pEGFP-LC3b-m哺乳自噬质粒

基本信息

启动子:	CMV
复制子:	pUC ori,f1 ori
终止子:	SV40 poly(A) signal
质粒分类:	哺乳细胞，自噬表达载体
质粒大小:	5073bp
原核抗性:	Kan
筛选标记:	Neo
克隆菌株:	DH5α
培养条件:	37℃，有氧 LB
表达宿主:	哺乳细胞
诱导方式:	无须诱导，瞬时表达
5'测序引物:	pEGFP-C-5:CATGGTCCTGCTGGAGTTCGTG
3'测序引物:	根据序列设计引物

质粒简介

      PEGFP-LC3b é um estudo para células de mamíferos autophagy FOI com expressão EM células de mamíferos, e muitas pessoas, a expressão de genes LC3b Na Fonte, Ao Mesmo tempo, Pode ser a expressão de UMA proteína Verde fluorescente.

质粒图谱

质粒序列

LOCUS       Exported                5073 bp ds-DNA     circular SYN 05-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 7
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5073)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, September 5, 2016 from SnapGene Viewer 3.1.4
FEATURES             Location/Qualifiers
     source          1..5073
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(115..703)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    1032..1079
                     /note="HSV TK poly(A) signal"
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(1311..2105)
                     /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"
     promoter        complement(2140..2497)
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      2154..2289
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(2499..2603)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      2630..3085
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     polyA_signal    3092..3213
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     CDS             complement(3745..4461)
                     /codon_start=1
                     /product="enhanced GFP"
                     /note="EGFP"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(4506..4709)
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        4710..5013
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atgcatggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
       61 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      121 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
      181 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
      241 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
      301 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
      361 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
      421 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
      481 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
      541 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
      601 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
      661 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
      721 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
      781 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
      841 ctaaagtata tatgagtaac ctgaggctat ggcagggcct gccgccccga cgttggctgc
      901 gagccctggg ccttcacccg aacttggggg gtggggtggg gaaaaggaag aaacgcgggc
      961 gtattggccc caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac
     1021 cccgcgttta tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt ttattgccgt
     1081 catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc tccccctagg
     1141 gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg
     1201 gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga
     1261 tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac
     1321 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
     1381 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
     1441 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
     1501 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
     1561 tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
     1621 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
     1681 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
     1741 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
     1801 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
     1861 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
     1921 tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
     1981 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     2041 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     2101 atcatgcgaa acgatcctca tcctgtctct tgatcgatct ttgcaaaagc ctaggcctcc
     2161 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     2221 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     2281 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     2341 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     2401 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     2461 ctgacacaca ttccacagct ggttctttcc gcctcaggac tcttcctttt tcaatattat
     2521 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2581 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     2641 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2701 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2761 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2821 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2881 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2941 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3001 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3061 aacaaaatat taacgcttac aatttacgcg ttaagataca ttgatgagtt tggacaaacc
     3121 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3181 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3241 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     3301 ggtatggctg attatgatca gttatctaga tccggtggat ccctacacag ccattgctgt
     3361 cccgaatgtc tcctgcgagg cataaaccat gtacaggaag ccgtcttcat ctctctcact
     3421 ctcgtacact tcggagatgg gagtggacac actcaccatg ctgtgcccat tcaccaggag
     3481 gaagaaggct tggttagcat tgagctgcaa gcgccgtctg attatcttga tgagctcgct
     3541 catgttcacg tggtcaggca ccaggaactt ggtcttgtcc aggacgggca gctgcttctc
     3601 ccccttgtat cgctctataa tcactgggat cttggtgggg tgctgctccc ggatgagccg
     3661 gacatcttcc actctttgtt caaagctccg gcgctgcttg aaggtcttct cggacggcat
     3721 aagcttgagc tcgagatctg agtacttgta cagctcgtcc atgccgagag tgatcccggc
     3781 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
     3841 ggcggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     3901 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat
     3961 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
     4021 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     4081 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     4141 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     4201 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc
     4261 gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     4321 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     4381 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     4441 ctcctcgccc ttgctcacca tggtggcgac cggtagcgct agcggatctg acggttcact
     4501 aaaccagctc tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat
     4561 ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc
     4621 ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc
     4681 ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg
     4741 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     4801 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     4861 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     4921 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     4981 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     5041 aatgaccccg taattgatta ctattaataa cta