pEGFP-LC3b哺乳细胞自噬绿色荧光质粒

货号 名称 规格 优惠价
P0199 pEGFP-LC3b哺乳细胞自噬绿色荧光质粒

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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-F:CGCAAATGGGCGGTAGGCGTG
3'测序引物: Sv40-polyA-R:GAAATTTGTGATGCTATTGC
            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。本质粒的参考文献共有三篇,分别是:
           1.
Crosstalk of ROS/RNS and autophagy in silibinin-induced apoptosis of MCF-7 human breast cancer cells in vitro.
          2.Type I collagen promotes primary cilia growth through down-regulating HDAC6-mediated autophagy in confluent mouse embryo fibroblast 3T3-L1 cells.
           3.Silibinin negatively contributes to primary cilia length via autophagy regulated by histone deacetylase 6 in confluent mouse embryo fibroblast 3T3-L1 cells.
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
            http://www.miaolingbio.com
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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