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ptdTomato载体

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  • 中国/美国
  • ZY2300
  • 2025年07月15日
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    • 详细信息
    • 文献和实验
    • 技术资料
    • 保存条件

      -20℃低温保存

    • 保质期

      三年

    • 英文名

      ptdTomato

    • 库存

      20

    • 供应商

      泽叶生物

    载体基本信息

    出品公司: Clontech
    载体名称: ptdTomato
    质粒类型: 原核表达载体;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: lac
    载体大小: 4065 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: 氨苄青霉素
    筛选标记: --
    克隆菌株: DH5α
    宿主细胞(系): 大肠杆菌
    备注: 原核表达载体ptdTomato在IPTG诱导下表达tdTomato红色荧光蛋白;也可作为tdTomato基因的克隆载体。
    产品目录号: ZY632531
    稳定性: 稳表达
    组成型/诱导型: 诱导型
    病毒/非病毒: 非病毒

    载体质粒图谱和多克隆位点信息

    ptdTomato载体图谱



    ptdTomato 多克隆位点

    ptdTomato 载体特征

    载体简介

    ptdTomato载体描述
    
    ptdTomato 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 desighed with two copies of the Tomato coding region liked 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). In ptdTomato, the tdTomato coding sequence (2) is flanked by restriction sites (5' MCS and 3' MCS) that make it easy to excise the gene for use in other cloning applications. In E. coli, tdTomato is expressed from the lac promoter (Plac) as a fusion with several amino acids, including the first five amino acids of the LacZ protein. Note, however, that if the tdTomato coding sequence is excised using a restriction site in the 5' MCS, the protein will no longer be expressed as a fusion (as it is when expressed from the lac promoter).A Kozak consensus sequence is located immediately upstream of the tdTomato coding sequence to enhance translational efficiency in eukaryotic cells (3). In the tdTomato vector, the entire tdTomato expression cassette is supported by a pUC19 backbone, which contains a high copy-number origin of replication and an ampicillin resistance gene for propagation and selection in E. coli.
    
    The ptdTomato vector is primarily intended to serve as a source of tdTomato cDNA. The flanking MCS regions make it possible to excise the tdTomato coding sequence and insert it into other vector systems. The vector can also be used to express tdTomato in bacteria.
    For Western analysis, either the Living Colors DsRed Polyclonal Antibody (Cat. No. 632496) or DsRed Monoclonal Antibody (Cat. Nos. 632392 and 632393) can be used to detect the tdTomato protein.
    
    Propagation in E. Coli
     Recommended host strain: DH5α
     Selectable marker: plasmid confers resistance to ampicillin (50 μg/ml) in E. coli hosts.
     E. coli replication origin: pUC
     Copy number: high
     Plasmid incompatibility group: pMB1/ColE1
    
    Excitation and emission maxima of tdTomato
     Excitation maximum = 554 nm
     Emission maximum = 581 nm
    
    

