pYES2-NTC

pYES2-NTC

载体基本信息

出品公司:	Ybscience
载体名称:	pYES2-NTC, pYES2/NT C
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:	6037 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

载体质粒图谱和多克隆位点信息

载体简介

pYES2 NTC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。

载体序列

LOCUS       pYES2/NT C	6037 bp 	DNA     circular	SYN
DEFINITION  pYES2/NT C
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     VNTAUTHORNAME|biofeng.com|
FEATURES             Location/Qualifiers
     source          1..6037
                     /organism="pYES2/NT C"
                     /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      790..1035
                     /label="CYC1_terminator"
     misc_feature    complement(799..817)
                     /label="CYC1_primer"
     misc_feature    complement(799..817)
                     /label="pYESTrp_rev_primer"
     rep_origin      complement(1263..1882)
                     /label="pBR322_origin"
     CDS             complement(2037..2897)
                     /label="ORF frame 3"
     CDS             complement(2993..3796)
                     /label="ORF frame 1"
     gene            complement(2996..3796)
                     /label="URA3"
                     /gene="URA3" 
     CDS             3285..3761
                     /label="ORF frame 3"
     promoter        complement(3798..4023)
                     /label="URA3_promoter"
     rep_origin      4026..5497
                     /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 AGGTACCCGG ATCCAGTGTG GTGGAATTCT GCAGATATCC AGCACAGTGG CGGCCGCTCG
  661 AGTCTAGAGG GCCCTTCGAA GGTAAGCCTA TCCCTAACCC TCTCCTCGGT CTCGATTCTA
  721 CGCGTACCGG TCATCATCAC CATCACCATT GAGTTTAAAC CCGCTGATCC TAGAGGGCCG
  781 CATCATGTAA TTAGTTATGT CACGCTTACA TTCACGCCCT CCCCCCACAT CCGCTCTAAC
  841 CGAAAAGGAA GGAGTTAGAC AACCTGAAGT CTAGGTCCCT ATTTATTTTT TTATAGTTAT
  901 GTTAGTATTA AGAACGTTAT TTATATTTCA AATTTTTCTT TTTTTTCTGT ACAGACGCGT
  961 GTACGCATGT AACATTATAC TGAAAACCTT GCTTGAGAAG GTTTTGGGAC GCTCGAAGGC
 1021 TTTAATTTGC AAGCTGCGGC CCTGCATTAA TGAATCGGCC AACGCGCGGG GAGAGGCGGT
 1081 TTGCGTATTG GGCGCTCTTC CGCTTCCTCG CTCACTGACT CGCTGCGCTC GGTCGTTCGG
 1141 CTGCGGCGAG CGGTATCAGC TCACTCAAAG GCGGTAATAC GGTTATCCAC AGAATCAGGG
 1201 GATAACGCAG GAAAGAACAT GTGAGCAAAA GGCCAGCAAA AGCCCAGGAA CCGTAAAAAG
 1261 GCCGCGTTGC TGGCGTTTTT CCATAGGCTC CGCCCCCCTG ACGAGCATCA CAAAAATCGA
 1321 CGCTCAAGTC AGAGGTGGCG AAACCCGACA GGACTATAAA GATACCAGGC GTTTCCCCCT
 1381 GGAAGCTCCC TCGTGCGCTC TCCTGTTCCG ACCCTGCCGC TTACCGGATA CCTGTCCGCC
 1441 TTTCTCCCTT CGGGAAGCGT GGCGCTTTCT CATAGCTCAC GCTGTAGGTA TCTCAGTTCG
 1501 GTGTAGGTCG TTCGCTCCAA GCTGGGCTGT GTGCACGAAC CCCCCGTTCA GCCCGACCGC
