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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃
- 保质期：
6个月
- 英文名：
pMET A
- 库存：
大量
- 供应商：
钰博生物
- 规格：
1ug/5ug
pMET A
pMETA载体基本信息
| 出品公司: | Ybscience |
|---|---|
| 载体名称: | pMET A, pMETA |
| 质粒类型: | 毕赤酵母蛋白表达载体（methanoloica酵母） |
| 表达水平: | 高拷贝 |
| 诱导方法: | 甲醇诱导 |
| 启动子: | P-AUG1 |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 7778 bp |
| 5' 测序引物及序列: | AUG1-F: 5´-CAATTTACATCTTTATTTATTAACG-3´ |
| 3' 测序引物及序列: | AUG1-R: 5´-GAAGAGAAAAACATTAGTTGGC-3´ |
| 载体标签: | C-V5， C-His |
| 载体抗性: | 氨苄 |
| 筛选标记: | ADE2 |
| 备注: | 利用甲醇诱导蛋白在methanoloica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 |
pMETA载体质粒图谱和多克隆位点信息
pMETA载体简介
pMETA载体序列
LOCUS pMET A 7778 bp DNA circular SYN
DEFINITION pMET A
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..7778
/organism="pMET A"
/mol_type="other DNA"
misc_feature 1106..1130
/label="AUG1_fwd_primer"
misc_feature complement(1397..1418)
/label="AUG1_rev_primer"
CDS 2820..3395
/label="ORF frame 3"
CDS 3401..4534
/label="ORF frame 2"
CDS 4698..5696
/label="ORF frame 3"
gene complement(6509..7369)
/label="Ampicillin"
/gene="Ampicillin"
CDS complement(6509..7369)
/label="ORF frame 2"
promoter complement(7411..7439)
/label="AmpR_promoter"
ORIGIN
1 GGCGCGCCTT AATTAAGGCC GGCCCCTGCA GGGGGCCCCG AGATGGTACA TACTTAAAAG
61 CTGCCATATT GAGGAACTTC AAAGTTTTAT CTGTTTTTAG AATTAAAAGA CGATTGTTGT
121 AACAAAACGT TGTGCCTACA TAAACTCAAA TTAATGGAAA TAGCCTGTTT TGAAAAATAC
181 ACCTTCTTAA GTACTGACAA AGTTTTGTTA AATGACTATC GAACAAGCCA TGAAATAGCA
241 CATTTCTGCC AGTCACTTTT AACACTTTCC TGCTTGCTGG TTGACTCTCC TCATACAAAC
301 ACCCAAAAGG GAAACTTTCA GTGTGGGGAC ACTTGACATC TCACATGCAC CCCAGATTAA
361 TTTCCCCAGA CGATGCGGAG ACAAGACAAA ACAACCCTTT GTCCTGCTCT TTTCTTTCTC
421 ACACCGCGTG GGTGTGTGCG CAGGCAGGCA GGCAGGCAGC GGGCTGCCTG CCATCTCTAA
481 TCGCTGCTCC TCCCCCCTGG CTTCAAATAA CAGCCTGCTG CTATCTGTGA CCAGATTGGG
541 ACACCCCCCT CCCCTCCGAA TGATCCATCA CCTTTTGTCG TACTCCGACA ATGATCCTTC
601 CCTGTCATCT TCTGGCAATC AGCTCCTTCA ATAATTAAAT CAAATAAGCA TAAATAGTAA
661 AATCGCATAC AAACGTCATG AAAAGTTTTA TCTCTATGGC CAACGGATAG TCTATCTGCT
721 TAATTCCATC CACTTTGGGA ACCGTTCTCT CTTTACCCCA GATTCTCAAA GCTAATATCT
781 GCCCCTTGTC TATTGTCCTT TCTCCGTGTA CAAGCGGAGC TTTTGCCTCC CATCCTCTTG
