| 出品公司: | ZYbscience |
|---|---|
| 载体名称: | pEF1/V5-His C |
| 质粒类型: | 哺乳细胞表达载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 启动子: | EF-1α |
| 克隆方法: | 多克隆位点,限制性内切酶 |
| 载体大小: | 6184 bp |
| 5' 测序引物及序列: | T7 Forward:TAATACGACTCACTATAGGG |
| 3' 测序引物及序列: | -- |
| 载体标签: | His Tag (6x), V5 Epitope Tag |
| 载体抗性: | 氨苄青霉素(Ampicillin) |
| 筛选标记: | Geneticin |
| 备注: | 易于克隆到其它载体中去;高表达水平。 |
| 产品目录号: | ZY920-20 |
| 稳定性: | 瞬表达 |
| 组成型: | 组成型表达 |
| 病毒/非病毒: | 非病毒 |
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
-20℃低温保存
- 保质期:
三年
- 英文名:
pEF1/V5-His C
- 库存:
20
- 供应商:
泽叶生物
pEF1/V5-HisC载体基本信息
pEF1/V5-HisC载体质粒图谱和多克隆位点信息
pEF1/V5-HisC载体简介
pEF1-V5-His C
All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells,
a choice of selection marker for generating stable cell lines, and an epitope tag for easy
detection with a monoclonal antibody and rapid purification on nickel-chelating resin.
Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.
pEF1/V5-HisC载体序列
LOCUS pEF1/V5 His C 6184 bp DNA SYN
DEFINITION pEF1/V5 His C
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..6184
/organism="pEF1/V5 His C"
/mol_type="other DNA"
promoter complement(28..56)
/label="AmpR_promoter"
promoter 470..1653
/label="EF1a_promoter"
CDS complement(773..1393)
/label="ORF frame 1"
/translation="MKRRLRTERPFSFVWVTHPPALPSAASSILSSLQQGREAAIFPL
TQLVPTGPALPPRAGRYTAARGQAPEQAGQLETTPVRFSVAALAGPASPNMCAGTHGP
RRRPRPQKPKYQCADLGPHLQDYLARKKASQQVIKNFKWLETYRKQRDRREGATRFAR
GGPSAQARPQLKHEAKGLLKRKASNSPTHFQPEARDQESRTAARWK*"
misc_feature 1601..1621
/label="EF1a_fwd_primer"
/translation="MKRRLRTERPFSFVWVTHPPALPSAASSILSSLQQGREAAIFPL
TQLVPTGPALPPRAGRYTAARGQAPEQAGQLETTPVRFSVAALAGPASPNMCAGTHGP
RRRPRPQKPKYQCADLGPHLQDYLARKKASQQVIKNFKWLETYRKQRDRREGATRFAR
GGPSAQARPQLKHEAKGLLKRKASNSPTHFQPEARDQESRTAARWK*"
promoter 1670..1688
/label="T7_promoter"
/translation="MKRRLRTERPFSFVWVTHPPALPSAASSILSSLQQGREAAIFPL
TQLVPTGPALPPRAGRYTAARGQAPEQAGQLETTPVRFSVAALAGPASPNMCAGTHGP
RRRPRPQKPKYQCADLGPHLQDYLARKKASQQVIKNFKWLETYRKQRDRREGATRFAR
GGPSAQARPQLKHEAKGLLKRKASNSPTHFQPEARDQESRTAARWK*"
misc_feature complement(1894..1911)
/label="BGH_rev_primer"
/translation="MKRRLRTERPFSFVWVTHPPALPSAASSILSSLQQGREAAIFPL
TQLVPTGPALPPRAGRYTAARGQAPEQAGQLETTPVRFSVAALAGPASPNMCAGTHGP
RRRPRPQKPKYQCADLGPHLQDYLARKKASQQVIKNFKWLETYRKQRDRREGATRFAR
GGPSAQARPQLKHEAKGLLKRKASNSPTHFQPEARDQESRTAARWK*"
terminator 1897..2124
/label="bGH_PA_terminator"
/translation="MKRRLRTERPFSFVWVTHPPALPSAASSILSSLQQGREAAIFPL
TQLVPTGPALPPRAGRYTAARGQAPEQAGQLETTPVRFSVAALAGPASPNMCAGTHGP
RRRPRPQKPKYQCADLGPHLQDYLARKKASQQVIKNFKWLETYRKQRDRREGATRFAR
GGPSAQARPQLKHEAKGLLKRKASNSPTHFQPEARDQESRTAARWK*"
rep_origin 2187..2493
/label="f1_origin"
/translation="MKRRLRTERPFSFVWVTHPPALPSAASSILSSLQQGREAAIFPL
TQLVPTGPALPPRAGRYTAARGQAPEQAGQLETTPVRFSVAALAGPASPNMCAGTHGP
RRRPRPQKPKYQCADLGPHLQDYLARKKASQQVIKNFKWLETYRKQRDRREGATRFAR
GGPSAQARPQLKHEAKGLLKRKASNSPTHFQPEARDQESRTAARWK*"
misc_feature complement(2607..2627)
/label="pBABE_3_primer"
/translation="MKRRLRTERPFSFVWVTHPPALPSAASSILSSLQQGREAAIFPL
TQLVPTGPALPPRAGRYTAARGQAPEQAGQLETTPVRFSVAALAGPASPNMCAGTHGP
RRRPRPQKPKYQCADLGPHLQDYLARKKASQQVIKNFKWLETYRKQRDRREGATRFAR
GGPSAQARPQLKHEAKGLLKRKASNSPTHFQPEARDQESRTAARWK*"
misc_feature complement(2613..2829)
/label="SV40_enhancer"
/translation="MKRRLRTERPFSFVWVTHPPALPSAASSILSSLQQGREAAIFPL
TQLVPTGPALPPRAGRYTAARGQAPEQAGQLETTPVRFSVAALAGPASPNMCAGTHGP
RRRPRPQKPKYQCADLGPHLQDYLARKKASQQVIKNFKWLETYRKQRDRREGATRFAR
GGPSAQARPQLKHEAKGLLKRKASNSPTHFQPEARDQESRTAARWK*"
promoter 2625..2894
/label="SV40_promoter"
/translation="MKRRLRTERPFSFVWVTHPPALPSAASSILSSLQQGREAAIFPL
TQLVPTGPALPPRAGRYTAARGQAPEQAGQLETTPVRFSVAALAGPASPNMCAGTHGP
RRRPRPQKPKYQCADLGPHLQDYLARKKASQQVIKNFKWLETYRKQRDRREGATRFAR
GGPSAQARPQLKHEAKGLLKRKASNSPTHFQPEARDQESRTAARWK*"
rep_origin 2793..2870
/label="SV40_origin"
/translation="MKRRLRTERPFSFVWVTHPPALPSAASSILSSLQQGREAAIFPL
TQLVPTGPALPPRAGRYTAARGQAPEQAGQLETTPVRFSVAALAGPASPNMCAGTHGP
RRRPRPQKPKYQCADLGPHLQDYLARKKASQQVIKNFKWLETYRKQRDRREGATRFAR
GGPSAQARPQLKHEAKGLLKRKASNSPTHFQPEARDQESRTAARWK*"
misc_feature 2855..2874
/label="SV40pro_F_primer"
/translation="MKRRLRTERPFSFVWVTHPPALPSAASSILSSLQQGREAAIFPL
TQLVPTGPALPPRAGRYTAARGQAPEQAGQLETTPVRFSVAALAGPASPNMCAGTHGP
RRRPRPQKPKYQCADLGPHLQDYLARKKASQQVIKNFKWLETYRKQRDRREGATRFAR
GGPSAQARPQLKHEAKGLLKRKASNSPTHFQPEARDQESRTAARWK*"
CDS 3009..3803
/label="ORF frame 3"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
gene 3012..3800
/label="NeoR/KanR"
/gene="NeoR/KanR"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
terminator 3982..4101
/label="SV40_PA_terminator"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
misc_feature 4070..4089
/label="EBV_rev_primer"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
promoter complement(4145..4163)
/label="M13_reverse_primer"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
misc_feature complement(4162..4184)
/label="M13_pUC_rev_primer"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
promoter complement(4198..4227)
/label="lac_promoter"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
rep_origin complement(4536..5155)
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文献和实验广义的细胞色素 c是含有血红素 c的细胞色素之总称。通常是指主要存在于高等动植物、酵母、霉菌等线粒体中的细胞色素 c,起着传递电子的作用。为分子量约 1.3万的碱性蛋白质,还原型的吸收带为 550、 520、 415毫微米,氧化型为 407毫微米。α 因此很早便被研究,并获得结晶。不仅一级结构而且立体结构也已清楚(图 1)。其结构基本上是生物界共有的。由细胞色素 c氧化酶〔细胞色素( a+ a3 )复合体〕氧化。在呼吸链中从细胞色素 c开始接受电子,各种还原酶都已清楚
【参考值】 凝血酶原:83%~127% 因子V:50%~150% 因子VII:65%~135% 因子X:45%~155% 【临床意义】 将病人血浆分别与测定凝血酶原基质血浆(人工制备凝血酶原血浆),测定因子V用的基质血浆(储存血浆或先天性因子V缺乏症患者血浆,凝血酶原时间测定应>1分钟),测定因子VII用的基质血浆(先天性因子VII缺乏症患者的血浆。基质血浆也可采用先天性因子VII缺乏症患者血浆与正常人硫酸钡吸附血浆等量混合),测定因子X
1.常用载体分类:2,常用载体元件介绍:(1) 启动子:启动子是位于结构基因5'端上游的DNA序列,能活化RNA聚合酶,使之与模板DNA准确的结合并具有转录起始的特异性。启动子主要分为广谱启动子和特异性启动子。广谱启动子为在各种组织、细胞上广泛表达的启动子;特异性启动子为在特异性组织或者细胞类型上表达的启动子。常用广谱启动子有CAG,CMV,EF1a,PGK等,还包含启动RNA的III型启动子U6和H1启动子;常用特异性启动子如成熟神经元特异性启动子hSyn,投射神经元特异性启动
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