pAcGFP1-Mito

价格:2300元

联系方式:I47-825O-882O

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载体基本信息

出品公司: Clontech
载体名称: pAcGFP1-Mito
质粒类型: 哺乳动物细胞表达载体;荧光报告载体;亚细胞定位载体
高拷贝/低拷贝: 高拷贝
克隆方法: 限制性内切酶,多克隆位点
启动子: CMV IE
载体大小: 4.8 kb
5' 测序引物及序列: --
3' 测序引物及序列: --
载体标签: --
载体抗性: Kanamycin.html' target='_blank'>卡那霉素
筛选标记: 新霉素(Neomycin)
克隆菌株: DH5α, HB101
宿主细胞(系): 常规细胞系(293、CV-1、CHO等)
备注: 哺乳动物载体pAcGFP1-Mito组成型表达AcGFP,是线粒体定位载体。
产品目录号: 632432
稳定性: 稳表达 或 瞬表达
组成型/诱导型: 组成型
病毒/非病毒: 非病毒

载体质粒图谱和多克隆位点信息

pAcGFP1-Mito载体图谱



pAcGFP1-Mito 载体特征

载体简介

pAcGFP1-Mito encodes a fusion of a mitochondrial targeting sequence derived from the precursor of subunit VIII of human cytochrome C oxidase (1, 2) and the green fluorescent protein (GFP) from Aequorea coerulescens (AcGFP1; excitation maximum = 475 nm; emission maximum = 505 nm). The mitochondrial targeting sequence is fused to the N-terminus of AcGFP1. AcGFP1 contains silent mutations that create an open reading frame comprised almost entirely of optimized human codons. These changes increase the translational efficiency of the AcGFP1 mRNA and, consequently, the expression of AcGFP1 in mammalian and plant cells. The vector contains an SV40 origin for replication and a neomycin resistance (Neor) gene for selection (using G418) in eukaryotic cells (3). A bacterial promoter (P) upstream of Neor expresses kanamycin resistance in E. coli. The vector backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.

pAcGFP1-Mito is designed for fluorescent labeling of mitochondria. The fluorescence from pAcGFP1-Mito expression can be observed within the mitochondrial matrix inside the inner membrane. pAcGFP1-Mito can be introduced into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418 (3). pAcGFP1- Mito is not intended as a cloning vector; however, the backbone does contain unique restriction sites upstream and downstream of the AcGFP1-Mito sequence which permit excision of the AcGFP1-Mito sequence.

Propagation in E. coli
  Suitable host strains: DH5α, HB101, and other general purpose strains. Single-stranded DNA production requires a host such as JM109 or XL1-Blue that contains an F plasmid .
  Selectable marker: plasmid confers resistance to kanamycin (50 μg/ml) to E. coli hosts.
  E. coli replication origin: pUC
  Copy number: ≈500
  Plasmid incompatibility group: pMB1/ColE1

载体序列

LOCUS       pAcGFP1-Mito   4752 bp    DNA   circular            15-JAN-2010
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 632432
FEATURES             Location/Qualifiers
     misc_feature    1..589
                     /label=CMV\IE\Promoter
     misc_feature    699..1418
                     /label=AcGFP1
     misc_feature    597..683
                     /label=Mitochondrial\targeting\sequence
     misc_feature    597..1418
                     /label=AcGFP1-Mito\fusion
     misc_feature    1571..1576
                     /label=SV40\early\polyA\signal
     misc_feature    1600..1605
                     /label=SV40\early\polyA\signal
     misc_feature    1669..2123
                     /label=f1\origin
     misc_feature    2185..2213
                     /label=Bacterial\promoter\for\Kan(R)
     misc_feature    2297..2526
                     /label=SV40\early\Promoter/Enhancer
     misc_feature    2464..2599
                     /label=SV40\origin
     misc_feature    2548..3442
                     /label=Kan(R)/Neo(R)
     misc_feature    3678..3683
                     /label=HSV\TK\polyA\signal
     misc_feature    3691..3696
                     /label=HSV\TK\polyA\signal
     misc_feature    4027..4670
                     /label=pUC\origin
BASE COUNT     1135 a      1278 c      1253 g      1086 t 
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 gctagcatgt 
      601 ccgtcctgac gccgctgctg ctgcggggct tgacaggctc ggcccggcgg ctcccagtgc 
      661 cgcgcgccaa gatccattcg ttgggggatc caccggtcat ggtgagcaag ggcgccgagc 
      721 tgttcaccgg catcgtgccc atcctgatcg agctgaatgg cgatgtgaat ggccacaagt 
      781 tcagcgtgag cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca 
      841 tctgcaccac cggcaagctg cctgtgccct ggcccaccct ggtgaccacc ctgagctacg 
      901 gcgtgcagtg cttctcacgc taccccgatc acatgaagca gcacgacttc ttcaagagcg 
      961 ccatgcctga gggctacatc caggagcgca ccatcttctt cgaggatgac ggcaactaca 
     1021 agtcgcgcgc cgaggtgaag ttcgagggcg ataccctggt gaatcgcatc gagctgaccg 
     1081 gcaccgattt caaggaggat ggcaacatcc tgggcaataa gatggagtac aactacaacg 
     1141 cccacaatgt gtacatcatg accgacaagg ccaagaatgg catcaaggtg aacttcaaga 
     1201 tccgccacaa catcgaggat ggcagcgtgc agctggccga ccactaccag cagaataccc 
     1261 ccatcggcga tggccctgtg ctgctgcccg ataaccacta cctgtccacc cagagcgccc 
     1321 tgtccaagga ccccaacgag aagcgcgatc acatgatcta cttcggcttc gtgaccgccg 
     1381 ccgccatcac ccacggcatg gatgagctgt acaagtgagc ggccgcgact ctagatcata 
     1441 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc 
     1501 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat 
     1561 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 
     1621 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt 
     1681 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata 
     1741 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt 
     1801 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg 
     1861 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt 
     1921 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc 
     1981 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg 
     2041 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct 
     2101 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc 
     2161 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg 
     2221 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg 
     2281 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 
     2341 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 
     2401 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 
     2461 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 
     2521 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga 
     2581 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc 
     2641 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 
     2701 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 
     2761 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 
     2821 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 
     2881 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 
     2941 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 
     3001 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 
     3061 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 
     3121 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat 
     3181 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 
     3241 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 
     3301 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 
     3361 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 
     3421 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg 
     3481 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc 
     3541 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag 
     3601 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga 
     3661 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt 
     3721 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat 
     3781 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa 
     3841 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca 
     3901 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc 
     3961 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca 
     4021 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 
     4081 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 
     4141 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt 
     4201 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 
     4261 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 
     4321 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 
     4381 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 
     4441 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 
     4501 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 
     4561 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 
     4621 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 
     4681 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 
     4741 accgccatgc at 
//

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