841 CTTTGTTTCG GTTATTTTTT TTTTCTTTTG AAACTCTTGG TCAAATCAAA TCAAACAAAA
901 CCAAACCTTC TATTCCATCA GATCAACCTT GTTCAACATT CTATAAATCG ATATAAATAT
961 AACCTTATCC CTCCCTTGTT TTTTACCAAT TAATCAATCT TCAAATTTCA AATATTTTCT
1021 ACTTGCTTTA TTACTCAGTA TTAACATTTG TTTAAACCAA CTATAACTTT TAACTGGCTT
1081 TAGAAGTTTT ATTTAACATC AGTTTCAATT TACATCTTTA TTTATTAACG AAATCTTTAC
1141 GAATTAACTC AATCAAAACT TTTACGAAAA AAAAATCTTA CTATTAAGAG CTCGAGAGGA
1201 ATTCACGTGG CCCAGCCGGC CGTGGATCCA CGCGTCGTCG ACCCGCGGCG GCCGCCAGCT
1261 TCCTAGGGGT AAGCCTATCC CTAACCCTCT CCTCGGTCTC GATTCTACGC GTACCGGTCA
1321 TCATCACCAT CACCATTGAA CTAGTATACA ATTCTAGGGC TGCCTGTTTG GATATTTTTA
1381 TAATTTTTGA GAGTTTGCCA ACTAATGTTT TTCTCTTCTA TGATATTTAT CATGTAGTTG
1441 GATATCTTTT TTTCTTTGAA AAGTTTTTAT TTAGTTCTTT CTCTCTTTTT TATAATGTTT
1501 TCAGTTTCTT TCCTTTATAT TGAAAATACG ATGTTCTTTG TCTTTCTTTT TTAAATCCTC
1561 AATTATAATG TGTACCGTTT GATCAACTTG TAAAATTGTT TGTAGTCCTG TAATCTTAAT
1621 TGACGATTGG TTGATAGTTT ACCCCAGTAT ATTGTGCGGC TGAGATATTA CCCGATTCGG
1681 AAGATTGATT TTTTCAACAG ACATAAACCC ATAGGAGATT TAGTACAGCG ACATATGCTA
1741 ATTGCCTGAT AATGCTCACT AACATTTTCT CCTTTTAATC TGGTATATGC ATACAGGTAT
1801 GTTGATATGG CCTTGTTGAT CAAAGCTACG AAAGTGTAGA CTATTTCCCA AACAGCCCCC
1861 AACCACCACC TCACACGCGC ACACTGCGAA GTGAAAGCGC TCCCTTATTT TCTTCTCCGC
1921 TGGTTCAAAA CAAAAAAGAA TCTAATCTGT AGACACACGC CCCGTAAAAC CCAATCTTCA
1981 TCATCGTAAT TTTCAGTGAC GGACGATCGA TCAACCATAT AGTTTGTTAG TGACTGAGCT
2041 TCAGTAGTTG ACCTCTTGAA GAAATAACGT CTTGAACAAC ATACTTTATA CCTACCAACA
2101 TTCGCCATCA ACAACAGTGT GACACTGACA TAACTTTACA TCCATTCATC GACAAAAAAA
2161 TGTCTCTAGT TCGGATTCAT GCTTATGGGT TAGCTATTTC GCCCAATGTG TCCATCTGAC
2221 ATTACTATTT TGCATTTTAA TTTAATTAGA ACTTGACTAG CGCACTACCA GCATATCATC
2281 TCATTTCCGT AAATACCAAA TGTATTATAT ATTGAAAGCT TTTGACCAGG TTATTATAAA
2341 AGAAACTTCA TGCTCGAAAA AGATCATTTC GAAAAGTTGC CTAGTTTCAT GAAATTTTAA
2401 AGCAGTTTAT ATAAATTTTA CCTTTTGATG CGGAATTGAC TTTTTCTTGA ATAATACATA
2461 ACTTTTCTTA AAAGAATCAA AGACAGATAA AATTTAAGAG ATATTAAATA TTAGTGAGAA
2521 GCCGAGAATT TTGTAACACC AACATAACAC TGACATCTTT AACAACTTTT AATTATGATA
2581 CATTTCTTAC GTCATGATTG ATTATTACAG CTATGCTGAC AAATGACTCT TGTTGCATGG
2641 CTACGAACCG GGTAATACTA AGTGATTGAC TCTTGCTGAC CTTTTATTAA GAACTAAATG
