• 商品货号：FH1594

• 规格：10μl质粒
• 启动子：CMV promoter
• 复制子：pUC ori,f1 ori
• 终止子：SV40 poly(A) signal
• 质粒大小：5442bp
• 质粒标签：N-tdTomato
• 原核抗性：卡那霉素Kan
• 筛选标记：新霉素Neo/G418
• 克隆菌株：大肠杆菌Stbl3
• 表达宿主：哺乳细胞293T
• 诱导方式：无须诱导，瞬时表达
• 5测序引物：根据序列设计引物
• 3测序引物：Sv40-polyA-R：GAAATTTGTGATGCTATTGC
• 质粒简介：          ptdTomato-C1质粒是一个哺乳细胞红色荧光过表达质粒，CMV启动子驱动tdTomato红色荧光蛋白和目的基因融合在一起表达。
          ptdTomato-C1 is a mammalian expression vector designed to express a protein of interest fused to the C-terminus of tdTomato. td Tomato is 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). Expression of td Tomato as a tandem dimer prevents the fused protein of interest from being forced into a dimeric complex. Fusions that retain the fluorescence properties of tdTomato can be monitored by flow cytometry and localized by fluorescence microscopy. The multiple cloning site (MCS) in ptdTomato-C1 is positioned downstream of the tdTomato coding sequence. A Kozak consensus sequence, located immediately upstream of the tdTomato coding sequence, enhances translational efficiency in eukaryotic cells (3). SV40 polyadenylation signals downstream of the tdTomato coding sequence and the MCS direct proper processing of the 3' end of the tdTomato (or fusion gene) 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. This vector also has a neomycin resistance cassette (Neor) that allows G418 selection of stably transfected eukaryotic cells. This cassette consists of the SV40 early promoter, the Tn5 neomycin/kanamycin resistance gene, and polyadenylation signals from the herpes simplex virus thymidine kinase (HSV TK) gene. A bacterial promoter upstream of the cassette confers kanamycin resistance in E. Coli.
• 质粒图谱1： png/4-160Z1160522O1.png

质粒序列： 点击查看序列

LOCUS       Exported                5442 bp ds-DNA     circular SYN 01-SEP-2016

DEFINITION  synthetic circular DNA

KEYWORDS    Untitled 8

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 5442)

  TITLE     Direct Submission

            http://www.fenghbio.cn

FEATURES             Location/Qualifiers

     source          1..5442

                     /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"

     CDS             613..2040

                     /codon_start=1

                     /product="tandem dimeric (pseudo-monomeric) derivative of 

                     DsRed (Shaner et al., 2004)"

                     /note="tdTomato"

                     /note="mammalian codon-optimized"

                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA

                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG

                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH

                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH

                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY

                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM

                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV

                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE

                     RSEGRHHLFLYGMDELYK"

     misc_feature    2041..2106

                     /note="MCS"

                     /note="multiple cloning site"

     polyA_signal    2230..2351

                     /note="SV40 poly(A) signal"

                     /note="SV40 polyadenylation signal"

     rep_origin      complement(2358..2813)

                     /direction=LEFT

                     /note="f1 ori"

                     /note="f1 bacteriophage origin of replication; arrow 

                     indicates direction of (+) strand synthesis"

     promoter        2840..2944

                     /gene="bla"

                     /note="AmpR promoter"

     promoter        2946..3303

                     /note="SV40 promoter"

                     /note="SV40 enhancer and early promoter"

     rep_origin      3154..3289

                     /note="SV40 ori"

                     /note="SV40 origin of replication"

     CDS             3338..4132

                     /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    4364..4411

                     /note="HSV TK poly(A) signal"

                     /note="herpes simplex virus thymidine kinase 

                     polyadenylation signal (Cole and Stacy, 1985)"

     rep_origin      4740..5328

                     /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 ccggtcgcca ccatggtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag

      661 gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc

      721 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc

      781 ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac

      841 cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc

      901 gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac

      961 ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta

     1021 atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc

     1081 gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg

     1141 gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg

     1201 gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag

     1261 cgctccgagg gccgccacca cctgttcctg gggcatggca ccggcagcac cggcagcggc

     1321 agctccggca ccgcctcctc cgaggacaac aacatggccg tcatcaaaga gttcatgcgc

     1381 ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc

     1441 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc

     1501 ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg

     1561 aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg

     1621 gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg

     1681 caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc

     1741 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc

     1801 gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac

     1861 ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac

     1921 tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag

     1981 tacgagcgct ccgagggccg ccaccacctg ttcctgtacg gcatggacga gctgtacaag

     2041 tccggactca gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg

     2101 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta

     2161 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt

     2221 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca

     2281 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc

     2341 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt

     2401 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc

     2461 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt

     2521 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact

     2581 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg

     2641 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag

     2701 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac

     2761 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg

     2821 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa

     2881 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa

     2941 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc

     3001 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg

     3061 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag

     3121 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc

     3181 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc

     3241 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc

     3301 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca

     3361 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac

     3421 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt

     3481 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc

     3541 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg

     3601 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc

     3661 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc

     3721 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat

     3781 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc

     3841 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca

     3901 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga

     3961 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat

     4021 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc

     4081 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact

     4141 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc

     4201 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg

     4261 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact

     4321 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag

     4381 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg

     4441 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt

     4501 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg

     4561 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat

     4621 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct

     4681 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct