GCCCTTCCGG
CTGGCTGGTT
TATTGCTGAT
AAATCTGGAG
CCGGTGAGCG
TGGGTCTCGC
GGTATCATTG
CAGCACTGGG
GCCAGATGGT
AAGCCCTCCC
GTATCGTAGT
TATCTACACG
ACGGGGAGTC
AGGCAACTAT
GGATGAACGA
AATAGACAGA
TCGCTGAGAT
AGGTGCCTCA
CTGATTAAGC
ATTGGTAACT
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
ATTGAGAAAG
CGCCACGCTT
CCCGAAGGGA
GAAAGGCGGA
CAGGTATCCG
GTAAGCGGCA
GGGTCGGAAC
AGGAGAGCGC
ACGAGGGAGC
TTCCAGGGGG
AAACGCCTGG
TATCTTTATA
GTCCTGTCGG
GTTTCGCCAC
CTCTGACTTG
AGCGTCGATT
TTTGTGATGC
TCGTCAGGGG
GGCGGAGCCT
ATGGAAAAAC
GCCAGCAACG
CGGCCTTTTT
ACGGTTCCTG
GCCTTTTGC