    载体序列

    LOCUS       ptdTomato	4065 bp 	DNA	SYN
    DEFINITION  ptdTomato
    ACCESSION   
    KEYWORDS    
    SOURCE      
      ORGANISM  other sequences; artificial sequences; vectors.
    FEATURES             Location/Qualifiers
         source          1..4065
                         /organism="ptdTomato"
                         /mol_type="other DNA"
         promoter        143..172
                         /label="lac_promoter"
         misc_feature    186..208
                         /label="M13_pUC_rev_primer"
         promoter        207..225
                         /label="M13_reverse_primer"
         CDS             217..1719
                         /label="ORF frame 1"
                         /translation="MTMITPSLHACRSTLEDPRVPVATMVSKGEEVIKEFMRFKVRME
                         GSMNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPA
                         DIPDYKKLSFPEGFKWERVMNFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPV
                         MQKKTMGWEASTERLYPRDGVLKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYY
                         YVDTKLDITSHNEDYTIVEQYERSEGRHHLFLGHGTGSTGSGSSGTASSEDNNMAVIK
                         EFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMY
                         GSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGLVTVTQDSSLQDGTLIYKVKMR
                         GTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQALKLKDGGHYLVEFKTIYMA
                         KKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHHLFLYGMDELYK*"
         CDS             complement(259..972)
                         /label="ORF frame 1"
                         /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL
                         GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA
                         AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA
                         VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL
                         FDDLLALAHHGGDRYPGIL*"
         gene            292..1716
                         /label="dTomato (variant)"
                         /gene="dTomato (variant)"
                         /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL
                         GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA
                         AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA
                         VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL
                         FDDLLALAHHGGDRYPGIL*"
         misc_feature    457..510
                         /label="Q66M (mCherry)"
                         /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL
                         GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA
                         AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA
                         VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL
                         FDDLLALAHHGGDRYPGIL*"
         misc_feature    884..907
                         /label="dsRed1_C_primer"
                         /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL
                         GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA
                         AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA
                         VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL
                         FDDLLALAHHGGDRYPGIL*"
         misc_feature    1610..1633
                         /label="dsRed1_C_primer"
                         /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL
