pmTurquoise2-Golgi

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相关技术服务:质粒构建    基因合成    质粒大提

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载体基本信息

质粒名称: pmTurquoise2-Golgi
出品公司: Addgene
目录编号: 36205
存储实验室: Dorus Gadella

载体骨架基本信息
载体名称: pmTurquoise2-N1
载体出品公司: Clontech
空载体大小: 4904bp
完整质粒大小: 4904bp
载体类型: 哺乳细胞表达
筛选标记: 新霉素G418
载体抗性: Kanamycin
生长温度: 37 ℃
宿主菌: DH5a
拷贝数: 高拷贝

插入基因信息
插入基因名称: Golgi targeting sequence  /  B4GALT1(1-61) /beta4Gal-T1(1-61)/ beta-1,4-galactosyltransferase 1(1-61)
物种来源: H. sapiens (human)
插入大小: 183bp
GenBank编号 NM_001497
启动子: CMV
标签/融合蛋白: mTurquoise2 (C terminal on backbone)
克隆方法: 限制性内切酶
5’克隆位点: BglII (not destroyed)
3’克隆位点: AgeI (not destroyed)
5’测序引物 AGGTCTATATAAGCAGAGC
3’测序引物 TCTACAAATGTGGTATGGC

文献引用方法
材料和方法部分: pmTurquoise2-Golgi was a gift from Dorus Gadella (Addgene plasmid # 36205 ; http://n2t.net/addgene:36205 ; RRID:Addgene_36205)
参考文献部分: Structure-guided evolution of cyan fluorescent proteins towards a quantum yield of 93%. Goedhart J, von Stetten D, Noirclerc-Savoye M, Lelimousin M, Joosen L, Hink MA, van Weeren L, Gadella TW Jr, Royant A. Nat Commun. 2012 Mar 20;3:751. doi: 10.1038/ncomms1738. 10.1038/ncomms1738 PubMed 22434194

载体图谱质粒图谱和多克隆位点信息

pmTurquoise2-Golgi载体图谱

载体简介

载体序列

LOCUS       pmTurquoise2-Golgi                4904 bp ds-DNA     circular SYN 17-JUN-2022
DEFINITION  In vivo visualization of golgi apparatus (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    pmTurquoise2-Golgi
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4904)
  AUTHORS   Goedhart J, von Stetten D, Noirclerc-Savoye M, Lelimousin M, Joosen 
            L, Hink MA, van Weeren L, Gadella TW Jr, Royant A
  TITLE     Structure-guided evolution of cyan fluorescent proteins towards a 
            quantum yield of 93%.
  JOURNAL   Nat Commun. 2012 Mar 20;3:751. doi: 10.1038/ncomms1738.
  PUBMED    22434194
REFERENCE   2  (bases 1 to 4904)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2022 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..4904
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        9..312
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        313..516
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     467..487
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      792..801
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             798..1517
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(843..864)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1104..1123)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1451..1472
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1640..1761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1677..1696)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1731..1750
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1768..2223)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1905..1926)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2117..2136
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2250..2354
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2356..2713
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2379..2399)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2564..2699
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2626..2645
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2748..3542
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2802..2821)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3412..3431
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3730..3749)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3774..3821
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4150..4738
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4639..4658
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
       61 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      121 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      181 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      241 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      301 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      361 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      421 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      481 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      541 tagcgctacc ggactcagat ctcgagctcg ccaccatgag gcttcgggag ccgctcctga
      601 gcggcagcgc cgcgatgcca ggcgcgtccc tacagcgggc ctgccgcctg ctcgtggccg
      661 tctgcgctct gcaccttggc gtcaccctcg tttactacct ggctggccgc gacctgagcc
      721 gcctgcccca actggtcgga gtctccacac cgctgcagtc gacggtaccg cgggcccggg
      781 atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      841 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
      901 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
      961 ccgtgccctg gcccaccctc gtgaccaccc tgtcctgggg cgtgcagtgc ttcgcccgct
     1021 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1081 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1141 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1201 gcaacatcct ggggcacaag ctggagtaca actactttag cgacaacgtc tatatcaccg
     1261 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
     1321 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1381 tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga
     1441 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1501 acgagctgta caagtaaagc ggccgcgact ctagatcata atcagccata ccacatttgt
     1561 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     1621 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     1681 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     1741 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     1801 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     1861 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     1921 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     1981 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     2041 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2101 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     2161 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     2221 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2281 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2341 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     2401 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2461 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2521 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2581 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2641 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2701 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     2761 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     2821 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     2881 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     2941 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3001 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3061 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3121 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3181 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     3241 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     3301 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     3361 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     3421 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     3481 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     3541 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     3601 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     3661 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     3721 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     3781 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     3841 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     3901 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     3961 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     4021 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4081 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4141 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4201 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4261 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4321 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4381 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4441 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4501 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4561 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4621 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4681 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4741 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4801 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     4861 aatagtaatc aattacgggg tcattagttc atagcccata tatg
//

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