基本信息
別稱:pPICZaA
啟動(dòng)子:AOX1
復(fù)制子:pUC
終止子:AOX1 TT
質(zhì)粒分類:酵母系列，畢赤酵母表達(dá)載體
質(zhì)粒大小:3593bp
質(zhì)粒標(biāo)簽:C-Myc, C-His
原核抗性:Zeo
篩選標(biāo)記:Zeo
克隆菌株:DH5a
培養(yǎng)條件:37度
表達(dá)宿主:酵母細(xì)胞
誘導(dǎo)方式:甲醇誘導(dǎo)
5'測(cè)序引物:AOX5:GACTGGTTCCAATTGACAAGC
3'測(cè)序引物:AOX3:GGCAAATGGCATTCTGACAT
質(zhì)粒屬性
載體宿主：酵母菌
載體用途：蛋白表達(dá)
基因種屬：空載體
基因類型：ORF
原核抗性：Zeo
真核抗性：Zeo
熒光蛋白：
質(zhì)粒簡(jiǎn)介
pPICZαA，B和C是3.6 kb的畢赤酵母蛋白分泌表達(dá)載體。表達(dá)的重組蛋白是融合蛋白，含有一個(gè)N-端多肽，編碼釀酒酵母（Saccharomyces cerevisiae）α-因子分泌信號(hào)。載體能夠在畢赤酵母中利用甲醇誘導(dǎo)的高水 平的表達(dá)目的蛋白，并且可以用在任何畢赤酵母中，包括X33，GS115菌株，SMD1168H，KM71H。pPICZα A，B和C系列載體包含以下內(nèi)容：
①5'端含有AOX1啟動(dòng)子的嚴(yán)格調(diào)控，利用甲醇誘導(dǎo)表達(dá)任何感興趣的基因（Ellis等，1985; Koutz等人，1989;tschopp等人，1987A）。
②α-因子分泌信號(hào)能夠分泌性表達(dá)目的蛋白。
③Zeocin抗性基因在大腸桿菌和畢赤酵母都能用于篩選（Baron等人,1992; Drocourt等人，1990）。
④C-端含Myc和His標(biāo)簽，可以用于檢測(cè)和純化重組蛋白。
⑤pPICZ A, B, C三種讀碼框使得可以將基因克隆入載體而不發(fā)生任何移碼突變。
質(zhì)粒序列
LOCUS Exported 3593 bp ds-DNA circular SYN 10-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3593)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..3593
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 2..940
/gene="Pichia pastoris AOX1"
/note="AOX1 promoter"
/note="inducible promoter, regulated by methanol"
CDS 941..1207
/codon_start=1
/gene="MF-alpha-1"
/product="N-terminal secretion signal from S. cerevisiae
alpha-factor"
/note="alpha-factor secretion signal"
/note="Cleavage by the Kex2 protease occurs after the
dibasic KR sequence. The EA dipeptides are then removed by
dipeptidyl aminopeptidase A."
/translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
CDS 1275..1304
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/translation="EQKLISEEDL"
CDS 1320..1337
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 1417..1663
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
promoter 1678..2089
/gene="S. cerevisiae TEF1"
/note="TEF1 promoter"
/note="promoter for EF-1-alpha"
promoter 2097..2144
/note="EM7 promoter"
/note="synthetic bacterial promoter "
CDS 2163..2537
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/note="BleoR"
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
terminator 2603..2850
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
rep_origin complement(2925..3513)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
901 caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt
961 tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga
1021 agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga
1081 tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa
1141 tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaagagaggc
1201 tgaagctgaa ttcacgtggc ccagccggcc gtctcggatc ggtacctcga gccgcggcgg
1261 ccgccagctt tctagaacaa aaactcatct cagaagagga tctgaatagc gccgtcgacc
1321 atcatcatca tcatcattga gtttgtagcc ttagacatga ctgttcctca gttcaagttg
1381 ggcacttacg agaagaccgg tcttgctaga ttctaatcaa gaggatgtca gaatgccatt
1441 tgcctgagag atgcaggctt catttttgat acttttttat ttgtaaccta tatagtatag
1501 gatttttttt gtcattttgt ttcttctcgt acgagcttgc tcctgatcag cctatctcgc
1561 agctgatgaa tatcttgtgg taggggtttg ggaaaatcat tcgagtttga tgtttttctt
1621 ggtatttccc actcctcttc agagtacaga agattaagtg agaccttcgt ttgtgcggat
1681 cccccacaca ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc
1741 ggactccgcg catcgccgta ccacttcaaa acacccaagc acagcatact aaattttccc
1801 tctttcttcc tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga
1861 gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt
1921 tttcttgaaa tttttttttt tagttttttt ctctttcagt gacctccatt gatatttaag
1981 ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact
2041 ttttttactt cttgttcatt agaaagaaag catagcaatc taatctaagg ggcggtgttg
2101 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
2161 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
2221 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
2281 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
2341 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
2401 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
2461 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
2521 ccgaggagca ggactgacac gtccgacggc ggcccacggg tcccaggcct cggagatccg
2581 tccccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct
2641 ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
2701 atttattttt ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt
2761 ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag
2821 gttttgggac gctcgaaggc tttaatttgc aagctggaga ccaacatgtg agcaaaaggc
2881 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
2941 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
3001 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
3061 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
3121 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
3181 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
3241 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
3301 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
3361 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
3421 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
3481 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
3541 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag atc
//
