pCMV-tdTomato

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: pCMV-tdTomato
质粒类型: 哺乳动物细胞表达载体;荧光报告载体
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
载体大小: 5392 bp
5' 测序引物及序列: --
3' 测序引物及序列: --
载体标签: --
载体抗性: 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pCMV-tdTomato组成型表达tdTomato蛋白。
产品目录号: 632534
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

pCMV-tdTomato载体图谱
pCMV-tdTomato 载体特征

载体简介

pCMV-tdTomato载体描述

pCMV-tdTomato encodes tdTomato, a member of the family of fruit fluorescent proteins (1) derived from the Discosoma sp. red fluorescent protein, DsRed (2). Because the Tomato protein has a tendency to dimerize, the vector was designed with two copies of the Tomato coding region linked together to allow intramolecular dimerization. As a result, each tdTomato RNA transcript encodes a tandem dimer of the Tomato protein (excitation and emission maxima equal 554nm and 581nm, respectively). The tdTomato coding sequence is positioned just downstream of the constitutively active cytomegalovirus immediate early promoter (PCMV IE). As a result, mammalian cells transfected with this vector will constitutively express tdTomato. A Kozak consensus sequence is located immediately upstream of the tdTomato coding sequence to enhance translational efficiency in eukaryotic cells (3). SV40 polyadenylation signals downstream of the tdTomato coding sequence direct proper processing of the 3' end of the tdTomato mRNA. The vector backbone contains an SV40 origin for replication in mammalian cells expressing the SV40 large T antigen, a pUC origin of replication for propagation in E. coli, and an f1 origin for single-stranded DNA production. A neomycin resistance cassette (Neor) allows stably transfected eukaryotic cells to be selected using G418. This cassette consists of the SV40 early promoter, the neomycin/kanamycin resistance gene of Tn5, and polyadenylation signals from the Herpes simplex virus thymidine kinase (HSV TK) gene. A bacterial promoter upstream of the cassette expresses kanamycin resistance in E. coli.

pCMV-tdTomato is designed for use as a marker for cotransfection or cell tracking. The red fluorescence of tdTomato can be detected by fluorescence microscopy, allowing direct visual imaging. In addition, flow cytometry can be used to enrich transfected cell populations. pCMV-tdTomato can be transfected into mammalian cells using any standard transfection method. If required, stable transfectants can be selected using G418 (4).

Propagation in E. coli
 Suitable host strains: DH5α, HB101 and other general purpose strains. Single-stranded DNA production requires
a host containing an F plasmid such as JM109 or XL1-Blue.
 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/Col E1

Excitation and emission maxima of tdTomato
 Excitation maximum = 554 nm
 Emission maximum = 581 nm

References
1. Shaner, N. C., et al. (2004) Nature Biotech. 22(12):1567-1572.
2. Bevis, B. J. & Glick, B. S. (2002) Nature Biotech. 20:83–87.
3. Kozak, M. (1987) Nucleic Acids Res. 15:8125–8148.
4. Gorman, C. (1985) In DNA cloning: A Practical Approach, Vol. II. Ed. D. M. Glover. (IRL Press, Oxford, U.K.), pp. 143–190.


载体序列

LOCUS       pCMV-td-Tomato   5392 bp    DNA   circular            28-JUL-2008
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 632534
FEATURES             Location/Qualifiers
     misc_feature    3288..4082
                     /label=KanR/NeoR
     CDS             628..2055
                     /label=td-Tomato
     promoter        1..589
                     /label=pCMV\IE
BASE COUNT     1272 a      1505 c      1459 g      1156 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 gctagcgcta 
      601 ccggactcag atccaccggt cgccaccatg gtgagcaagg gcgaggaggt catcaaagag 
      661 ttcatgcgct tcaaggtgcg catggagggc tccatgaacg gccacgagtt cgagatcgag 
      721 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag 
      781 ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttcatgta cggctccaag 
      841 gcgtacgtga agcaccccgc cgacatcccc gattacaaga agctgtcctt ccccgagggc 
      901 ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt gacccaggac 
      961 tcctccctgc aggacggcac gctgatctac aaggtgaaga tgcgcggcac caacttcccc 
     1021 cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg 
     1081 tacccccgcg acggcgtgct gaagggcgag atccaccagg ccctgaagct gaaggacggc 
     1141 ggccactacc tggtggagtt caagaccatc tacatggcca agaagcccgt gcaactgccc 
     1201 ggctactact acgtggacac caagctggac atcacctccc acaacgagga ctacaccatc 
     1261 gtggaacagt acgagcgctc cgagggccgc caccacctgt tcctggggca tggcaccggc 
     1321 agcaccggca gcggcagctc cggcaccgcc tcctccgagg acaacaacat ggccgtcatc 
     1381 aaagagttca tgcgcttcaa ggtgcgcatg gagggctcca tgaacggcca cgagttcgag 
     1441 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg 
     1501 accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt catgtacggc 
     1561 tccaaggcgt acgtgaagca ccccgccgac atccccgatt acaagaagct gtccttcccc 
     1621 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc 
     1681 caggactcct ccctgcagga cggcacgctg atctacaagg tgaagatgcg cggcaccaac 
     1741 ttcccccccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag 
     1801 cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc accaggccct gaagctgaag 
     1861 gacggcggcc actacctggt ggagttcaag accatctaca tggccaagaa gcccgtgcaa 
     1921 ctgcccggct actactacgt ggacaccaag ctggacatca cctcccacaa cgaggactac 
     1981 accatcgtgg aacagtacga gcgctccgag ggccgccacc acctgttcct gtacggcatg 
     2041 gacgagctgt acaagtaggc ggccgcgact ctagatcata atcagccata ccacatttgt 
     2101 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat 
     2161 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa 
     2221 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 
     2281 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg 
     2341 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc 
     2401 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga 
     2461 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg 
     2521 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag 
     2581 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga 
     2641 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg 
     2701 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg 
     2761 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 
     2821 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 
     2881 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg 
     2941 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 
     3001 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 
     3061 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc 
     3121 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc 
     3181 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct 
     3241 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag 
     3301 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg 
     3361 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc 
     3421 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag 
     3481 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca 
     3541 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat 
     3601 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata 
     3661 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac 
     3721 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc 
     3781 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg 
     3841 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg 
     3901 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta 
     3961 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg 
     4021 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct 
     4081 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga 
     4141 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc 
     4201 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg 
     4261 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat 
     4321 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc 
     4381 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt 
     4441 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa 
     4501 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt 
     4561 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac 
     4621 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 
     4681 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 
     4741 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 
     4801 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg 
     4861 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 
     4921 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 
     4981 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 
     5041 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 
     5101 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 
     5161 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 
     5221 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 
     5281 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt 
     5341 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc at 
//

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