pYES2-NTA
pYES2-NTA
pYES2-NTA
编号 | 载体名称 |
北京华越洋生物VECT2230 | pYES2-NTA |
pYES2-NTA载体基本信息
| 载体名称: | pYES2-NTA, pYES2/NT A |
|---|---|
| 质粒类型: | 酿酒酵母蛋白表达载体 |
| 表达水平: | 高拷贝 |
| 诱导方法: | 半乳糖 |
| 启动子: | GAL1 |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 6038 bp |
| 5' 测序引物及序列: | T7: TAATACGACTCACTATAGGG |
| 3' 测序引物及序列: | CYC1 Terminator: GTGACATAACTAATTACATGATG |
| 载体标签: | N-His, N-Xpress, C-His, C-V5 |
| 载体抗性: | 氨苄 |
| 筛选标记: | URA3 |
| 备注: | 利用半乳糖诱导蛋白在酿酒酵母中表达。 |
| 稳定性: | 稳定 Stable |
| 组成型/诱导型: | 诱导型 |
| 病毒/非病毒: | 非病毒 |
pYES2-NTA载体质粒图谱和多克隆位点信息
pYES2-NTA载体简介
pYES2 NT A, B, C的是一个6.0 kb的载体，设计用来在酿酒酵母（Saccharomyces cerevisiae）中诱导表达重组蛋白。
该载体包含以下元素：
1. 酵母GAL1启动子，能够在酿酒酵母中被半乳糖高水平的诱导蛋白表达目的蛋白，同时能够被葡萄糖抑制表达
2.多克隆位点可以使用的很多限制酶切位点，便于基因插入。
3.CYC1终止子能够有效终止mRNA的转录。
4.能够利用URA3基因筛选带有ura3基因型的酵母宿主菌株转化子。
5.氨苄抗性基因能够方便在大肠杆菌中的进行载体筛选。
6. 载体本身带有N端His和Xpress标签，同时载体C端也带有V5抗原表位和His纯化标签。
8. 在酵母中，利用载体上的2μ复制子维持质粒的高拷贝。
pYES2-NTA载体序列
LOCUS pYES2/NTA 6038 bp DNA circular SYN DEFINITION pYES2/NTA ACCESSION KEYWORDS SOURCE ORGANISM other sequences; artificial sequences; vectors. COMMENT This file is created by Vector NTI FEATURES Location/Qualifiers source 1..6038 /organism="pYES2/NT A" /mol_type="other DNA" promoter 1..451 /label="GAL1_promoter" misc_feature 414..437 /label="GAL1_primer" promoter 475..493 /label="T7_promoter" misc_feature 542..560 /label="Xpress_fwd_primer" misc_feature 543..575 /label="T7_leader" misc_feature 579..602 /label="Xpress_EK" terminator 791..1036 /label="CYC1_terminator" misc_feature complement(800..818) /label="CYC1_primer" misc_feature complement(800..818) /label="pYESTrp_rev_primer" rep_origin complement(1264..1883) /label="pBR322_origin" CDS complement(2038..2898) /label="ORF frame 3" CDS complement(2994..3797) /label="ORF frame 1" gene complement(2997..3797) /label="URA3" /gene="URA3" CDS 3286..3762 /label="ORF frame 1" promoter complement(3799..4024) /label="URA3_promoter" rep_origin 4027..5498 /label="2micron_origin" ORIGIN 1 ACGGATTAGA AGCCGCCGAG CGGGTGACAG CCCTCCGAAG GAAGACTCTC CTCCGTGCGT 61 CCTCGTCTTC ACCGGTCGCG TTCCTGAAAC GCAGATGTGC CTCGCGCCGC ACTGCTCCGA 121 ACAATAAAGA TTCTACAATA CTAGCTTTTA TGGTTATGAA GAGGAAAAAT TGGCAGTAAC 181 CTGGCCCCAC AAACCTTCAA ATGAACGAAT CAAATTAACA ACCATAGGAT GATAATGCGA 241 TTAGTTTTTT AGCCTTATTT CTGGGGTAAT