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pEGFP-1

pEGFP-1
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  • 产品名称：pEGFP-1
  • 产品型号：
  • 产品展商：XYbscience
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简单介绍
pEGFP-1编码野生型GFP（1-3）的红移型变体，其已经针对更明亮的荧光和哺乳动物细胞中更高的表达进行了优化。pEGFP-1（激发*大值= 488nm;发射*大值= 507nm）pEGFP-1骨架还提供了用于在大肠杆菌中繁殖的pUC起始点和用于单链DNA生���的f1起源。
产品描述

pEGFP-1

  • 产品信息
  • 基本信息
  • 质粒简介
  • 质粒图谱
  • 质粒序列

产品信息

产品货号 产品名称 产品规格 优惠价
XY0132 pEGFP-1

5μg

¥请询价

使用说明

信裕质粒平台的各批次质粒菌株发货前均经过严格的多重验证，如存在质量问题，请在收到产品的三个月内通知我司。收到质粒后请短暂离心，加入20μl ddH2O溶解质粒，取2μl转化至对应感受态中，挑取单克隆重新提取质粒后使用。如需获取其他详细信息请登录我司官网查询。

基本信息

复制子: F1 ori , pUC ori
终止子: SV40 poly(A) signal
质粒分类: 哺乳系列质粒；哺乳荧光质粒；哺乳绿色质粒
质粒大小: 4151bp
质粒标签: C-EGFP
原核抗性: 卡那霉素Kan（50μg/ml）
筛选标记: 新霉素Neo/G418
克隆菌株: DH5α等大肠杆菌
培养条件: 37℃，有氧 LB
表达宿主: 293T等哺乳细胞
培养条件: 37℃，5%CO2
诱导方式: 无须诱导，瞬时表达
5'测序引物: EGFP-N（CGTCGCCGTCCAGCTCGACCAG）
3'测序引物: EGFP-F (CCAGCAACGCGGCCTTTTTA)

质粒简介

         pEGFP-1编码野生型GFP（1-3）的红移型变体，其已经针对更明亮的荧光和哺乳动物细胞中更高的表达进行了优化。（激发*大值= 488nm;发射*大值= 507nm）pEGFP-1骨架还提供了用于在大肠杆菌中繁殖的pUC起始点和用于单链DNA生产的f1起源。
        pEGFP-1 encodes a red-shifted variant of wild-type GFP (1–3) which has been optimized for brighter fluorescence and higher expression in mammalian cells. (Excitation maximum = 488 nm; emission maximum = 507 nm.) pEGFP-1 encodes the GFPmut1 variant (4) which contains the double-aminoacid substitution of Phe-64 to Leu and Ser-65 to Thr. The coding sequence of the EGFP gene contains more than 190 silent base changes which correspond to human codon-usage preferences (5). Sequences flanking EGFP have been converted to a Kozak consensus translation initiation site (6) to further increase the translation efficiency in eukaryotic cells. pEGFP-1 is a promoterless EGFP vector which can be used to monitor transcription from different promoters and promoter/enhancer combinations inserted into the MCS located upstream of the EGFP coding sequence. SV40 polyadenylation signals downstream of the EGFP gene direct proper processing of the 3' end of the EGFP mRNA. The vector backbone also contains an SV40 origin for replication in mammalian cells expressing the SV40 T antigen. A neomycin-resistance cassette (Neor) allows stably transfected eukaryotic cells to be selected using G418. The Neor cassette consists of the SV40 early promoter, the neomycin/kanamycin resistance gene of Tn5, and polyadenylation signals from the Herpes simplex virus thymidine kinase (HSV TK) gene. A bacterial promoter upstream of this cassette confers kanamycin resistance in E. coli. The pEGFP-1 backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.
       EGFP can be used as an in vivo reporter of gene expression . Promoters should be cloned into the pEGFP-1 MCS upstream from the EGFP coding sequences. Without the addition of a functional promoter, this vector will not express EGFP. The recombinant EGFP vector can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418 .

质粒图谱



质粒序列

LOCUS       Exported                4151 bp ds-DNA     circular SYN 30-AUG-2016
DEFINITION  synthetic circular DNA
KEYWORDS    Untitled
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4151)
  AUTHORS   admin
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, August 30, 2016 from MiaoLingPlasmid
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..4151
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    12..89
                     /note="MCS"
                     /note="MCS;multiple cloning site"
     CDS             97..816
                     /codon_start=1
                     /product="enhanced GFP"
                     /note="enhanced GFP"
                     /note="EGFP;mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    939..1060
                     /note="SV40 poly(A) signal"
                     /note="SV40 poly(A) signal;SV40 polyadenylation signal"
     rep_origin      complement(1067..1522)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 ori;f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1549..1653
                     /gene="bla"
                     /note="bla AmpR promoter"
                     /note="AmpR promoter"
     promoter        1655..2012
                     /note="SV40 promoter"
                     /note="SV40 promoter;SV40 enhancer and early promoter"
     rep_origin      1863..1998
                     /note="SV40 ori"
                     /note="SV40 ori;SV40 origin of replication"
     CDS             2047..2841
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="aph(3')-II (or nptII)"
                     /note="NeoR/KanR;confers resistance to neomycin, kanamycin,
                     and G418 (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3073..3120
                     /note="HSV TK poly(A) signal"
                     /note="HSV TK poly(A) signal;herpesvirus thymidine kinase 
                     polyadenylation signal"
     rep_origin      3449..4037
                     /direction=RIGHT
                     /note="ori"
                     /note="ori;high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 tagttattac tagcgctacc ggactcagat ctcgagctca agcttcgaat tctgcagtcg
       61 acggtaccgc gggcccggga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg
      121 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      181 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
      241 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
      301 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
      361 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
      421 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
      481 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
      541 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
      601 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
      661 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
      721 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
      781 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa
      841 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
      901 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
      961 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1021 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     1081 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     1141 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     1201 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     1261 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     1321 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     1381 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     1441 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     1501 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     1561 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1621 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     1681 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     1741 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     1801 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     1861 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     1921 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     1981 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     2041 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2101 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     2161 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     2221 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     2281 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     2341 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     2401 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     2461 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     2521 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     2581 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     2641 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     2701 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     2761 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     2821 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     2881 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     2941 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3001 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3061 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     3121 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     3181 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     3241 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     3301 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3361 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3421 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3481 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3541 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3601 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3661 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3721 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3781 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3841 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3901 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3961 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4021 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4081 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4141 ccgccatgca t
//
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