 1561 TGCGCCTTAT CCGGTAACTA TCGTCTTGAG TCCAACCCGG TAAGACACGA CTTATCGCCA
 1621 CTGGCAGCAG CCACTGGTAA CAGGATTAGC AGAGCGAGGT ATGTAGGCGG TGCTACAGAG
 1681 TTCTTGAAGT GGTGGCCTAA CTACGGCTAC ACTAGAAGGA CAGTATTTGG TATCTGCGCT
 1741 CTGCTGAAGC CAGTTACCTT CGGAAAAAGA GTTGGTAGCT CTTGATCCGG CAAACAAACC
 1801 ACCGCTGGTA GCGGTGGTTT TTTTGTTTGC AAGCAGCAGA TTACGCGCAG AAAAAAAGGA
 1861 TCTCAAGAAG ATCCTTTGAT CTTTTCTACG GGGTCTGACG CTCAGTGGAA CGAAAACTCA
 1921 CGTTAAGGGA TTTTGGTCAT GAGATTATCA AAAAGGATCT TCACCTAGAT CCTTTTAAAT
 1981 TAAAAATGAA GTTTTAAATC AATCTAAAGT ATATATGAGT AAACTTGGTC TGACAGTTAC
 2041 CAATGCTTAA TCAGTGAGGC ACCTATCTCA GCGATCTGTC TATTTCGTTC ATCCATAGTT
 2101 GCCTGACTCC CCGTCGTGTA GATAACTACG ATACGGGAGC GCTTACCATC TGGCCCCAGT
 2161 GCTGCAATGA TACCGCGAGA CCCACGCTCA CCGGCTCCAG ATTTATCAGC AATAAACCAG
 2221 CCAGCCGGAA GGGCCGAGCG CAGAAGTGGT CCTGCAACTT TATCCGCCTC CATCCAGTCT
 2281 ATTAATTGTT GCCGGGAAGC TAGAGTAAGT AGTTCGCCAG TTAATAGTTT GCGCAACGTT
 2341 GTTGGCATTG CTACAGGCAT CGTGGTGTCA CTCTCGTCGT TTGGTATGGC TTCATTCAGC
 2401 TCCGGTTCCC AACGATCAAG GCGAGTTACA TGATCCCCCA TGTTGTGCAA AAAAGCGGTT
 2461 AGCTCCTTCG GTCCTCCGAT CGTTGTCAGA AGTAAGTTGG CCGCAGTGTT ATCACTCATG
 2521 GTTATGGCAG CACTGCATAA TTCTCTTACT GTCATGCCAT CCGTAAGATG CTTTTCTGTG
 2581 ACTGGTGAGT ACTCAACCAA GTCATTCTGA GAATAGTGTA TGCGGCGACC GAGTTGCTCT
 2641 TGCCCGGCGT CAATACGGGA TAATAGTGTA TCACATAGCA GAACTTTAAA AGTGCTCATC
 2701 ATTGGAAAAC GTTCTTCGGG GCGAAAACTC TCAAGGATCT TACCGCTGTT GAGATCCAGT
 2761 TCGATGTAAC CCACTCGTGC ACCCAACTGA TCTTCAGCAT CTTTTACTTT CACCAGCGTT
 2821 TCTGGGTGAG CAAAAACAGG AAGGCAAAAT GCCGCAAAAA AGGGAATAAG GGCGACACGG
 2881 AAATGTTGAA TACTCATACT CTTCCTTTTT CAATGGGTAA TAACTGATAT AATTAAATTG
 2941 AAGCTCTAAT TTGTGAGTTT AGTATACATG CATTTACTTA TAATACAGTT TTTTAGTTTT
 3001 GCTGGCCGCA TCTTCTCAAA TATGCTTCCC AGCCTGCTTT TCTGTAACGT TCACCCTCTA
 3061 CCTTAGCATC CCTTCCCTTT GCAAATAGTC CTCTTCCAAC AATAATAATG TCAGATCCTG
 3121 TAGAGACCAC ATCATCCACG GTTCTATACT GTTGACCCAA TGCGTCTCCC TTGTCATCTA
 3181 AACCCACACC GGGTGTCATA ATCAACCAAT CGTAACCTTC ATCTCTTCCA CCCATGTCTC
 3241 TTTGAGCAAT AAAGCCGATA ACAAAATCTT TGTCGCTCTT CGCAATGTCA ACAGTACCCT
 3301 TAGTATATTC TCCAGTAGAT AGGGAGCCCT TGCATGACAA TTCTGCTAAC ATCAAAAGGC
 3361 CTCTAGGTTC CTTTGTTACT TCTTCTGCCG CCTGCTTCAA ACCGCTAACA ATACCTGGGC
 3421 CCACCACACC GTGTGCATTC GTAATGTCTG CCCATTCTGC TATTCTGTAT ACACCCGCAG
 3481 AGTACTGCAA TTTGACTGTA TTACCAATGT CAGCAAATTT TCTGTCTTCG AAGAGTAAAA
 3541 AATTGTACTT GGCGGATAAT GCCTTTAGCG GCTTAACTGT GCCCTCCATG GAAAAATCAG
 3601 TCAAGATATC CACATGTGTT TTTAGTAAAC AAATTTTGGG ACCTAATGCT TCAACTAACT
 3661 CCAGTAATTC CTTGGTGGTA CGAACATCCA ATGAAGCACA CAAGTTTGTT TGCTTTTCGT
 3721 GCATGATATT AAATAGCTTG GCAGCAACAG GACTAGGATG AGTAGCAGCA CGTTCCTTAT
 3781 ATGTAGCTTT CGACATGATT TATCTTCGTT TCCTGCAGGT TTTTGTTCTG TGCAGTTGGG
 3841 TTAAGAATAC TGGGCAATTT CATGTTTCTT CAACACTACA TATGCGTATA TATACCAATC
 3901 TAAGTCTGTG CTCCTTCCTT CGTTCTTCCT TCTGTTCGGA GATTACCGAA TCAAAAAAAT
 3961 TTCAAAGAAA CCGAAATCAA AAAAAAGAAT AAAAAAAAAA TGATGAATTG AATTGAAAAG
 4021 CTAGCTTATC GATGATAAGC TGTCAAAGAT GAGAATTAAT TCCACGGACT ATAGACTATA
 4081 CTAGATACTC CGTCTACTGT ACGATACACT TCCGCTCAGG TCCTTGTCCT TTAACGAGGC
 4141 CTTACCACTC TTTTGTTACT CTATTGATCC AGCTCAGCAA AGGCAGTGTG ATCTAAGATT
 4201 CTATCTTCGC GATGTAGTAA AACTAGCTAG ACCGAGAAAG AGACTAGAAA TGCAAAAGGC
 4261 ACTTCTACAA TGGCTGCCAT CATTATTATC CGATGTGACG CTGCAGCTTC TCAATGATAT
 4321 TCGAATACGC TTTGAGGAGA TACAGCCTAA TATCCGACAA ACTGTTTTAC AGATTTACGA
 4381 TCGTACTTGT TACCCATCAT TGAATTTTGA ACATCCGAAC CTGGGAGTTT TCCCTGAAAC
 4441 AGATAGTATA TTTGAACCTG TATAATAATA TATAGTCTAG CGCTTTACGG AAGACAATGT
 4501 ATGTATTTCG GTTCCTGGAG AAACTATTGC ATCTATTGCA TAGGTAATCT TGCACGTCGC
 4561 ATCCCCGGTT CATTTTCTGC GTTTCCATCT TGCACTTCAA TAGCATATCT TTGTTAACGA
 4621 AGCATCTGTG CTTCATTTTG TAGAACAAAA ATGCAACGCG AGAGCGCTAA TTTTTCAAAC
 4681 AAAGAATCTG AGCTGCATTT TTACAGAACA GAAATGCAAC GCGAAAGCGC TATTTTACCA
 4741 ACGAAGAATC TGTGCTTCAT TTTTGTAAAA CAAAAATGCA ACGCGACGAG AGCGCTAATT
 4801 TTTCAAACAA AGAATCTGAG CTGCATTTTT ACAGAACAGA AATGCAACGC GAGAGCGCTA
 4861 TTTTACCAAC AAAGAATCTA TACTTCTTTT TTGTTCTACA AAAATGCATC CCGAGAGCGC
 4921 TATTTTTCTA ACAAAGCATC TTAGATTACT TTTTTTCTCC TTTGTGCGCT CTATAATGCA
 4981 GTCTCTTGAT AACTTTTTGC ACTGTAGGTC CGTTAAGGTT AGAAGAAGGC TACTTTGGTG
 5041 TCTATTTTCT CTTCCATAAA AAAAGCCTGA CTCCACTTCC CGCGTTTACT GATTACTAGC
 5101 GAAGCTGCGG GTGCATTTTT TCAAGATAAA GGCATCCCCG ATTATATTCT ATACCGATGT
 5161 GGATTGCGCA TACTTTGTGA ACAGAAAGTG ATAGCGTTGA TGATTCTTCA TTGGTCAGAA
 5221 AATTATGAAC GGTTTCTTCT ATTTTGTCTC TATATACTAC GTATAGGAAA TGTTTACATT
 5281 TTCGTATTGT TTTCGATTCA CTCTATGAAT AGTTCTTACT ACAATTTTTT TGTCTAAAGA
 5341 GTAATACTAG AGATAAACAT AAAAAATGTA GAGGTCGAGT TTAGATGCAA GTTCAAGGAG
 5401 CGAAAGGTGG ATGGGTAGGT TATATAGGGA TATAGCACAG AGATATATAG CAAAGAGATA
 5461 CTTTTGAGCA ATGTTTGTGG AAGCGGTATT CGCAATGGGA AGCTCCACCC CGGTTGATAA
 5521 TCAGAAAAGC CCCAAAAACA GGAAGATTGT ATAAGCAAAT ATTTAAATTG TAAACGTTAA
 5581 TATTTTGTTA AAATTCGCGT TAAATTTTTG TTAAATCAGC TCATTTTTTA ACGAATAGCC
 5641 CGAAATCGGC AAAATCCCTT ATAAATCAAA AGAATAGACC GAGATAGGGT TGAGTGTTGT
 5701 TCCAGTTTCC AACAAGAGTC CACTATTAAA GAACGTGGAC TCCAACGTCA AAGGGCGAAA
 5761 AAGGGTCTAT CAGGGCGATG GCCCACTACG TGAACCATCA CCCTAATCAA GTTTTTTGGG
 5821 GTCGAGGTGC CGTAAAGCAG TAAATCGGAA GGGTAAACGG ATGCCCCCAT TTAGAGCTTG
 5881 ACGGGGAAAG CCGGCGAACG TGGCGAGAAA GGAAGGGAAG AAAGCGAAAG GAGCGGGGGC
 5941 TAGGGCGGTG GGAAGTGTAG GGGTCACGCT GGGCGTAACC ACCACACCCG CCGCGCTTAA
 6001 TGGGGCGCTA CAGGGCGCGT GGGGATGATC CACTAGT