| 出品公司: | Invitrogen |
|---|---|
| 载体名称: | pPICZalpha B, pPICZα B, pPICZalphaB, pPICZαB |
| 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: | 3597 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: | V195-20 |
| 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
pPICZα B载体 pPICZalpha B质粒基本信息
pPICZα B质粒图谱 pPICZalpha B载体图谱和pPICZα B载体序列 pPICZalpha B质粒序列多克隆位点信息
pPICZα B质粒 pPICZalpha B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。
pPICZα B质粒序列 pPICZalpha B载体序列
LOCUS pPICZα B 3597 bp ds-DNA circular SYN 05-二月-2013
DEFINITION Pichia pastoris vector for methanol-inducible expression of a
secreted protein.
ACCESSION .
VERSION .
KEYWORDS pPICZ(alpha) B
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..3597
/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 1206..1280
/note="MCS"
/note="multiple cloning site"
/note="color: #99ccff"
CDS 1279..1308
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/note="color: #cc99b2"
/translation="EQKLISEEDL"
CDS 1324..1341
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/note="color: #cc99b2"
/translation="HHHHHH"
terminator 1421..1667
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
/note="color: #ffffff"
promoter 1707..2093
/gene="S. cerevisiae TEF1"
/note="TEF1 promoter"
/note="promoter for EF-1-alpha"
/note="color: #ffffff; direction: RIGHT"
promoter 2101..2148
/note="EM7 promoter"
/note="synthetic bacterial promoter?"
/note="color: #ffffff; direction: RIGHT"
CDS 2167..2541
/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 2607..2854
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
/note="color: #ffffff"
rep_origin complement(2929..3517)
/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 aaagagaggc
1201 tgaagctgca ggaattcacg tggcccagcc ggccgtctcg gatcggtacc tcgagccgcg
1261 gcggccgcca gctttctaga acaaaaactc atctcagaag aggatctgaa tagcgccgtc
1321 gaccatcatc atcatcatca ttgagtttgt agccttagac atgactgttc ctcagttcaa
1381 gttgggcact tacgagaaga ccggtcttgc tagattctaa tcaagaggat gtcagaatgc
1441 catttgcctg agagatgcag gcttcatttt tgatactttt ttatttgtaa cctatatagt
1501 ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc
1561 tcgcagctga tgaatatctt gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt
1621 tcttggtatt tcccactcct cttcagagta cagaagatta agtgagacct tcgtttgtgc
1681 ggatccccca cacaccatag cttcaaaatg tttctactcc ttttttactc ttccagattt
1741 tctcggactc cgcgcatcgc cgtaccactt caaaacaccc aagcacagca tactaaattt
1801 tccctctttc ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa
1861 aagagaccgc ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt
1921 tctttttctt gaaatttttt tttttagttt ttttctcttt cagtgacctc cattgatatt
1981 taagttaata aacggtcttc aatttctcaa gtttcagttt catttttctt gttctattac
2041 aacttttttt acttcttgtt cattagaaag aaagcatagc aatctaatct aaggggcggt
2101 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
2161 taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga
2221 gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc
2281 gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg
2341 ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg
2401 tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc
2461 gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc
2521 gtggccgagg agcaggactg acacgtccga cggcggccca cgggtcccag gcctcggaga
2581 tccgtccccc ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg
2641 ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
2701 ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt
2761 tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga
2821 gaaggttttg ggacgctcga aggctttaat ttgcaagctg gagaccaaca tgtgagcaaa
2881 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
2941 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
3001 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
3061 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
3121 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
3181 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
3241 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
3301 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
3361 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
3421 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
3481 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
3541 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagatc
//
