pEGFP-LC3b哺乳自噬质粒

pEGFP-LC3b哺乳自噬质粒

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  • 钦诚生物
  • 上海
  • QC0199
  • 2025年07月16日
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    • 详细信息
    • 技术资料
    • 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,f1 ori,SV40 ori
    终止子: SV40 poly(A) signal
    质粒大小: 5113bp
    质粒标签: N-EGFP
    原核抗性: 卡那霉素Kan
    筛选标记: 新霉素Neo/G418/EGFP
    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
    诱导方式: 无须诱导,瞬时表达
    5'测序引物: CMV-FCGCAAATGGGCGGTAGGCGTG
    3'测序引物: Sv40-polyA-RGAAATTTGTGATGCTATTGC

    质粒简介
                pEGFP-LC3b质粒是一个哺乳细胞过表达质粒,CMV启动子驱动EGFP绿色荧光蛋白基因和LC3b人源基因融合表达,可用于细胞自噬研究。

    质粒图谱
    pEGFP-LC3b哺乳自噬质粒
    质粒序列

    LOCUS       Exported                5113 bp ds-DNA     circular SYN 29-DEC-2015
    DEFINITION  synthetic circular DNA
    KEYWORDS    pEGFP-LC3b-h
    SOURCE      synthetic DNA construct
      ORGANISM  synthetic DNA construct
    REFERENCE   1  (bases 1 to 5113)
      TITLE     Direct Submission
    FEATURES             Location/Qualifiers
         source          1..5113
                         /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"
         CDS             613..1329
                         /codon_start=1
                         /product="enhanced GFP"
                         /note="EGFP"
                         /note="mammalian codon-optimized"
                         /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                         KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                         GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                         VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                         EFVTAAGITLGMDELYK"
         misc_feature    1363..1740
                         /note="LC3B-h"
         polyA_signal    1901..2022
                         /note="SV40 poly(A) signal"
                         /note="SV40 polyadenylation signal"
         rep_origin      complement(2029..2484)
                         /direction=LEFT
                         /note="f1 ori"
                         /note="f1 bacteriophage origin of replication; arrow
                         indicates direction of (+) strand synthesis"
         promoter        2511..2615
                         /gene="bla"
                         /note="AmpR promoter"
         promoter        2617..2974
                         /note="SV40 promoter"
                         /note="SV40 enhancer and early promoter"
         rep_origin      2825..2960
                         /note="SV40 ori"
                         /note="SV40 origin of replication"
         CDS             3009..3803
                         /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    4035..4082
                         /note="HSV TK poly(A) signal"
                         /note="herpes simplex virus thymidine kinase
                         polyadenylation signal (Cole and Stacy, 1985)"
         rep_origin      4411..4999
                         /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
          661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
          721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
          781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
          841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
          901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
          961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
         1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
         1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
         1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
         1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
         1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
         1321 ctgtacaagt actcagatct cgagctcaag cttcgaattc ccatgccgtc ggagaagacc
         1381 ttcaagcagc gccgcacctt cgaacaaaga gtagaagatg tccgacttat tcgagagcag
         1441 catccaacca aaatcccggt gataatagaa cgatacaagg gtgagaagca gcttcctgtt
         1501 ctggataaaa caaagttcct tgtacctgac catgtcaaca tgagtgagct catcaagata
         1561 attagaaggc gcttacagct caatgctaat caggccttct tcctgttggt gaacggacac
         1621 agcatggtca gcgtctccac accaatctca gaggtgtatg agagtgagaa agatgaagat
         1681 ggattcctgt acatggtcta tgcctcccag gagacgttcg ggatgaaatt gtcagtgtaa
         1741 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat
         1801 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
         1861 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
         1921 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
         1981 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
         2041 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
         2101 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
         2161 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
         2221 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
         2281 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
         2341 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
         2401 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
         2461 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
         2521 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
         2581 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
         2641 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
         2701 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
         2761 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
         2821 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
         2881 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
         2941 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
         3001 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
         3061 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
         3121 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
         3181 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
         3241 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
         3301 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
         3361 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
         3421 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
         3481 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
         3541 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
         3601 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
         3661 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
         3721 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
         3781 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
         3841 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
         3901 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
         3961 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
         4021 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
         4081 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
         4141 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
         4201 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
         4261 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
         4321 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
         4381 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
         4441 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
         4501 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
         4561 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
         4621 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
         4681 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
         4741 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
         4801 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
         4861 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
         4921 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
         4981 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
         5041 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
         5101 taccgccatg cat
    //

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