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pCDNA3.1(+)哺乳表达质粒 货号:P2004
质粒分类: 哺乳系列质粒;哺乳表达质粒;pCDNA系列质粒
质粒大小: 5428bp
原核抗性: 氨苄青霉素Ampicillin(100μg/ml)
筛选标记: 新霉素Neomycin/G418
产品信息
别名 pCDNA3.1+
启动子 CMV
复制子 pUC
终止子 BGH poly(A) signal
质粒分类 哺乳系列质粒;哺乳表达质粒;pCDNA系列质粒
质粒大小 5428bp
原核抗性 Amp
真核抗性 G418
克隆菌株 DH5α
培养条件 37℃
表达宿主 293T等哺乳细胞
培养条件 37℃,5%CO2
诱导方式 无需诱导
5'测序引物 pCDNA3.1-F(CTAGAGAACCCACTGCTTAC)
3'测序引物 pCDNA3.1-R(TAGAAGGCACAGTCGAGG)
质粒宿主 哺乳细胞
质粒用途 蛋白表达
片段类型 基因CDS
片段物种 空载体
原核抗性 氨苄青霉素

质粒简述
        pCDNA3.1(+)是来自pCDNA3的5.4kb载体,设计用于哺乳动物宿主中的高水平稳定和瞬时表达。
大多数哺乳动物细胞可以进行高水平的稳定和非复制性瞬时表达。人类巨细胞病毒立即早期(CMV)
启动子,用于在广泛哺乳动物细胞中的高水平表达。前向(+)和反向(-)方向上的多个克隆位点,
以促进克隆用于选择稳定细胞系的新霉素抗性基因。在潜伏感染SV40或表达SV40大T抗原(例如COS-1,
COS-7)的细胞系中的异常复制。

最常用的哺乳动物表达载体之一,CMV promoter强启动子调控外源基因表达。

载体拷贝数很高,表达量也很高。

无荧光标记,也没有携带tag。

Amp+原核筛选抗性,Neo+真核筛选抗性,可以利用G418筛选稳定细胞株


       pcDNA3.1(+) and pcDNA3.1(-) are 5.4 kb vectors derived from pcDNA3 and designed for high-level stable and transient expression in mammalian hosts. High-level stable and non-replicative transient expression can be carried out in most mammalian cells. The vectors contain the following elements:
       • Human cytomegalovirus immediate-early (CMV) promoter for high-level expression in a wide range of mammalian cells
       • Multiple cloning sites in the forward (+) and reverse (-) orientations to facilitate cloning

       • Neomycin resistance gene for selection of stable cell lines
       • Episomal replication in cells lines that are latently infected with SV40 or that express the SV40 large T antigen (e.g. COS-1, COS-7). The control plasmid, pcDNA3.1/CAT, is included for use as a positive control for transfection and expression in the cell line of choice.
       This pcDNA3.1(+)vector is designed for high-level, constitutive expression in a variety of mammalian cell lines. It contains a Geneticin® selectable marker and a forward-orientation multiple cloning site.


图谱


ID   pcDNA3.1(+) standard; circular DNA; SYN; 5428 BP.
DT   28-AUG-2003
CC   This file is created by Vector NTI
CC   http://www.informaxinc.com/
CC   ORIGDB|GenBank
CC   VNTDATE|38836800|
CC   VNTDBDATE|595081614|
CC   LSOWNER|
CC   VNTAUTHORNAME|Demo User|
FH   Key             Location/Qualifiers
FH
FT   source          1..5428
FT                   /invitrogen="124"
FT                   /vntifkey="98"
FT   CDS             complement(4432..5292)
FT                   /invitrogen="1020000"
FT                   /vntifkey="4"
FT                   /label=Amp(R)
FT   polyA_site      1028..1252
FT                   /invitrogen="1870000"
FT                   /vntifkey="26"
FT                   /label=BGHpA
FT   primer          complement(1022..1039)
FT                   /invitrogen="1910000"
FT                   /vntifkey="27"
FT                   /label=BGHreverseprimer
FT   promoter        complement(5293..5391)
FT                   /invitrogen="2330000"
FT                   /vntifkey="30"
FT                   /label=blapromoter
FT   primer          769..789
FT                   /invitrogen="1930000"
FT                   /vntifkey="27"
FT                   /label=CMVforwardprimer
FT   promoter        232..819
FT                   /invitrogen="2180000"
FT                   /vntifkey="29"
FT                   /label=CMVpromoter
FT   rep_origin      1298..1726
FT                   /invitrogen="2500000"
FT                   /vntifkey="33"
FT                   /label=f1origin
FT   misc_feature    895..1010
FT                   /invitrogen="1630000"
FT                   /vntifkey="21"
FT                   /label=MCS
FT   CDS             2136..2930
FT                   /invitrogen="1260000"
FT                   /vntifkey="4"
FT                   /label=Neo(R)
FT   rep_origin      complement(3617..4287)
FT                   /invitrogen="2540000"
FT                   /vntifkey="33"
FT                   /label=pUCorigin
FT   promoter        1731..2074
FT                   /invitrogen="2290000"
FT                   /vntifkey="29"
FT                   /label=SV40earlypromoter
FT   polyA_signal    3104..3234
FT                   /invitrogen="1840000"
FT                   /vntifkey="25"
FT                   /label=SV40pA
SQ   Sequence 5428 BP; 1251 A; 1411 C; 1386 G; 1380 t;
     gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg        60
     ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg       120
     cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc       180
     ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt       240
     gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata       300
     tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc       360
     cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc       420
     attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt       480
     atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt       540
     atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca       600
     tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg       660
     actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc       720
     aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg       780
     gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca       840
     ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc       900
     gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc       960
     agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca      1020
     gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc      1080
     ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg      1140
     cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg      1200
     gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag      1260
     gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta      1320
     agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg      1380
     cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa      1440
     gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc      1500
     aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt      1560
     cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca      1620
     acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc      1680
     tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg      1740
     tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca      1800
     tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa      1860
     gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca      1920
     tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt      1980
     ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag      2040
     gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg      2100
     gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg      2160
     caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa      2220
     tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg      2280
     tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt      2340
     ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa      2400
     gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc      2460
     ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg      2520
     ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg      2580
     aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg      2640
     aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg      2700
     gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact      2760
     gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg      2820
     ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc      2880
     ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct      2940
     ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac      3000
     cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat      3060
     cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc      3120
     ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc      3180
     actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc      3240
     gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg      3300
     ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg      3360
     tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc      3420
     gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt      3480
     gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct      3540
     gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga      3600
     taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc      3660
     cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg      3720
     ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg      3780
     aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt      3840
     tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt      3900
     gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg      3960
     cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact      4020
     ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt      4080
     cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct      4140
     gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac      4200
     cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca      4260
     agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta      4320
     agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa      4380
     atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg      4440
     cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg      4500
     actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc      4560
     aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc      4620
     cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa      4680
     ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc      4740
     cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg      4800
     ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc      4860
     cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat      4920
     ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg      4980
     tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc      5040
     ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg      5100
     aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat      5160
     gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg      5220
     gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg      5280
     ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct      5340
     catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac      5400
     atttccccga aaagtgccac ctgacgtc                                         5428


		

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