2701 GACAATATTA TGGAGCATTT CATGTATAAA TTGGTGCGTA AAATCGTTGG ATCTCTCTTC
2761 TAAGTACATC CTACTATAAC AATCAAGAAA AACAAGAAAA TCGGACAAAA CAATCAAGTA
2821 TGGATTCTAG AACAGTTGGT ATATTAGGAG GGGGACAATT GGGACGTATG ATTGTTGAGG
2881 CAGCAAACAG GCTCAACATT AAGACGGTAA TACTAGATGC TGAAAATTCT CCTGCCAAAC
2941 AAATAAGCAA CTCCAATGAC CACGTTAATG GCTCCTTTTC CAATCCTCTT GATATCGAAA
3001 AACTAGCTGA AAAATGTGAT GTGCTAACGA TTGAGATTGA GCATGTTGAT GTTCCTACAC
3061 TAAAGAATCT TCAAGTAAAA CATCCCAAAT TAAAAATTTA CCCTTCTCCA GAAACAATCA
3121 GATTGATACA AGACAAATAT ATTCAAAAAG AGCATTTAAT CAAAAATGGT ATAGCAGTTA
3181 CCCAAAGTGT TCCTGTGGAA CAAGCCAGTG AGACGTCCCT ATTGAATGTT GGAAGAGATT
3241 TGGGTTTTCC ATTCGTCTTG AAGTCGAGGA CTTTGGCATA CGATGGAAGA GGTAACTTCG
3301 TTGTAAAGAA TAAGGAAATG ATTCCGGAAG CTTTGGAAGT ACTGAAGGAT CGTCCTTTGT
3361 CGCCGAAAAA TGGGCACCAT TTACTAAAGA ATTAGCAGTC ATGATTGTGA GATCTGTTAA
3421 CGGTTTAGTG TTTTCTTACC CAATTGTAGA GACTATCCAC AAGGACAATA TTTGTGACTT
3481 ATGTTATGCG CCTGCTAGAG TTCCGGACTC CGTTCAACTT AAGGCGAAGT TGTTGGCAGA
3541 AAATGCAATC AAATCTTTTC CCGGTTGTGG TATATTTGGT GTGGAAATGT TCTATTTAGA
3601 AACAGGGGAA TTGCTTATTA ACGAAATTGC CCCAAGGCCT CACAACTCTG GACATTATAC
3661 CATTGATGCT TGCGTCACTT CTCAATTTGA AGCTCATTTG AGATCAATAT TGGATTTGCC
3721 AATGCCAAAG AATTTCACAT CTTTCTCCAC CATTACAACG AACGCCATTA TGCTAAATGT
3781 TCTTGGAGAC AAACATACAA AAGATAAAGA GCTAGAAACT TGCGAAAGAG CATTGGCGAC
3841 TCCAGGTTCC TCAGTGTACT TATATGGAAA AGAGTCTAGA CCTAACAGAA AAGTAGGTCA
3901 CATAAATATT ATTGCCTCCA GTATGGCGGA ATGTGAACAA AGGCTGAACT ACATTACAGG
3961 TAGAACTGAT ATTCCAATCA AAATCTCTGT CGCTCAAAAG TTGGACTTGG AAGCAATGGT
4021 CAAACCATTG GTTGGAATCA TCATGGGATC AGACTCTGAC TTGCCGGTAA TGTCTGCCGC
4081 ATGTGCGGTT TTAAAAGATT TTGGCGTTCC ATTTGAAGTG ACAATAGTCT CTGCTCATAG
4141 AACTCCACAT AGGATGTCAG CATATGCTAT TTCCGCAAGC AAGCGTGGAA TTAAAACAAT
4201 TATCGCTGGA GCTGGTGGGG CTGCTCACTT GCCAGGTATG GTGGCTGCAA TGACACCACT
4261 TCCTGTCATC GGTGTGCCCG TAAAAGGTTC TTGTCTAGAT GGAGTAGATT CTTTACATTC
4321 AATTGTGCAA ATGCCTAGAG GTGTTCCAGT AGCTACCGTC GCTATTAATA ATAGTACGAA
4381 CGCTGCGCTG TTGGCTGTCA GACTGCTTGG CGCTTATGAT TCAAGTTATA CAACGAAAAT
4441 GGAACAGTTT TTATTAAAGC AAGAAGAAGA AGTTCTTGTC AAAGCACAAA AGTTAGAAAC
4501 TGTCGGTTAC GAAGCTTATC TAGAAAACAA GTAATATATA AGTTTATTGA TATACTTGTA
4561 CAGCAAATAA TTATAAAATG ATATACCTAT TTTTTAGGCT TTGTTATGAT TACATCAAAT
