北京华越洋生物
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首页    载体及质粒    pYES2/NT A
pYES2NTA

pYES2/NT A

pYES2/NT A

pYES2/NT A



 

编号

载体名称

北京华越洋VECT76333

pYES2/NT A

 

pYES2/NT A载体基本信息:

质粒类型:

酿酒酵母表达载体

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:

Ampicillin (氨苄青霉素)

筛选标记:

URA3

备注:

利用半乳糖诱导蛋白在酿酒酵母中表达。


pYES2/NT A载体图谱:

pYES2/NT A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。

7. 在酵母中,利用载体上的2μ复制子维持质粒的高拷贝。


pYES2/NT A载体序列:

 

ACGGATTAGAAGCCGCCGAGCGGGTGACAGCCCTCCGAAGGAAGACTCTCCTCCGTGCGTCCTCGTCTTC
ACCGGTCGCGTTCCTGAAACGCAGATGTGCCTCGCGCCGCACTGCTCCGAACAATAAAGATTCTACAATA
CTAGCTTTTATGGTTATGAAGAGGAAAAATTGGCAGTAACCTGGCCCCACAAACCTTCAAATGAACGAAT
CAAATTAACAACCATAGGATGATAATGCGATTAGTTTTTTAGCCTTATTTCTGGGGTAATTAATCAGCGA
AGCGATGATTTTTGATCTATTAACAGATATATAAATGCAAAAACTGCATAACCACTTTAACTAATACTTT
CAACATTTTCGGTTTGTATTACTTCTTATTCAAATGTAATAAAAGTATCAACAAAAAATTGTTAATATAC
CTCTATACTTTAACGTCAAGGAGAAAAAACCCCGGATCGGACTACTAGCAGCTGTAATACGACTCACTAT
AGGGAATATTAAGCTTACCATGGGGGGTTCTCATCATCATCATCATCATGGTATGGCTAGCATGACTGGT
GGACAGCAAATGGGTCGGGATCTGTACGACGATGACGATAAGGTACCTAGGATCCAGTGTGGTGGAATTC
TGCAGATATCCAGCACAGTGGCGGCCGCTCGAGTCTAGAGGGCCCTTCGAAGGTAAGCCTATCCCTAACC
CTCTCCTCGGTCTCGATTCTACGCGTACCGGTCATCATCACCATCACCATTGAGTTTAAACCCGCTGATC
CTAGAGGGCCGCATCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCCCCCACATCCGCTCTAA
CCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATT
AAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATA
CTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGCAAGCTGCGGCCCTGCATTA
ATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGAC
TCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCA
CAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGCCCAGGAACCGTAAAAA
GGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGT
CAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCT
CTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTC
TCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAA
CCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACG
ACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGA
GTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAG
CCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTT
TTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTAC
GGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATC
TTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGT
CTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGT
TGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGCGCTTACCATCTGGCCCCAGTGCTGCAATG
ATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGC
GCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAG
TAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGGCATTGCTACAGGCATCGTGGTGTCACTCTCGTCG
TTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCA
AAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCAT
GGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAG
TACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGG
ATAATAGTGTATCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACT
CTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCA
TCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAA
GGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATGGGTAATAACTGATATAATTAAATT
