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載體描述

在pIRES2-DsRed-EXPRESS的基礎上改造而成，2A信號肽序列替換pIRES2-DsRed-Express上的IRES序列。

載體序列

LOCUS       Exported                4769 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 4769)

  AUTHORS   BT-Lab

  TITLE     Direct Submission

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

FEATURES             Location/Qualifiers

     source          1..4769

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     enhancer        61..364

                     /label=CMV enhancer

                     /note="human cytomegalovirus immediate early enhancer"

     promoter        365..568

                     /label=CMV promoter

                     /note="human cytomegalovirus (CMV) immediate early

                     promoter"

     misc_feature    591..665

                     /label=MCS

                     /note="multiple cloning site"

     CDS             684..749

                     /codon_start=1

                     /product="2A signal peptide from foot-and-mouth disease

                     virus polyprotein"

                     /label=F2A

                     /note="Eukaryotic ribosomes fail to insert a peptide bond

                     between the Gly and Pro residues, yielding separate

                     polypeptides."

                     /translation="VKQTLNFDLLKLAGDVESNPGP"

     CDS             759..1436

                     /codon_start=1

                     /product="rapidly maturing tetrameric variant of DsRed

                     fluorescent protein (Bevis and Glick, 2002)"

                     /label=DsRed-Express

                     /note="mammalian codon-optimized"

                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK

                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV

                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK

                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH

                     LFL"

     polyA_signal    1557..1678

                     /label=SV40 poly(A) signal

                     /note="SV40 polyadenylation signal"

     rep_origin      complement(1685..2140)

                     /direction=LEFT

                     /label=f1 ori

                     /note="f1 bacteriophage origin of replication; arrow

                     indicates direction of (+) strand synthesis"

     promoter        2167..2271

                     /gene="bla"

                     /label=AmpR promoter

     promoter        2273..2630

                     /label=SV40 promoter

                     /note="SV40 enhancer and early promoter"

     rep_origin      2481..2616

                     /label=SV40 ori

                     /note="SV40 origin of replication"

     CDS             2665..3459

                     /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    3691..3738

                     /label=HSV TK poly(A) signal

                     /note="herpes simplex virus thymidine kinase

                     polyadenylation signal (Cole and Stacy, 1985)"

     rep_origin      4067..4655

                     /direction=RIGHT

                     /label=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 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg

      661 gatccatgaa aattgtcgct cctgtcaaac aaactcttaa ctttgattta ctcaaactgg

      721 ctggggatgt agaaagcaat ccaggtccag ccacaaccat ggcctcctcc gaggacgtca

      781 tcaaggagtt catgcgcttc aaggtgcgca tggagggctc cgtgaacggc cacgagttcg

      841 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg

      901 tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag ttccagtacg

      961 gctccaaggt gtacgtgaag caccccgccg acatccccga ctacaagaag ctgtccttcc

     1021 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga

     1081 cccaggactc ctccctgcag gacggctcct tcatctacaa ggtgaagttc atcggcgtga

     1141 acttcccctc cgacggcccc gtaatgcaga agaagactat gggctgggag gcctccaccg

     1201 agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccacaaggcc ctgaagctga

     1261 aggacggcgg ccactacctg gtggagttca agtctatcta tatggccaag aagcccgtgc

     1321 agctgcccgg ctactactac gtggactcca agctggacat cacctcccac aacgaggact

     1381 acaccatcgt ggagcagtac gagcgcgccg agggccgcca ccacctgttc ctgtagcggc

     1441 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa

     1501 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact

     1561 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata

     1621 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag

     1681 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc

     1741 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc

     1801 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac

     1861 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca

     1921 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg

     1981 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag

     2041 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc

     2101 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga

     2161 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc

     2221 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg

     2281 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag

     2341 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc

     2401 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag

     2461 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc

     2521 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc

     2581 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa

     2641 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg

     2701 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct

     2761 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac

     2821 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg

     2881 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg

     2941 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa

     3001 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca

     3061 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt

     3121 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc

     3181 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc

     3241 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg

     3301 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt

     3361 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag

     3421 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa

     3481 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct

     3541 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg

     3601 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg

     3661 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg

     3721 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata

     3781 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc

     3841 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca

     3901 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa

     3961 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt

     4021 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt

     4081 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt

     4141 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga

     4201 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag

     4261 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata

     4321 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg

     4381 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga

     4441 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca

     4501 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa

     4561 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt

     4621 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac

     4681 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt

     4741 ctgtggataa ccgtattacc gccatgcat

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