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pACGFP1-N1

pACGFP1-N1
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  • 产品名称:pACGFP1-N1
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简单介绍
pACGFP1-N1质粒是 AcGFP的衍生物,来自Aequorea coerulescens。 AcGFP1已经优化了更亮的荧光。pACGFP1-N1(激发*大值= 475nm;发射*大值= 505nm)AcGFP1基因的编码序列含有沉默碱基变化,其对应于人类密码子使用偏好。
产品描述

pACGFP1-N1

  • 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
XY0138 pACGFP1-N1

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

启动子: CMV promoter
复制子: pUC ori,f1 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: 4726bp
质粒标签: C-GFP
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: CMV-F(CGCAAATGGGCGGTAGGCGTG)
3'测序引物: Sv40-polyA-R (GAAATTTGTGATGCTATTGC)

质粒简介

        pAcGFP1-N1质粒是 AcGFP的衍生物,来自Aequorea coerulescens。 AcGFP1已经优化了更亮的荧光。(激发*大值= 475nm;发射*大值= 505nm)AcGFP1基因的编码序列含有沉默碱基变化,其对应于人类密码子使用偏好。
        pAcGFP1-N1 encodes a green fluorescent protein (GFP) from Aequorea coerulescens (excitation maximum = 475 nm; emission maximum = 505 nm). The coding sequence of the AcGFP1 gene
contains silent base changes, which correspond to human codon-usage preferences (1). The MCS in pAcGFP1-N1 is between the immediate early promoter of CMV (PCMV IE) and the AcGFP1 coding
sequences. Genes cloned into the MCS will be expressed as fusions to the N-terminus of AcGFP1 if they are in the same reading frame as AcGFP1 and there are no intervening stop codons. SV40
polyadenylation signals downstream of the AcGFP1 gene direct proper processing of the 3' end of theAcGFP1 mRNA. The vector backbone also contains an SV40 origin for replication in mammalian
cells expressing the SV40 T antigen.Aneomycin-resistance cassette (Neor), consisting of the SV40 earlypromoter,theneomycin/kanamycinresistancegeneofTn5,andpolyadenylationsignals fromthe
Herpes simplex virus thymidine kinase (HSV TK) gene, allows stably transfected eukaryotic cells to be selected using G418.Abacterial promoter upstream of the gene expresses kanamycin resistance
in E. coli. The pAcGFP1-N1 backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.
        Fusions to the N terminus of AcGFP1 retain the fluorescent properties of the native protein allowing the localization of the fusion protein in vivo . The target gene should be cloned into pAcGFP1-N1
so that it is in frame with the AcGFP1 coding sequences, with no intervening in-frame stop codons. The inserted gene should include the initiating ATG codon. The recombinant AcGFP1 vector can
be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418 (2). pAcGFP1-N1 can also be used simply to express
AcGFP1 in a cell line of interest (e.g., as a transfection marker).
• Human cytomegalovirus (CMV) immediate early promoter: 1–589
        Enhancer region:59–465; TATA box: 554–560
        Transcription start point: 583
        C→G mutation to remove Sac I site: 569
        • Multiple Cloning Site (MCS): 591–671
        • Aequorea coerulescens Green Fluorescent Protein (AcGFP): 673–1389
        Start codon (ATG): 673–675; Stop codon: 1390–1392
        Insertion of Val at position 2: 676–678
        Las amino acid: 1387–1389
• SV40 early mRNA polyadenylation signal
        Polyadenylation signals: 1545–1550 & 1574–1579; mRNA 3' ends: 1583 & 1595
• f1 single-strand DNA origin: 1642–2097 (Packages the noncoding strand of AcGFP)
• Bacterial promoter for expression of Kanr gene:
        –35 region: 2159–2164; –10 region: 2159–2164
        Transcription start point: 2154
• SV40 origin of replication: 2438–2573
• SV40 early promoter
        Enhancer (72-bp tandem repeats): 2271–2342 & 2343–2414
        21-bp repeats: 2418–2438, 2439–2459 & 2467–2481
        Early promoter element: 2494–2500
        Major transcription start points: 2490, 2528, 2534 & 2539
• Kanamycin/neomycin resistance gene:
        Neomycin phosphotransferase coding sequences: start codon (ATG): 2622–2624; stop codon: 3414–3416
        GA mutation to remove Pst I site: 2804
        C-A (Arg to Ser) mutation to remove BssHII site: 3150
• Herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signal
        Polyadenylation signals: 3652–3657 & 3665–3670
• pUC plasmid replication origin: 4001–4644

质粒图谱



质粒序列

LOCUS       Exported                4726 bp ds-DNA    circular SYN 18-10-2015
KEYWORDS    Untitled 2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4726)
  AUTHORS   admin
  TITLE     Direct Submission
  JOURNAL   Exported 2015-10-18  from MiaoLingPlasmid
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..4726
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    591..671
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             673..1392
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /note="AcGFP1"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    1514..1635
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1642..2097)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2124..2228
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        2230..2587
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      2438..2573
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             2622..3416
                     /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"
     polyA_signal    3648..3695
                     /note="HSV TK poly(A) signal"
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     rep_origin      4024..4612
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      661 gatccaccgg tcatggtgag caagggcgcc gagctgttca ccggcatcgt gcccatcctg
      721 atcgagctga atggcgatgt gaatggccac aagttcagcg tgagcggcga gggcgagggc
      781 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcctgtg
      841 ccctggccca ccctggtgac caccctgagc tacggcgtgc agtgcttctc acgctacccc
      901 gatcacatga agcagcacga cttcttcaag agcgccatgc ctgagggcta catccaggag
      961 cgcaccatct tcttcgagga tgacggcaac tacaagtcgc gcgccgaggt gaagttcgag
     1021 ggcgataccc tggtgaatcg catcgagctg accggcaccg atttcaagga ggatggcaac
     1081 atcctgggca ataagatgga gtacaactac aacgcccaca atgtgtacat catgaccgac
     1141 aaggccaaga atggcatcaa ggtgaacttc aagatccgcc acaacatcga ggatggcagc
     1201 gtgcagctgg ccgaccacta ccagcagaat acccccatcg gcgatggccc tgtgctgctg
     1261 cccgataacc actacctgtc cacccagagc gccctgtcca aggaccccaa cgagaagcgc
     1321 gatcacatga tctacttcgg cttcgtgacc gccgccgcca tcacccacgg catggatgag
     1381 ctgtacaagt gagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     1441 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     1501 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     1561 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     1621 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1681 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1741 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1801 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1861 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1921 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1981 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     2041 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     2101 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2161 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2221 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     2281 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2341 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2401 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2461 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     2521 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     2581 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     2641 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     2701 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     2761 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     2821 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     2881 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     2941 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3001 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3061 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3121 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     3181 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3241 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3301 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3361 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     3421 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3481 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3541 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     3601 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     3661 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     3721 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     3781 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     3841 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     3901 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3961 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4021 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4081 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4141 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     4201 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4261 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4321 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4381 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4441 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4501 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4561 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4621 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4681 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
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