                         GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA
                         AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA
                         VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL
                         FDDLLALAHHGGDRYPGIL*"
         misc_feature    complement(1855..1877)
                         /label="pGEX_3_primer"
                         /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL
                         GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA
                         AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA
                         VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL
                         FDDLLALAHHGGDRYPGIL*"
         promoter        2195..2223
                         /label="AmpR_promoter"
                         /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL
                         GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA
                         AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA
                         VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL
                         FDDLLALAHHGGDRYPGIL*"
         gene            2265..3125
                         /label="Ampicillin"
                         /gene="Ampicillin"
                         /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL
                         GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA
                         AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA
                         VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL
                         FDDLLALAHHGGDRYPGIL*"
         CDS             2265..3125
                         /label="ORF frame 3"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
         rep_origin      3280..3899
                         /label="pBR322_origin"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
    ORIGIN
        1 AGCGCCCAAT ACGCAAACCG CCTCTCCCCG CGCGTTGGCC GATTCATTAA TGCAGCTGGC
       61 ACGACAGGTT TCCCGACTGG AAAGCGGGCA GTGAGCGCAA CGCAATTAAT GTGAGTTAGC
      121 TCACTCATTA GGCACCCCAG GCTTTACACT TTATGCTTCC GGCTCGTATG TTGTGTGGAA
      181 TTGTGAGCGG ATAACAATTT CACACAGGAA ACAGCTATGA CCATGATTAC GCCAAGCTTG
      241 CATGCCTGCA GGTCGACTCT AGAGGATCCC CGGGTACCGG TCGCCACCAT GGTGAGCAAG
      301 GGCGAGGAGG TCATCAAAGA GTTCATGCGC TTCAAGGTGC GCATGGAGGG CTCCATGAAC
      361 GGCCACGAGT TCGAGATCGA GGGCGAGGGC GAGGGCCGCC CCTACGAGGG CACCCAGACC
      421 GCCAAGCTGA AGGTGACCAA GGGCGGCCCC CTGCCCTTCG CCTGGGACAT CCTGTCCCCC
      481 CAGTTCATGT ACGGCTCCAA GGCGTACGTG AAGCACCCCG CCGACATCCC CGATTACAAG
      541 AAGCTGTCCT TCCCCGAGGG CTTCAAGTGG GAGCGCGTGA TGAACTTCGA GGACGGCGGT
      601 CTGGTGACCG TGACCCAGGA CTCCTCCCTG CAGGACGGCA CGCTGATCTA CAAGGTGAAG
      661 ATGCGCGGCA CCAACTTCCC CCCCGACGGC CCCGTAATGC AGAAGAAGAC CATGGGCTGG
      721 GAGGCCTCCA CCGAGCGCCT GTACCCCCGC GACGGCGTGC TGAAGGGCGA GATCCACCAG
      781 GCCCTGAAGC TGAAGGACGG CGGCCACTAC CTGGTGGAGT TCAAGACCAT CTACATGGCC
      841 AAGAAGCCCG TGCAACTGCC CGGCTACTAC TACGTGGACA CCAAGCTGGA CATCACCTCC
      901 CACAACGAGG ACTACACCAT CGTGGAACAG TACGAGCGCT CCGAGGGCCG CCACCACCTG
      961 TTCCTGGGGC ATGGCACCGG CAGCACCGGC AGCGGCAGCT CCGGCACCGC CTCCTCCGAG
     1021 GACAACAACA TGGCCGTCAT CAAAGAGTTC ATGCGCTTCA AGGTGCGCAT GGAGGGCTCC
     1081 ATGAACGGCC ACGAGTTCGA GATCGAGGGC GAGGGCGAGG GCCGCCCCTA CGAGGGCACC
     1141 CAGACCGCCA AGCTGAAGGT GACCAAGGGC GGCCCCCTGC CCTTCGCCTG GGACATCCTG
     1201 TCCCCCCAGT TCATGTACGG CTCCAAGGCG TACGTGAAGC ACCCCGCCGA CATCCCCGAT
