pVAX1哺乳表达质粒

保存条件 ：负20度

保质期 ：2年

供应商 ：钦诚生物

规格 ：5ug

基本信息

启动子:	CMV promoter
复制子:	pUC ori
终止子:	BGH poly（A）signal
质粒分类:	哺乳细胞载体;蛋白过表达质粒
质粒大小:	2999bp
原核抗性:	卡那霉素Kan
克隆菌株:	大肠杆菌DH5α
培养条件:	37℃，有氧，LB
表达宿主:	哺乳细胞293T
培养条件:	37℃，5%CO2
诱导方式:	无需诱导
5'测序引物:	CMV-F: CGCAAATGGGCGGTAGGCGTG
3'测序引物:	BGH-R: TAGAAGGCACAGTCGAGG
备注:	真核表达

质粒简介

本载体的特征是允许在大肠杆菌(Escherichia coli)中进行高拷贝复制和在哺乳动物(mammal;mammalian)细胞中高水平的瞬时表达蛋白。载体包含下列元素：
       •人巨细胞病毒（CMV）立即早期启动子，使得本载体可以在大部分的哺乳动物(mammal;mammalian)细胞中进行高水平基因表达
       •牛生长激素（BGH）多聚腺苷酸化信号能够保证高效的转录终止和mRNA的poly A结构。
       •在大肠杆菌(Escherichia coli)中是用卡那抗性进行修改载体筛选。
       •质粒pVAX1/lacZ是用来作为细胞系目的基因转染和表达的阳性对照。

质粒图谱

质粒序列

LOCUS       Exported                2999 bp ds-DNA     circular SYN 18-九月-2016
DEFINITION  synthetic circular DNA
KEYWORDS    Untitled 19
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2999)
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..2999
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        36..415
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        416..619
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        664..682
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    829..1053
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     CDS             1226..2020
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      2346..2934
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 gactcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
       61 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      121 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      181 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      241 ctatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      301 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      361 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      421 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      481 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      541 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      601 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
      661 aattaatacg actcactata gggagaccca agctggctag cgtttaaact taagcttggt
      721 accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcca gcacagtggc
      781 ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
      841 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
      901 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
      961 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     1021 gcaggcatgc tggggatgcg gtgggctcta tggcttctac tgggcggttt tatggacagc
     1081 aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt
     1141 aaactggatg gctttctcgc cgccaaggat ctgatggcgc aggggatcaa gctctgatca
     1201 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     1261 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     1321 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     1381 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     1441 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     1501 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     1561 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     1621 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     1681 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     1741 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     1801 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     1861 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     1921 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     1981 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga attattaacg cttacaattt
     2041 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacaggtggc
     2101 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2161 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatagca cgtgctaaaa
     2221 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2281 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2341 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2401 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2461 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     2521 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2581 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2641 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2701 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2761 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2821 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2881 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2941 agcaacgcgg cctttttacg gttcctgggc ttttgctggc cttttgctca catgttctt