| 出品公司: | ZYbscience |
|---|---|
| 载体名称: | pPICZalpha C, pPICZα C, pPICZalphaC, pPICZαC |
| 质粒类型: | 毕赤酵母蛋白表达载体 |
| 表达水平: | 高拷贝 |
| 启动子: | AOX1 |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 3598 bp |
| 5' 测序引物及序列: | 5´ AOX1:5´-GACTGGTTCCAATTGACAAGC-3´ |
| 3' 测序引物及序列: | 3´ AOX1:5´-GCAAATGGCATTCTGACATCC-3´ |
| 载体标签: | C-Myc, C-His |
| 载体抗性: | Zeocin 博来霉素 |
| 筛选标记: | HIS4 |
| 备注: |
插入基因是必需包含起始密码子ATG。
|
| 产品目录号: | ZY195-20 |
| 稳定性: | 稳定 Stable |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
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保存条件 ：-20℃低温保存
保质期 ：三年
英文名 ：pPICZα C
库存 ：20
供应商 ：泽叶生物
pPICZalpha C载体基本信息
pPICZalpha C载体质粒图谱和多克隆位点信息
pPICZalpha C载体简介
pPICZα A，B和C是3.6 kb的毕赤酵母蛋白分泌表达载体。表达的重组蛋白是融合蛋白，含有一个N-端多肽，编码酿酒酵母（Saccharomyces cerevisiae）α-因子分泌信号。载体能够在毕赤酵母中利用甲醇诱导的高水 平的表达目的蛋白，并且可以用在任何毕赤酵母中，包括X33，GS115菌株，SMD1168H，KM71H。
pPICZα A，B和C系列载体包含以下元素：
•5'端含有AOX1启动子的严格调控，利用甲醇诱导表达任何感兴趣的基因（Ellis等，1985; Koutz等人，1989;tschopp等人，1987A）。
•α-因子分泌信号能够分泌性表达目的蛋白。
•Zeocin抗性基因在大肠杆菌和毕赤酵母都能用于筛选（Baron等人,1992; Drocourt等人，1990）。
•C-端含Myc和His标签，可以用于检测和纯化重组蛋白。
•pPICZ A, B, C三种读码框使得可以将基因克隆入载体而不发生任何移码突变。
pPICZalpha C载体序列
LOCUS pPICZα C 3598 bp ds-DNA circular SYN 05-二月-2013DEFINITION Pichia pastoris vector for methanol-inducible expression of a secreted protein.ACCESSION .VERSION .KEYWORDS pPICZ(alpha) CSOURCE synthetic DNA construct ORGANISM synthetic DNA constructCOMMENT This file is created by Vector NTI http://www.biofeng.com/COMMENT VNTAUTHORNAME|biofeng.com|COMMENT Linearize with SacI, PmeI, or BstXI for integration at the AOX1 locus.FEATURES Location/Qualifiers source 1..3598 /organism="synthetic DNA construct" /lab_host="Pichia pastoris" /mol_type="other DNA" promoter 2..940 /gene="Pichia pastoris AOX1" /note="AOX1 promoter" /note="inducible promoter, regulated by methanol" /note="color: #ffffff; direction: RIGHT" 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." /note="This feature has 3 segments: ???1:??941?..??997?/?#993366?/?presequence ???2:??998?..?1195?/?#993366?/?pro?region ???3:?1196?..?1207?/?#993366 Cleavage sites after bases 997, 1195" /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA" misc_feature 1207..1281 /note="MCS" /note="multiple cloning site" /note="color: #99ccff" CDS 1280..1309 /codon_start=1 /product="Myc (human c-Myc oncogene) epitope tag" /note="Myc" /note="color: #cc99b2" /translation="EQKLISEEDL" CDS 1325..1342 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /note="color: #cc99b2" /translation="HHHHHH" terminator 1422..1668 /gene="Pichia pastoris AOX1" /note="AOX1 terminator" /note="transcription terminator for AOX1" /note="color: #ffffff" promoter 1708..2094 /gene="S. cerevisiae TEF1" /note="TEF1 promoter" /note="promoter for EF-1-alpha" /note="color: #ffffff; direction: RIGHT" promoter 2102..2149 /note="EM7 promoter" /note="synthetic bacterial promoter?" /note="color: #ffffff; direction: RIGHT" CDS 2168..2542 /codon_start=1 /gene="Sh?ble from?Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /note="BleoR" /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /note="color: #ccffcc" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" terminator 2608..2855 /gene="S. cerevisiae CYC1" /note="CYC1 terminator" /note="transcription terminator for CYC1" /note="color: #ffffff" rep_origin complement(2930..3518) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00"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 agagagaggc 1201 tgaagcatcg atgaattcac gtggcccagc cggccgtctc ggatcggtac ctcgagccgc 1261 ggcggccgcc agctttctag aacaaaaact catctcagaa gaggatctga atagcgccgt 1321 cgaccatcat catcatcatc attgagtttg tagccttaga catgactgtt cctcagttca 1381 agttgggcac ttacgagaag accggtcttg ctagattcta atcaagagga tgtcagaatg 1441 ccatttgcct gagagatgca ggcttcattt ttgatacttt tttatttgta acctatatag 1501 tataggattt tttttgtcat tttgtttctt ctcgtacgag cttgctcctg atcagcctat 1561 ctcgcagctg atgaatatct tgtggtaggg gtttgggaaa atcattcgag tttgatgttt 1621 ttcttggtat ttcccactcc tcttcagagt acagaagatt aagtgagacc ttcgtttgtg 1681 cggatccccc acacaccata gcttcaaaat gtttctactc cttttttact cttccagatt 1741 ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc caagcacagc atactaaatt 1801 ttccctcttt cttcctctag ggtgtcgtta attacccgta ctaaaggttt ggaaaagaaa 1861 aaagagaccg cctcgtttct ttttcttcgt cgaaaaaggc aataaaaatt tttatcacgt 1921 ttctttttct tgaaattttt ttttttagtt tttttctctt tcagtgacct ccattgatat 1981 ttaagttaat aaacggtctt caatttctca agtttcagtt tcatttttct tgttctatta 2041 caactttttt tacttcttgt tcattagaaa gaaagcatag caatctaatc taaggggcgg 2101 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa 2161 ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg 2221 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt 2281 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt 2341 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg 2401 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg 2461 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt 2521 cgtggccgag gagcaggact gacacgtccg acggcggccc acgggtccca ggcctcggag 2581 atccgtcccc cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac 2641 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg 2701 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt 2761 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg 2821 agaaggtttt gggacgctcg aaggctttaa tttgcaagct ggagaccaac atgtgagcaa 2881 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 2941 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 3001 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 3061 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 3121 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 3181 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 3241 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 3301 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 3361 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 3421 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 3481 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 3541 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagatc//
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