     1261 TACAAGAAGC TGTCCTTCCC CGAGGGCTTC AAGTGGGAGC GCGTGATGAA CTTCGAGGAC
     1321 GGCGGTCTGG TGACCGTGAC CCAGGACTCC TCCCTGCAGG ACGGCACGCT GATCTACAAG
     1381 GTGAAGATGC GCGGCACCAA CTTCCCCCCC GACGGCCCCG TAATGCAGAA GAAGACCATG
     1441 GGCTGGGAGG CCTCCACCGA GCGCCTGTAC CCCCGCGACG GCGTGCTGAA GGGCGAGATC
     1501 CACCAGGCCC TGAAGCTGAA GGACGGCGGC CACTACCTGG TGGAGTTCAA GACCATCTAC
     1561 ATGGCCAAGA AGCCCGTGCA ACTGCCCGGC TACTACTACG TGGACACCAA GCTGGACATC
     1621 ACCTCCCACA ACGAGGACTA CACCATCGTG GAACAGTACG AGCGCTCCGA GGGCCGCCAC
     1681 CACCTGTTCC TGTACGGCAT GGACGAGCTG TACAAGTAGG CGGCCGCGAC TCTAGAATTC
     1741 CAACTGAGCG CCGGTCGCTA CCATTACCAA CTTGTCTGGT GTCAAAAATA ATAGGCCTAC
     1801 TAGTCGGCCG TACGGGCCCT TTCGTCTCGC GCGTTTCGGT GATGACGGTG AAAACCTCTG
     1861 ACACATGCAG CTCCCGGAGA CGGTCACAGC TTGTCTGTAA GCGGATGCCG GGAGCAGACA
     1921 AGCCCGTCAG GGCGCGTCAG CGGGTGTTGG CGGGTGTCGG GGCTGGCTTA ACTATGCGGC
     1981 ATCAGAGCAG ATTGTACTGA GAGTGCACCA TATGCGGTGT GAAATACCGC ACAGATGCGT
     2041 AAGGAGAAAA TACCGCATCA GGCGGCCTTA AGGGCCTCGT GATACGCCTA TTTTTATAGG
     2101 TTAATGTCAT GATAATAATG GTTTCTTAGA CGTCAGGTGG CACTTTTCGG GGAAATGTGC
     2161 GCGGAACCCC TATTTGTTTA TTTTTCTAAA TACATTCAAA TATGTATCCG CTCATGAGAC
     2221 AATAACCCTG ATAAATGCTT CAATAATATT GAAAAAGGAA GAGTATGAGT ATTCAACATT
     2281 TCCGTGTCGC CCTTATTCCC TTTTTTGCGG CATTTTGCCT TCCTGTTTTT GCTCACCCAG
     2341 AAACGCTGGT GAAAGTAAAA GATGCTGAAG ATCAGTTGGG TGCACGAGTG GGTTACATCG
     2401 AACTGGATCT CAACAGCGGT AAGATCCTTG AGAGTTTTCG CCCCGAAGAA CGTTTTCCAA
     2461 TGATGAGCAC TTTTAAAGTT CTGCTATGTG GCGCGGTATT ATCCCGTATT GACGCCGGGC
     2521 AAGAGCAACT CGGTCGCCGC ATACACTATT CTCAGAATGA CTTGGTTGAG TACTCACCAG
     2581 TCACAGAAAA GCATCTTACG GATGGCATGA CAGTAAGAGA ATTATGCAGT GCTGCCATAA
     2641 CCATGAGTGA TAACACTGCG GCCAACTTAC TTCTGACAAC GATCGGAGGA CCGAAGGAGC
     2701 TAACCGCTTT TTTGCACAAC ATGGGGGATC ATGTAACTCG CCTTGATCGT TGGGAACCGG
     2761 AGCTGAATGA AGCCATACCA AACGACGAGC GTGACACCAC GATGCCTGTA GCAATGGCAA
     2821 CAACGTTGCG CAAACTATTA ACTGGCGAAC TACTTACTCT AGCTTCCCGG CAACAATTAA
     2881 TAGACTGGAT GGAGGCGGAT AAAGTTGCAG GACCACTTCT GCGCTCGGCC CTTCCGGCTG
     2941 GCTGGTTTAT TGCTGATAAA TCTGGAGCCG GTGAGCGTGG GTCTCGCGGT ATCATTGCAG
     3001 CACTGGGGCC AGATGGTAAG CCCTCCCGTA TCGTAGTTAT CTACACGACG GGGAGTCAGG
     3061 CAACTATGGA TGAACGAAAT AGACAGATCG CTGAGATAGG TGCCTCACTG ATTAAGCATT
     3121 GGTAACTGTC AGACCAAGTT TACTCATATA TACTTTAGAT TGATTTAAAA CTTCATTTTT
     3181 AATTTAAAAG GATCTAGGTG AAGATCCTTT TTGATAATCT CATGACCAAA ATCCCTTAAC
     3241 GTGAGTTTTC GTTCCACTGA GCGTCAGACC CCGTAGAAAA GATCAAAGGA TCTTCTTGAG
     3301 ATCCTTTTTT TCTGCGCGTA ATCTGCTGCT TGCAAACAAA AAAACCACCG CTACCAGCGG
     3361 TGGTTTGTTT GCCGGATCAA GAGCTACCAA CTCTTTTTCC GAAGGTAACT GGCTTCAGCA
     3421 GAGCGCAGAT ACCAAATACT GTTCTTCTAG TGTAGCCGTA GTTAGGCCAC CACTTCAAGA
     3481 ACTCTGTAGC ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG GCTGCTGCCA
     3541 GTGGCGATAA GTCGTGTCTT ACCGGGTTGG ACTCAAGACG ATAGTTACCG GATAAGGCGC
     3601 AGCGGTCGGG CTGAACGGGG GGTTCGTGCA CACAGCCCAG CTTGGAGCGA ACGACCTACA
     3661 CCGAACTGAG ATACCTACAG CGTGAGCTAT GAGAAAGCGC CACGCTTCCC GAAGGGAGAA
     3721 AGGCGGACAG GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG AGGGAGCTTC
     3781 CAGGGGGAAA CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC TGACTTGAGC
     3841 GTCGATTTTT GTGATGCTCG TCAGGGGGGC GGAGCCTATG GAAAAACGCC AGCAACGCGG
     3901 CCTTTTTACG GTTCCTGGCC TTTTGCTGGC CTTTTGCTCA CATGTTCTTT CCTGCGTTAT
     3961 CCCCTGATTC TGTGGATAAC CGTATTACCG CCTTTGAGTG AGCTGATACC GCTCGCCGCA
     4021 GCCGAACGAC CGAGCGCAGC GAGTCAGTGA GCGAGGAAGC GGAAG

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