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基本信息

質粒圖譜和多克隆位點信息

注意：

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如要使用本公司保存的母源載體時，本公司不能保證所有母源載體完全符合客戶預期，也不承擔由此產生的任何責任。

載體描述

pCMV-C-mCherry為哺乳動物細胞表達質粒，用于表達C端含mCherry (DsRed的突變體，一種非常明亮的紅色熒光蛋白)標簽的融合蛋白。該質粒含有CMV啟動子，可以高效啟動目的蛋白在細胞中的表達。在多克隆位點的后面有一個mCherry的完整編碼序列，因此在多克隆位點根據閱讀框插入目的基因就可以表達C端含有mCherry標簽的融合蛋白。利用mCherry的熒光特性可以比較容易地觀察融合蛋白的表達水平和細胞內定位，也可以利用mCherry抗體來檢測或免疫沉淀融合蛋白。mCherry與GFP沒有序列同源性，不能使用GFP抗體檢測mCherry。該質粒為卡那霉素抗性。轉染細胞后，可以使用G418篩選穩定表達目的蛋白的細胞株。

載體序列

LOCUS       Exported                5001 bp ds-DNA     circular SYN 20-MAR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    .

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 5001)

  AUTHORS   BT-Lab

  TITLE     Direct Submission

  JOURNAL   Exported Wednesday, Mar 20, 2019 from SnapGene Viewer 4.3.4

FEATURES             Location/Qualifiers

     source          1..5001

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     enhancer        67..370

                     /label=CMV enhancer

                     /note="human cytomegalovirus immediate early enhancer"

     promoter        371..574

                     /label=CMV promoter

                     /note="human cytomegalovirus (CMV) immediate early

                     promoter"

     primer_bind     525..545

                     /label=CMV F

     promoter        620..638

                     /label=T3 promoter

                     /note="promoter for bacteriophage T3 RNA polymerase"

     misc_feature    651..740

                     /label=MCS

                     /note="Multiple cloning site"

     CDS             741..1451

                     /codon_start=1

                     /product="monomeric derivative of DsRed fluorescent protein

                     (Shaner et al., 2004)"

                     /label=mCherry

                     /note="mammalian codon-optimized"

                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG

                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF

                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK

                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA

                     EGRHSTGGMDELYK"

     promoter        complement(1503..1521)

                     /label=T7 promoter

                     /note="promoter for bacteriophage T7 RNA polymerase"

     polyA_signal    1795..1916

                     /label=SV40 poly(A) signal

                     /note="SV40 polyadenylation signal"

     rep_origin      complement(1923..2378)

                     /direction=LEFT

                     /label=f1 ori

                     /note="f1 bacteriophage origin of replication; arrow

                     indicates direction of (+) strand synthesis"

     promoter        2405..2509

                     /gene="bla"

                     /label=AmpR promoter

     promoter        2511..2868

                     /label=SV40 promoter

                     /note="SV40 enhancer and early promoter"

     rep_origin      2719..2854

                     /label=SV40 ori

                     /note="SV40 origin of replication"

     CDS             2903..3697

                     /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"

     polyA_signal    3929..3976

                     /label=HSV TK poly(A) signal

                     /note="herpes simplex virus thymidine kinase

                     polyadenylation signal (Cole and Stacy, 1985)"

     rep_origin      4305..4893

                     /direction=RIGHT

                     /label=ori

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of

                     replication"

ORIGIN

        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga

       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg

      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg

      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca

      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc

      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc

      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc

      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa

      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag

      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta

      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc

      661 gcggtggcgg ccgctctagc ccgggcggat ccaagcttct gcaggaattc gatatcgtcg

      721 acagatctct cgagtctaga atggtgagca agggcgagga ggataacatg gccatcatca

      781 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga

      841 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga

      901 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct

      961 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg

     1021 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc

     1081 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact

     1141 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc

     1201 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg

     1261 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc

     1321 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca

     1381 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc

     1441 tgtacaagta ggggcccggt accttaatta attaaggtac caggtaagtg tacccaattc

     1501 gccctatagt gagtcgtatt acaattcact cgatcgccct tcccaacagt tgcgcagcct

     1561 gaatggcgaa tggagatcca atttttaagt gtataatgtg ttaaactact gattctaatt

     1621 gtttgtgtat tttagattca cagtcccaag gctcatttca ggcccctcag tcctcacagt

     1681 ctgttcatga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac

     1741 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg

     1801 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa

     1861 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc

     1921 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc

     1981 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga

     2041 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc

     2101 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc

     2161 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag

     2221 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa

     2281 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac

     2341 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa

     2401 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat

     2461 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagaatc ctgaggcgga

     2521 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca

     2581 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca

     2641 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc

     2701 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc

     2761 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta

     2821 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga

     2881 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc

     2941 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga

     3001 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct

     3061 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac

     3121 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct

     3181 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt

     3241 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt

     3301 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt

     3361 cgatcaggat gatctggacg aagaacatca ggggctcgcg ccagccgaac tgttcgccag

     3421 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt

     3481 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg

     3541 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagaacttgg

     3601 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg

     3661 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg

     3721 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat

     3781 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg

     3841 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag

     3901 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt

     3961 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc

     4021 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc

     4081 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata

     4141 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg

     4201 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg

     4261 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt

     4321 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg

     4381 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata

     4441 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca

     4501 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag

     4561 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc

     4621 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga

     4681 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg

     4741 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac

     4801 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg

     4861 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg

     4921 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct

     4981 gtggataacc gtattaccgc c

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