TAATCAGCGA AGCGATGATT TTTGATCTAT 301 TAACAGATAT ATAAATGCAA AAACTGCATA ACCACTTTAA CTAATACTTT CAACATTTTC 361 GGTTTGTATT ACTTCTTATT CAAATGTAAT AAAAGTATCA ACAAAAAATT GTTAATATAC 421 CTCTATACTT TAACGTCAAG GAGAAAAAAC CCCGGATCGG ACTACTAGCA GCTGTAATAC 481 GACTCACTAT AGGGAATATT AAGCTTACCA TGGGGGGTTC TCATCATCAT CATCATCATG 541 GTATGGCTAG CATGACTGGT GGACAGCAAA TGGGTCGGGA TCTGTACGAC GATGACGATA 601 AGGTACCTAG GATCCAGTGT GGTGGAATTC TGCAGATATC CAGCACAGTG GCGGCCGCTC 661 GAGTCTAGAG GGCCCTTCGA AGGTAAGCCT ATCCCTAACC CTCTCCTCGG TCTCGATTCT 721 ACGCGTACCG GTCATCATCA CCATCACCAT TGAGTTTAAA CCCGCTGATC CTAGAGGGCC 781 GCATCATGTA ATTAGTTATG TCACGCTTAC ATTCACGCCC TCCCCCCACA TCCGCTCTAA 841 CCGAAAAGGA AGGAGTTAGA CAACCTGAAG TCTAGGTCCC TATTTATTTT TTTATAGTTA 901 TGTTAGTATT AAGAACGTTA TTTATATTTC AAATTTTTCT TTTTTTTCTG TACAGACGCG 961 TGTACGCATG TAACATTATA CTGAAAACCT TGCTTGAGAA GGTTTTGGGA CGCTCGAAGG 1021 CTTTAATTTG CAAGCTGCGG CCCTGCATTA ATGAATCGGC CAACGCGCGG GGAGAGGCGG 1081 TTTGCGTATT GGGCGCTCTT CCGCTTCCTC GCTCACTGAC TCGCTGCGCT CGGTCGTTCG 1141 GCTGCGGCGA GCGGTATCAG CTCACTCAAA GGCGGTAATA CGGTTATCCA CAGAATCAGG 1201 GGATAACGCA GGAAAGAACA TGTGAGCAAA AGGCCAGCAA AAGCCCAGGA ACCGTAAAAA 1261 GGCCGCGTTG CTGGCGTTTT TCCATAGGCT CCGCCCCCCT GACGAGCATC ACAAAAATCG 1321 ACGCTCAAGT CAGAGGTGGC GAAACCCGAC AGGACTATAA AGATACCAGG CGTTTCCCCC 1381 TGGAAGCTCC CTCGTGCGCT CTCCTGTTCC GACCCTGCCG CTTACCGGAT ACCTGTCCGC 1441 CTTTCTCCCT TCGGGAAGCG TGGCGCTTTC TCATAGCTCA CGCTGTAGGT ATCTCAGTTC 1501 GGTGTAGGTC GTTCGCTCCA AGCTGGGCTG TGTGCACGAA CCCCCCGTTC AGCCCGACCG 1561 CTGCGCCTTA TCCGGTAACT ATCGTCTTGA GTCCAACCCG GTAAGACACG ACTTATCGCC 1621 ACTGGCAGCA GCCACTGGTA ACAGGATTAG CAGAGCGAGG TATGTAGGCG GTGCTACAGA 1681 GTTCTTGAAG TGGTGGCCTA ACTACGGCTA CACTAGAAGG ACAGTATTTG GTATCTGCGC 1741 TCTGCTGAAG CCAGTTACCT TCGGAAAAAG AGTTGGTAGC TCTTGATCCG GCAAACAAAC 1801 CACCGCTGGT AGCGGTGGTT TTTTTGTTTG CAAGCAGCAG ATTACGCGCA GAAAAAAAGG 1861 ATCTCAAGAA GATCCTTTGA TCTTTTCTAC GGGGTCTGAC GCTCAGTGGA ACGAAAACTC 1921 ACGTTAAGGG ATTTTGGTCA TGAGATTATC AAAAAGGATC TTCACCTAGA TCCTTTTAAA 1981 TTAAAAATGA AGTTTTAAAT CAATCTAAAG TATATATGAG TAAACTTGGT CTGACAGTTA 2041 CCAATGCTTA ATCAGTGAGG CACCTATCTC AGCGATCTGT CTATTTCGTT CATCCATAGT 2101 TGCCTGACTC CCCGTCGTGT AGATAACTAC GATACGGGAG CGCTTACCAT CTGGCCCCAG 2161 TGCTGCAATG ATACCGCGAG ACCCACGCTC ACCGGCTCCA GATTTATCAG CAATAAACCA 2221 GCCAGCCGGA AGGGCCGAGC GCAGAAGTGG TCCTGCAACT TTATCCGCCT CCATCCAGTC 2281 TATTAATTGT TGCCGGGAAG CTAGAGTAAG TAGTTCGCCA GTTAATAGTT TGCGCAACGT 2341 TGTTGGCATT GCTACAGGCA TCGTGGTGTC ACTCTCGTCG TTTGGTATGG CTTCATTCAG 2401 CTCCGGTTCC CAACGATCAA GGCGAGTTAC ATGATCCCCC ATGTTGTGCA AAAAAGCGGT 2461 TAGCTCCTTC GGTCCTCCGA TCGTTGTCAG AAGTAAGTTG GCCGCAGTGT TATCACTCAT 2521 GGTTATGGCA GCACTGCATA ATTCTCTTAC TGTCATGCCA TCCGTAAGAT GCTTTTCTGT 2581 GACTGGTGAG TACTCAACCA AGTCATTCTG AGAATAGTGT ATGCGGCGAC CGAGTTGCTC 2641 TTGCCCGGCG TCAATACGGG ATAATAGTGT ATCACATAGC AGAACTTTAA AAGTGCTCAT 2701 CATTGGAAAA CGTTCTTCGG GGCGAAAACT CTCAAGGATC TTACCGCTGT TGAGATCCAG 2761 TTCGATGTAA CCCACTCGTG CACCCAACTG ATCTTCAGCA TCTTTTACTT TCACCAGCGT 2821 TTCTGGGTGA GCAAAAACAG GAAGGCAAAA TGCCGCAAAA AAGGGAATAA GGGCGACACG 2881 GAAATGTTGA ATACTCATAC TCTTCCTTTT TCAATGGGTA ATAACTGATA TAATTAAATT 2941 GAAGCTCTAA TTTGTGAGTT TAGTATACAT GCATTTACTT ATAATACAGT TTTTTAGTTT 3001 TGCTGGCCGC ATCTTCTCAA ATATGCTTCC CAGCCTGCTT TTCTGTAACG TTCACCCTCT 3061 ACCTTAGCAT CCCTTCCCTT TGCAAATAGT CCTCTTCCAA CAATAATAAT GTCAGATCCT 3121 GTAGAGACCA CATCATCCAC GGTTCTATAC TGTTGACCCA ATGCGTCTCC CTTGTCATCT 3181 AAACCCACAC CGGGTGTCAT AATCAACCAA TCGTAACCTT CATCTCTTCC ACCCATGTCT 3241 CTTTGAGCAA TAAAGCCGAT AACAAAATCT TTGTCGCTCT TCGCAATGTC AACAGTACCC 3301 TTAGTATATT CTCCAGTAGA TAGGGAGCCC TTGCATGACA ATTCTGCTAA CATCAAAAGG 3361 CCTCTAGGTT CCTTTGTTAC TTCTTCTGCC GCCTGCTTCA AACCGCTAAC AATACCTGGG 3421 CCCACCACAC CGTGTGCATT CGTAATGTCT GCCCATTCTG CTATTCTGTA TACACCCGCA 3481 GAGTACTGCA ATTTGACTGT ATTACCAATG TCAGCAAATT TTCTGTCTTC GAAGAGTAAA 3541 AAATTGTACT TGGCGGATAA TGCCTTTAGC GGCTTAACTG TGCCCTCCAT GGAAAAATCA 3601 GTCAAGATAT CCACATGTGT TTTTAGTAAA CAAATTTTGG GACCTAATGC TTCAACTAAC 3661 TCCAGTAATT CCTTGGTGGT ACGAACATCC AATGAAGCAC ACAAGTTTGT TTGCTTTTCG 3721 TGCATGATAT TAAATAGCTT GGCAGCAACA GGACTAGGAT GAGTAGCAGC ACGTTCCTTA 3781 TATGTAGCTT TCGACATGAT TTATCTTCGT TTCCTGCAGG TTTTTGTTCT GTGCAGTTGG 3841 GTTAAGAATA CTGGGCAATT TCATGTTTCT TCAACACTAC ATATGCGTAT ATATACCAAT 3901 CTAAGTCTGT GCTCCTTCCT TCGTTCTTCC TTCTGTTCGG AGATTACCGA ATCAAAAAAA 3961 TTTCAAAGAA ACCGAAATCA AAAAAAAGAA TAAAAAAAAA ATGATGAATT GAATTGAAAA 4021 GCTAGCTTAT CGATGATAAG CTGTCAAAGA TGAGAATTAA TTCCACGGAC TATAGACTAT 4081 ACTAGATACT CCGTCTACTG TACGATACAC TTCCGCTCAG GTCCTTGTCC TTTAACGAGG 4141 CCTTACCACT CTTTTGTTAC TCTATTGATC CAGCTCAGCA AAGGCAGTGT GATCTAAGAT 4201 TCTATCTTCG CGATGTAGTA AAACTAGCTA GACCGAGAAA GAGACTAGAA ATGCAAAAGG 4261 CACTTCTACA ATGGCTGCCA TCATTATTAT CCGATGTGAC GCTGCAGCTT CTCAATGATA 4321 TTCGAATACG CTTTGAGGAG ATACAGCCTA ATATCCGACA AACTGTTTTA CAGATTTACG 4381 ATCGTACTTG TTACCCATCA TTGAATTTTG AACATCCGAA CCTGGGAGTT TTCCCTGAAA 4441 CAGATAGTAT ATTTGAACCT GTATAATAAT ATATAGTCTA GCGCTTTACG GAAGACAATG 4501 TATGTATTTC GGTTCCTGGA GAAACTATTG CATCTATTGC ATAGGTAATC TTGCACGTCG 4561 CATCCCCGGT TCATTTTCTG CGTTTCCATC TTGCACTTCA ATAGCATATC TTTGTTAACG 4621 AAGCATCTGT GCTTCATTTT GTAGAACAAA AATGCAACGC GAGAGCGCTA ATTTTTCAAA 4681 CAAAGAATCT GAGCTGCATT TTTACAGAAC AGAAATGCAA CGCGAAAGCG CTATTTTACC 4741 AACGAAGAAT CTGTGCTTCA TTTTTGTAAA ACAAAAATGC AACGCGACGA GAGCGCTAAT 4801 TTTTCAAACA AAGAATCTGA GCTGCATTTT TACAGAACAG AAATGCAACG CGAGAGCGCT 4861 ATTTTACCAA CAAAGAATCT ATACTTCTTT TTTGTTCTAC AAAAATGCAT CCCGAGAGCG 4921 CTATTTTTCT AACAAAGCAT CTTAGATTAC TTTTTTTCTC CTTTGTGCGC TCTATAATGC 4981 AGTCTCTTGA TAACTTTTTG CACTGTAGGT CCGTTAAGGT TAGAAGAAGG CTACTTTGGT 5041 GTCTATTTTC TCTTCCATAA AAAAAGCCTG ACTCCACTTC CCGCGTTTAC TGATTACTAG 5101 CGAAGCTGCG GGTGCATTTT TTCAAGATAA AGGCATCCCC GATTATATTC TATACCGATG 5161 TGGATTGCGC ATACTTTGTG AACAGAAAGT GATAGCGTTG ATGATTCTTC ATTGGTCAGA 5221 AAATTATGAA CGGTTTCTTC TATTTTGTCT CTATATACTA CGTATAGGAA ATGTTTACAT 5281 TTTCGTATTG TTTTCGATTC ACTCTATGAA TAGTTCTTAC TACAATTTTT TTGTCTAAAG 5341 AGTAATACTA GAGATAAACA TAAAAAATGT AGAGGTCGAG TTTAGATGCA AGTTCAAGGA 5401 GCGAAAGGTG GATGGGTAGG TTATATAGGG ATATAGCACA GAGATATATA GCAAAGAGAT 5461 ACTTTTGAGC AATGTTTGTG GAAGCGGTAT TCGCAATGGG AAGCTCCACC CCGGTTGATA 5521 ATCAGAAAAG CCCCAAAAAC AGGAAGATTG TATAAGCAAA TATTTAAATT GTAAACGTTA 5581 ATATTTTGTT AAAATTCGCG TTAAATTTTT GTTAAATCAG CTCATTTTTT AACGAATAGC 5641 CCGAAATCGG CAAAATCCCT TATAAATCAA AAGAATAGAC CGAGATAGGG TTGAGTGTTG 5701 TTCCAGTTTC CAACAAGAGT CCACTATTAA AGAACGTGGA CTCCAACGTC AAAGGGCGAA 5761 AAAGGGTCTA TCAGGGCGAT GGCCCACTAC GTGAACCATC ACCCTAATCA AGTTTTTTGG 5821 GGTCGAGGTG CCGTAAAGCA GTAAATCGGA AGGGTAAACG GATGCCCCCA TTTAGAGCTT 5881 GACGGGGAAA GCCGGCGAAC GTGGCGAGAA AGGAAGGGAA GAAAGCGAAA GGAGCGGGGG 5941 CTAGGGCGGT GGGAAGTGTA GGGGTCACGC TGGGCGTAAC CACCACACCC GCCGCGCTTA 6001 ATGGGGCGCT ACAGGGCGCG TGGGGATGAT CCACTAGT //
pYES2-NTA其他酵母表达载体：