4621 GTGGACTTCA TACATAGAAA TCAACGCTTA CAGGTGTCCT TTTTTAAGAA TTTCATACAT
4681 AAGATCTACT AGAATCAATG TTAAACCTGT TATCATTAGA ACAGCCATCT CCAGAAGGTT
4741 CTATCCAAGT GAAAACTTTG GTCTTCAAGC CAAAGACTGC TAAAACACAA ACCCCAGTCC
4801 CAATTATTGT TGTTGCTGAA ACTTCTACTT CTACTCCGGG TGGTGTTATA GCCAAAGCTT
4861 CGGGCACTAA AGAGCCAAGA TTAGCAAACG ATGAATTGAC CAACCAATTC TTTGGTTCTC
4921 CTGCTAAAGA GTTTTCTATT GCCAGCTTAT CAAACGAACA CAAAGACGGT AAAGTCAAGA
4981 TCATTATTGA TGCTGGGATC AAAGATGATG ACTCGACTAA TTACGTTATT TCCAACACTG
5041 TTGCTAAGTT AGTTTTAACC GGTAAGCAAT TATTAGGTTT CTTATCCAAC TACGAACCAA
5101 CCATCGTTGA TTTTACTGCT GAAGCTGCAA CCGCTGCCCC AACTCCAGTT GTTAAGAAAG
5161 AACCTGCTGC ATCTGGTAAG CTTGATGATG CCAAATTAAT TGGTGTTACT GCTGACAAGG
5221 AAAAAGATTT CTCCAACTGG TACTCCCAAA TTCTAATGAA GGGTGAAATG TTAGACTACT
5281 ACGATGTCTC TGGTTGTTAC ATTATGAGAC CAGCTTCTTA CACAATTTGG GAAAAGATTC
5341 AATCTTGGTT CGATTCGAAG ATTAAAGAAG ATGGTGTTAC TAATGCTTAT TTCCCAATGT
5401 TTGTCTCATC CAAGGTTTTA GAAAAGGAAA AGGACCACAT CGATGGTTTC GCTCCTGAAG
5461 TTGCTTGGGT TACCAGAGCT GGTAAATCTG AATTAGAAGA ACCAATTGCT ATTAGACCAA
5521 CTTCTGAAAC TGTTATGGGT ACCCTGCAGG GGCCGGCCTT AATTAAGGCG CGCCCTTCCT
5581 CGCTCACTGA CTCGCTGCGC TCGGTCGTTC GGCTGCGGCG AGCGGTATCA GCTCACTCAA
5641 AGGCGGTAAT ACGGTTATCC ACAGAATCAG GGGATAACGC AGGAAAGAAC ATGTGAGCAA
5701 AAGGCCAGCA AAAGGCCAGG AACCGTAAAA AGGCCGCGTT GCTGGCGTTT TTCCATAGGC
5761 TCCGCCCCCC TGACGAGCAT CACAAAAATC GACGCTCAAG TCAGAGGTGG CGAAACCCGA
5821 CAGGACTATA AAGATACCAG GCGTTTCCCC CTGGAAGCTC CCTCGTGCGC TCTCCTGTTC
5881 CGACCCTGCC GCTTACCGGA TACCTGTCCG CCTTTCTCCC TTCGGGAAGC GTGGCGCTTT
5941 CTCATAGCTC ACGCTGTAGG TATCTCAGTT CGGTGTAGGT CGTTCGCTCC AAGCTGGGCT
6001 GTGTGCACGA ACCCCCCGTT CAGCCCGACC GCTGCGCCTT ATCCGGTAAC TATCGTCTTG
6061 AGTCCAACCC GGTAAGACAC GACTTATCGC CACTGGCAGC AGCCACTGGT AACAGGATTA
6121 GCAGAGCGAG GTATGTAGGC GGTGCTACAG AGTTCTTGAA GTGGTGGCCT AACTACGGCT
6181 ACACTAGAAG AACAGTATTT GGTATCTGCG CTCTGCTGAA GCCAGTTACC TTCGGAAAAA
6241 GAGTTGGTAG CTCTTGATCC GGCAAACAAA CCACCGCTGG TAGCGGTGGT TTTTTTGTTT
6301 GCAAGCAGCA GATTACGCGC AGAAAAAAAG GATCTCAAGA AGATCCTTTG ATCTTTTCTA
6361 CGGGGTCTGA CGCTCAGTGG AACGAAAACT CACGTTAAGG GATTTTGGTC ATGAGATTAT
6421 CAAAAAGGAT CTTCACCTAG ATCCTTTTAA ATTAAAAATG AAGTTTTAAA TCAATCTAAA
