| 出品公司: | Invitrogen |
|---|---|
| 载体名称: | pGAPZalpha B, pGAPZα B, pGAPZalphaB, pGAPZαB |
| 质粒类型: | 毕赤酵母蛋白表达载体 |
| 表达水平: | 高拷贝 |
| 启动子: | GAP |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 3151 bp |
| 5' 测序引物及序列: | alpha-Factor-F: 5´-TACTATTGCCAGCATTGCTGC-3´; pGAP-F:5´-GTCCCTATTTCAATCAATTGAA-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/ Zeocin |
| 备注: |
插入基因是必需包含起始密码子ATG。
|
| 产品目录号: | V200-20 |
| 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
pGAPZalpha B载体基本信息
pGAPZalpha B载体图谱质粒图谱和多克隆位点信息
pGAPZalpha B载体简介
甘油醛-3-磷酸脱氢酶（GAPDH）的酶在许多生物细胞中包括毕赤酵母，稳定高水平表达。 最近，GAPDH蛋白编码基因启动子（GAP）被深入研究，并在毕赤酵母中能够高水表达重组蛋白，这取决于使用的碳源（Waterham等 人，，1997）。GAP启动子（PGAP）的重组蛋白表达水平略高于AOX1启动子。
pGAPZ A，B，和C载体（2.9 KB）和pGAPZα，B，和C（3.1 KB）载体使用GAP启动子在毕赤酵母中稳定表达重组蛋白。重组蛋白表达为为带有C-末端myc抗原表位和C端His标签的融合蛋白。此外，pGAPZα 表达的融合蛋白在N-端带有酿酒酵母（Saccharomyces cerevisiae）的α-因子分泌信号肽。这两个系列的载体都提供三个不同读码框的载体，以方便克隆的带有C-端标签和/或N-末端分泌信号肽的载体 中。
这些载体是基于显性选择标记，Zeocin（博莱霉素）的筛选标记可以同时使用于毕赤酵母和大肠杆菌。
pGAPZalpha B载体序列
LOCUS pGAPZα B 3151 bp ds-DNA circular SYN 05-二月-2013
DEFINITION Pichia pastoris vector for constitutive expression of a secreted
protein.
ACCESSION .
VERSION .
KEYWORDS pGAPZ(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 within the GAP promoter for integration.
FEATURES Location/Qualifiers
source 1..3151
/organism="synthetic DNA construct"
/lab_host="Pichia pastoris"
/mol_type="other DNA"
promoter 7..483
/note="GAP promoter"
/note="promoter for Pichia pastoris
glyceraldehyde-3-phosphate dehydrogenase"
/note="color: #ffffff; direction: RIGHT"
CDS 493..759
/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:?493?..?549?/?#993366?/?presequence
???2:?550?..?747?/?#993366?/?pro?region
???3:?748?..?759?/?#993366
Cleavage sites after bases 549, 747"
/translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
misc_feature 758..832
/note="MCS"
/note="multiple cloning site"
/note="color: #99ccff"
CDS 831..860
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/note="color: #cc99b2"
/translation="EQKLISEEDL"
CDS 876..893
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/note="color: #cc99b2"
/translation="HHHHHH"
terminator 972..1218
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
/note="color: #ffffff"
promoter 1233..1644
/gene="S. cerevisiae TEF1"
/note="TEF1 promoter"
/note="promoter for?EF-1-alpha"
/note="color: #ffffff; direction: RIGHT"
promoter 1651..1698
/note="EM7 promoter"
/note="synthetic bacterial promoter?"
/note="color: #ffffff; direction: RIGHT"
CDS 1717..2091
/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 2157..2404
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
/note="color: #ffffff"
rep_origin complement(2479..3067)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/note="color: #ffff00"
ORIGIN
1 agatcttttt tgtagaaatg tcttggtgtc ctcgtccaat caggtagcca tctctgaaat
61 atctggctcc gttgcaactc cgaacgacct gctggcaacg taaaattctc cggggtaaaa
121 cttaaatgtg gagtaatgga accagaaacg tctcttccct tctctctcct tccaccgccc
181 gttaccgtcc ctaggaaatt ttactctgct ggagagcttc ttctacggcc cccttgcagc
241 aatgctcttc ccagcattac gttgcgggta aaacggaggt cgtgtacccg acctagcagc
301 ccagggatgg aaaagtcccg gccgtcgctg gcaataatag cgggcggacg catgtcatga
361 gattattgga aaccaccaga atcgaatata aaaggcgaac acctttccca attttggttt
421 ctcctgaccc aaagacttta aatttaattt atttgtccct atttcaatca attgaacaac
481 tatttcgaaa cgatgagatt tccttcaatt tttactgctg ttttattcgc agcatcctcc
541 gcattagctg ctccagtcaa cactacaaca gaagatgaaa cggcacaaat tccggctgaa
601 gctgtcatcg gttactcaga tttagaaggg gatttcgatg ttgctgtttt gccattttcc
661 aacagcacaa ataacgggtt attgtttata aatactacta ttgccagcat tgctgctaaa
721 gaagaagggg tatctctcga gaaaagagag gctgaagctg caggaattca cgtggcccag
781 ccggccgtct cggatcggta cctcgagccg cggcggccgc cagctttcta gaacaaaaac
841 tcatctcaga agaggatctg aatagcgccg tcgaccatca tcatcatcat cattgagttt
901 tagccttaga catgactgtt cctcagttca agttgggcac ttacgagaag accggtcttg
961 ctagattcta atcaagagga tgtcagaatg ccatttgcct gagagatgca ggcttcattt
1021 ttgatacttt tttatttgta acctatatag tataggattt tttttgtcat tttgtttctt
1081 ctcgtacgag cttgctcctg atcagcctat ctcgcagctg atgaatatct tgtggtaggg
1141 gtttgggaaa atcattcgag tttgatgttt ttcttggtat ttcccactcc tcttcagagt
1201 acagaagatt aagtgagacc ttcgtttgtg cggatccccc acacaccata gcttcaaaat
1261 gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact
1321 tcaaaacacc caagcacagc atactaaatt ttccctcttt cttcctctag ggtgtcgtta
1381 attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt
1441 cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaattttt ttttttagtt
1501 tttttctctt tcagtgacct ccattgatat ttaagttaat aaacggtctt caatttctca
1561 agtttcagtt tcatttttct tgttctatta caactttttt tacttcttgt tcattagaaa
1621 gaaagcatag caatctaatc taagggcggt gttgacaatt aatcatcggc atagtatatc
1681 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt
1741 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
1801 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
1861 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
1921 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
1981 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
2041 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtccga
2101 cggcggccca cgggtcccag gcctcggaga tccgtccccc ttttcctttg tcgatatcat
2161 gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa
2221 ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt
2281 attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc
2341 atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat
2401 ttgcaagctg gagaccaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
2461 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
2521 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
2581 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
2641 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
2701 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
2761 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
2821 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
2881 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
2941 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
3001 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
3061 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
3121 acgttaaggg attttggtca tgcatgagat c
//
