pIRES2-DsRed-Express

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P0150 pIRES2-DsRed-Express 質粒干粉 QQ詢價 QQ詢價
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啟動子: CMV
復制子: pUC
終止子: SV40 poly(A) signal
質粒分類: 哺乳細胞,熒光蛋白報告載體
質粒大小: 5264bp
原核抗性: Kan
真核抗性: G418
克隆菌株: DH5a
培養條件: 37度
表達宿主: 哺乳細胞
誘導方式: 無須誘導,瞬時表達
5'測序引物: CMV-F:CGCAAATGGGCGGTAGGCGTG
3'測序引物: 根據序列設計引物
質粒宿主: 哺乳細胞
質粒用途: 蛋白表達
片段類型: ORF
片段物種: 空載體
原核抗性: Kan
真核抗性: G418
熒光標記: 紅色
哺乳細胞表達空載體
     pIRES2-DsRed-Express contains the internal ribosome entry site (IRES; 1, 2) of the encephalomyocarditis virus (ECMV) between the MCS and a mutant of the Discosoma sp. red fluorescent protein DsRed-Express coding region (3–5). This permits both the gene of interest (cloned into the MCS) and the DsRed-Express gene to be translated from a single bicistronic mRNA. pIRES2-DsRed-Express is designed for the efficient selection by flow cytometry of transiently transfected mammalian cells expressing DsRed-Express and the protein of interest. This vector can also be used to express DsRed-Express alone or to obtain stably transfected cell lines without timeconsuming drug and clonal selection.
LOCUS       Exported                5264 bp ds-DNA     circular SYN 01-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 7
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5264)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, September 1, 2016 from SnapGene Viewer 3.1.4
            http://www.9687716.buzz
FEATURES             Location/Qualifiers
     source          1..5264
                     /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    609..665
                     /note="MCS"
                     /note="multiple cloning site"
     misc_feature    667..1253
                     /note="IRES2"
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             1254..1931
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /note="DsRed-Express"
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     polyA_signal    2052..2173
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2180..2635)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2662..2766
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        2768..3125
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      2976..3111
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             3160..3954
                     /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    4186..4233
                     /note="HSV TK poly(A) signal"
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4562..5150
                     /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 gatccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg
      721 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
      781 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
      841 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
      901 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
      961 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
     1021 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
     1081 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
     1141 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
     1201 ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggcct
     1261 cctccgagga cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga
     1321 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     1381 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc
     1441 cccagttcca gtacggctcc aaggtgtacg tgaagcaccc cgccgacatc cccgactaca
     1501 agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     1561 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg ctccttcatc tacaaggtga
     1621 agttcatcgg cgtgaacttc ccctccgacg gccccgtaat gcagaagaag actatgggct
     1681 gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccaca
     1741 aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagtct atctatatgg
     1801 ccaagaagcc cgtgcagctg cccggctact actacgtgga ctccaagctg gacatcacct
     1861 cccacaacga ggactacacc atcgtggagc agtacgagcg cgccgagggc cgccaccacc
     1921 tgttcctgta gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     1981 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2041 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2101 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2161 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     2221 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     2281 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     2341 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2401 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     2461 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     2521 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     2581 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     2641 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2701 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2761 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2821 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2881 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2941 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3001 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3061 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3121 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3181 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3241 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3301 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     3361 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3421 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3481 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3541 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3601 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3661 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     3721 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     3781 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3841 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     3901 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     3961 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4021 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4081 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     4141 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     4201 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     4261 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     4321 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     4381 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     4441 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4501 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4561 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4621 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4681 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4741 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4801 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4861 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4921 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4981 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5041 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5101 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5161 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5221 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
 
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