6481 GTATATATGA GTAAACTTGG TCTGACAGTT ACCAATGCTT AATCAGTGAG GCACCTATCT
6541 CAGCGATCTG TCTATTTCGT TCATCCATAG TTGCCTGACT CCCCGTCGTG TAGATAACTA
6601 CGATACGGGA GGGCTTACCA TCTGGCCCCA GTGCTGCAAT GATACCGCGA GACCCACGCT
6661 CACCGGCTCC AGATTTATCA GCAATAAACC AGCCAGCCGG AAGGGCCGAG CGCAGAAGTG
6721 GTCCTGCAAC TTTATCCGCC TCCATCCAGT CTATTAATTG TTGCCGGGAA GCTAGAGTAA
6781 GTAGTTCGCC AGTTAATAGT TTGCGCAACG TTGTTGCCAT TGCTACAGGC ATCGTGGTGT
6841 CACGCTCGTC GTTTGGTATG GCTTCATTCA GCTCCGGTTC CCAACGATCA AGGCGAGTTA
6901 CATGATCCCC CATGTTGTGC AAAAAAGCGG TTAGCTCCTT CGGTCCTCCG ATCGTTGTCA
6961 GAAGTAAGTT GGCCGCAGTG TTATCACTCA TGGTTATGGC AGCACTGCAT AATTCTCTTA
7021 CTGTCATGCC ATCCGTAAGA TGCTTTTCTG TGACTGGTGA GTACTCAACC AAGTCATTCT
7081 GAGAATAGTG TATGCGGCGA CCGAGTTGCT CTTGCCCGGC GTCAATACGG GATAATACCG
7141 CGCCACATAG CAGAACTTTA AAAGTGCTCA TCATTGGAAA ACGTTCTTCG GGGCGAAAAC
7201 TCTCAAGGAT CTTACCGCTG TTGAGATCCA GTTCGATGTA ACCCACTCGT GCACCCAACT
7261 GATCTTCAGC ATCTTTTACT TTCACCAGCG TTTCTGGGTG AGCAAAAACA GGAAGGCAAA
7321 ATGCCGCAAA AAAGGGAATA AGGGCGACAC GGAAATGTTG AATACTCATA CTCTTCCTTT
7381 TTCAATATTA TTGAAGCATT TATCAGGGTT ATTGTCTCAT GAGCGGATAC ATATTTGAAT
7441 GTATTTAGAA AAATAAACAA ATAGGGGTTC CGCGCACATT TCCCCGAAAA GTGCCACCTG
7501 TATGCGGTGT GAAATACCGC ACAGATGCGT AAGGAGAAAA TACCGCATCA GGAAATTGTA
7561 AGCGTTAATA TTTTGTTAAA ATTCGCGTTA AATTTTTGTT AAATCAGCTC ATTTTTTAAC
7621 CAATAGGCCG AAATCGGCAA AATCCCTTAT AAATCAAAAG AATAGACCGA GATAGGGTTG
7681 AGTGTTGTTC CAGTTTGGAA CAAGAGTCCA CTATTAAAGA ACGTGGACTC CAACGTCAAA
7741 GGGCGAAAAA CCGTCTATCA AGGGCGATGG CCCACTAC
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文献和实验相关实验
1 )和转录终止子(3′AOX1 )构建成整合型表达载体。PAOX1 是一个极强的启动子，醇氧化酶的产量最高可占甲醇酵母中可溶性蛋白质的30%［2］ ，所以能使外源蛋白质在它的控制下高效产生。其次，在载体中加入酿酒酵母的分泌信号和前导肽序列(α因子)构建分泌型载体，一方面可以减轻宿主细胞的代谢负荷，另一方面可以减少宿主细胞蛋白水解酶对外源蛋白质的降解，pPIC9K、pMETαA、B、C均为此类载体。此外，使多拷贝目的基因整合入甲醇酵母染色体，形成多个表达单元，产生高产的菌株，此类载体有pAO815
4XMalE primerM13F(-47)pMAL-p2XMalE primerM13F(-47)pMET A/B/CAUG1FAUG1RpMD18-TM13R(-48)ECORIM13F(-47)(HINDⅢ)pMD19-TM13F47M13R48pMD20-TM13F47M13R48pMOSBlueT7/M13revM13forpMSCV-puropLXSNfwpMSCVrevpMT/V5-His_A(_B,_C)MtforBGH rev,V5revpNTAPT3T7pCTAPpCTAP
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pMET A
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