| 出品公司: | Invitrogen |
|---|---|
| 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: | Zeocin 博来霉素 |
| 备注: |
插入基因是必需包含起始密码子ATG。
|
| 产品目录号: | V190-20 |
| 稳定性: | 稳定 Stable |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
买家导航
pPICZ A载体 pPICZA质粒基本信息
pPICZ A质粒图谱 pPICZA载体图谱和pPICZ A载体序列 pPICZA质粒序列多克隆位点信息
pPICZ A质粒 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。
pPICZ A质粒序列 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
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..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
//
