pPICZ A
pPICZ A
pPICZ A
编号 | 载体名称 |
北京华越洋生物VECT2440 | pPICZ A |
pPICZA载体基本信息
| 载体名称: | pPICZA, pPICZ A |
|---|---|
| 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: | 3329 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: | -- |
| 稳定性: | 稳定 Stable |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
pPICZA载体质粒图谱和多克隆位点信息
pPICZA载体简介
pPICZ A，B和C是3.3 kb的毕赤酵母蛋白载体。表达的重组蛋白是融合蛋白，含有一个C-端多肽，多太重含c-myc和C-His标签。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。pPICZ系列载体包含以下元素：
•5'片段含有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）。
•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。
pPICZA载体序列
LOCUS pPICZ A 3329 bp ds-DNA circular SYN DEFINITION Pichia pastoris vector for methanol-inducible intracellular expression of a protein. ACCESSION . VERSION . KEYWORDS pPICZ A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct COMMENT This file is created by Vector NTI COMMENT Linearize with SacI, PmeI, or BstXI for integration at the AOX1 locus. FEATURES Location/Qualifiers source 1..3329 /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" misc_feature 932..1011 /note="MCS" /note="multiple cloning site" /note="color: #99ccff" CDS 1012..1041 /codon_start=1 /product="Myc (human c-Myc oncogene) epitope tag" /note="Myc" /note="color: #cc99b2" /translation="EQKLISEEDL" CDS 1057..1074 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /note="color: #cc99b2" /translation="HHHHHH" terminator 1153..1399 /gene="Pichia pastoris AOX1" /note="AOX1 terminator" /note="transcription terminator for AOX1" /note="color: #ffffff" promoter 1439..1825 /gene="S. cerevisiae TEF1" /note="TEF1 promoter" /note="promoter for EF-1-alpha" /note="color: #ffffff; direction: RIGHT" promoter 1833..1880 /note="EM7 promoter" /note="synthetic bacterial promoter?" /note="color: #ffffff; direction: RIGHT" CDS 1899..2273 /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 2339..2586 /gene="S. cerevisiae CYC1" /note="CYC1 terminator" /note="transcription terminator for CYC1" /note="color: #ffffff" rep_origin complement(2661..3249) /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 aggaattcac gtggcccagc 961 cggccgtctc ggatcggtac ctcgagccgc ggcggccgcc agcttgggcc cgaacaaaaa 1021 ctcatctcag aagaggatct gaatagcgcc gtcgaccatc atcatcatca tcattgagtt 1081 ttagccttag acatgactgt tcctcagttc aagttgggca cttacgagaa gaccggtctt 1141 gctagattct aatcaagagg atgtcagaat gccatttgcc tgagagatgc aggcttcatt 1201 tttgatactt ttttatttgt aacctatata gtataggatt ttttttgtca ttttgtttct 1261 tctcgtacga gcttgctcct gatcagccta tctcgcagct gatgaatatc ttgtggtagg 1321 ggtttgggaa aatcattcga gtttgatgtt tttcttggta tttcccactc ctcttcagag 1381 tacagaagat taagtgagac cttcgtttgt gcggatcccc cacacaccat agcttcaaaa 1441 tgtttctact ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac 1501 ttcaaaacac ccaagcacag catactaaat tttccctctt tcttcctcta gggtgtcgtt 1561 aattacccgt actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg 1621 tcgaaaaagg caataaaaat ttttatcacg tttctttttc ttgaaatttt tttttttagt 1681 ttttttctct ttcagtgacc tccattgata tttaagttaa taaacggtct tcaatttctc 1741 aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg ttcattagaa 1801 agaaagcata gcaatctaat ctaaggggcg gtgttgacaa ttaatcatcg gcatagtata 1861 tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg 1921 ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg 1981 ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc 2041 tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg 2101 tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg 2161 acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc 2221 tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtcc 2281 gacggcggcc cacgggtccc aggcctcgga gatccgtccc ccttttcctt tgtcgatatc 2341 atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa 2401 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta 2461 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac 2521 gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta 2581 atttgcaagc tggagaccaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 2641 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 2701 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 2761 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 2821 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt 2881 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 2941 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 3001 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 3061 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 3121 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 3181 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 3241 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 3301 tcacgttaag ggattttggt catgagatc //
pPICZ A其他酵母表达载体：