GAAGCTCTAATTTGTGAGTTTAGTATACATGCATTTACTTATAATACAGTTTTTTAGTTTTGCTGGCCGC
ATCTTCTCAAATATGCTTCCCAGCCTGCTTTTCTGTAACGTTCACCCTCTACCTTAGCATCCCTTCCCTT
TGCAAATAGTCCTCTTCCAACAATAATAATGTCAGATCCTGTAGAGACCACATCATCCACGGTTCTATAC
TGTTGACCCAATGCGTCTCCCTTGTCATCTAAACCCACACCGGGTGTCATAATCAACCAATCGTAACCTT
CATCTCTTCCACCCATGTCTCTTTGAGCAATAAAGCCGATAACAAAATCTTTGTCGCTCTTCGCAATGTC
AACAGTACCCTTAGTATATTCTCCAGTAGATAGGGAGCCCTTGCATGACAATTCTGCTAACATCAAAAGG
CCTCTAGGTTCCTTTGTTACTTCTTCTGCCGCCTGCTTCAAACCGCTAACAATACCTGGGCCCACCACAC
CGTGTGCATTCGTAATGTCTGCCCATTCTGCTATTCTGTATACACCCGCAGAGTACTGCAATTTGACTGT
ATTACCAATGTCAGCAAATTTTCTGTCTTCGAAGAGTAAAAAATTGTACTTGGCGGATAATGCCTTTAGC
GGCTTAACTGTGCCCTCCATGGAAAAATCAGTCAAGATATCCACATGTGTTTTTAGTAAACAAATTTTGG
GACCTAATGCTTCAACTAACTCCAGTAATTCCTTGGTGGTACGAACATCCAATGAAGCACACAAGTTTGT
TTGCTTTTCGTGCATGATATTAAATAGCTTGGCAGCAACAGGACTAGGATGAGTAGCAGCACGTTCCTTA
TATGTAGCTTTCGACATGATTTATCTTCGTTTCCTGCAGGTTTTTGTTCTGTGCAGTTGGGTTAAGAATA
CTGGGCAATTTCATGTTTCTTCAACACTACATATGCGTATATATACCAATCTAAGTCTGTGCTCCTTCCT
TCGTTCTTCCTTCTGTTCGGAGATTACCGAATCAAAAAAATTTCAAAGAAACCGAAATCAAAAAAAAGAA
TAAAAAAAAAATGATGAATTGAATTGAAAAGCTAGCTTATCGATGATAAGCTGTCAAAGATGAGAATTAA
TTCCACGGACTATAGACTATACTAGATACTCCGTCTACTGTACGATACACTTCCGCTCAGGTCCTTGTCC
TTTAACGAGGCCTTACCACTCTTTTGTTACTCTATTGATCCAGCTCAGCAAAGGCAGTGTGATCTAAGAT
TCTATCTTCGCGATGTAGTAAAACTAGCTAGACCGAGAAAGAGACTAGAAATGCAAAAGGCACTTCTACA
ATGGCTGCCATCATTATTATCCGATGTGACGCTGCAGCTTCTCAATGATATTCGAATACGCTTTGAGGAG
ATACAGCCTAATATCCGACAAACTGTTTTACAGATTTACGATCGTACTTGTTACCCATCATTGAATTTTG
AACATCCGAACCTGGGAGTTTTCCCTGAAACAGATAGTATATTTGAACCTGTATAATAATATATAGTCTA
GCGCTTTACGGAAGACAATGTATGTATTTCGGTTCCTGGAGAAACTATTGCATCTATTGCATAGGTAATC
TTGCACGTCGCATCCCCGGTTCATTTTCTGCGTTTCCATCTTGCACTTCAATAGCATATCTTTGTTAACG
AAGCATCTGTGCTTCATTTTGTAGAACAAAAATGCAACGCGAGAGCGCTAATTTTTCAAACAAAGAATCT
GAGCTGCATTTTTACAGAACAGAAATGCAACGCGAAAGCGCTATTTTACCAACGAAGAATCTGTGCTTCA
TTTTTGTAAAACAAAAATGCAACGCGACGAGAGCGCTAATTTTTCAAACAAAGAATCTGAGCTGCATTTT
TACAGAACAGAAATGCAACGCGAGAGCGCTATTTTACCAACAAAGAATCTATACTTCTTTTTTGTTCTAC
AAAAATGCATCCCGAGAGCGCTATTTTTCTAACAAAGCATCTTAGATTACTTTTTTTCTCCTTTGTGCGC
TCTATAATGCAGTCTCTTGATAACTTTTTGCACTGTAGGTCCGTTAAGGTTAGAAGAAGGCTACTTTGGT
GTCTATTTTCTCTTCCATAAAAAAAGCCTGACTCCACTTCCCGCGTTTACTGATTACTAGCGAAGCTGCG
GGTGCATTTTTTCAAGATAAAGGCATCCCCGATTATATTCTATACCGATGTGGATTGCGCATACTTTGTG
AACAGAAAGTGATAGCGTTGATGATTCTTCATTGGTCAGAAAATTATGAACGGTTTCTTCTATTTTGTCT
CTATATACTACGTATAGGAAATGTTTACATTTTCGTATTGTTTTCGATTCACTCTATGAATAGTTCTTAC
TACAATTTTTTTGTCTAAAGAGTAATACTAGAGATAAACATAAAAAATGTAGAGGTCGAGTTTAGATGCA
AGTTCAAGGAGCGAAAGGTGGATGGGTAGGTTATATAGGGATATAGCACAGAGATATATAGCAAAGAGAT
ACTTTTGAGCAATGTTTGTGGAAGCGGTATTCGCAATGGGAAGCTCCACCCCGGTTGATAATCAGAAAAG
CCCCAAAAACAGGAAGATTGTATAAGCAAATATTTAAATTGTAAACGTTAATATTTTGTTAAAATTCGCG
TTAAATTTTTGTTAAATCAGCTCATTTTTTAACGAATAGCCCGAAATCGGCAAAATCCCTTATAAATCAA
AAGAATAGACCGAGATAGGGTTGAGTGTTGTTCCAGTTTCCAACAAGAGTCCACTATTAAAGAACGTGGA
CTCCAACGTCAAAGGGCGAAAAAGGGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACCCTAATCA
AGTTTTTTGGGGTCGAGGTGCCGTAAAGCAGTAAATCGGAAGGGTAAACGGATGCCCCCATTTAGAGCTT
GACGGGGAAAGCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAAAGCGAAAGGAGCGGGGGCTAGGGCGGT
GGGAAGTGTAGGGGTCACGCTGGGCGTAACCACCACACCCGCCGCGCTTAATGGGGCGCTACAGGGCGCG
TGGGGATGATCCACTAGT

 

pYES2/NT A载体相关的酵母表达载体:

pKLAC1YIplac211
pHiSi-1YEplac181
pHISiYCplac22
pGBKlampYX212-VHB
pGBK-53pYES2-flag
pGAD424pYES2/NT C
pGAD53mpYES2/NT B
pFLD-cat Z+pYES2/NT A
pFLD-cat A+pYES2/CT/α-Factor
pFLD1 Z+pYADE4
pESP-3pUG35
pESP-2pP-αSUMO3
pESP-1pPICZalpha GB
pDR196pPICZalpha GA
pDNR1pPICZalpha FC
pBLADE-SXpPICZalpha E
pACT2 ADpMETα C
p426 GALpMETα B
P416 GPDpMETα A
pPIC6αApYD1
pRS406p53his
pRS416pCL1
pSEP1pSos
pHisSi-1pGBKT7-53
pMyrpFLDα
pGBKT7-LampADH2
pDisplaypYIP5
pPinkα-HCpDR195
pPIC3.5KpYES2
pAUR123pPIC ZαD
pPICZ CYcplac33
pHIC-PIpYIP211
pRS426galpYES2-kan
p416GFDpPICZα B
pYCP211pESC-Leu
pRS426pYES2-NTC
pESC-TRPpMETαB
pYES3/CTpMET A
pMETαCpHIL-S1
pMET BpPICZα C
pHIL-D2pPIC3.5
pGAPZα ApUG66
pAO815pDEST32
pGAPZ CpYC2/NT A
pYC2/NT CpYES-DEST52
pYES2/CTpPIC6αC
pPIC6 ApRS403
pGAG424pSos-MAFB
pGADT7-TpBridge
pHis2pACT2-AD
pGBKT7pFLD/CAT
pFLDpYES2-EGFP
Ycp22lac-EGFPpPIC9k-His
SUMO proteasepPICZ A
pGADT7LA503
BFD15pPIC ZαFC
pPIC ZαGBpYEPlac112
pYEPlac195pYRP7
pYX212pFA6a-FGP(S65T)-kanMX6
pRS41HpESC-URA
pRS316pYES6/CT
pESC-HispYES2-NTB
pYES2-NTApMET C
pMETαApPink-HC
pPink-LCpGAPZ B
pGAPZα CpPICZα A
pPICZ BpYC2/CT
pPIC9pRS405
pGAPZ ApRS414
pAD-GAL4-2.1pSEP2
pYC2/NT Bp53blue
pPIC6 CpRS415
pPIC6αBpSEP3
pRS413pBD-GAL4-Cam