pDsRed-Monomer-N1哺乳荧光质粒

基本信息

启动子:	CMV
复制子:	pUC ori,f1 ori
终止子:	SV40 poly(A) signal
质粒分类:	哺乳细胞，荧光蛋白报告载体
质粒大小:	4691bp
原核抗性:	Kan
筛选标记:	Neo
克隆菌株:	DH5α
培养条件:	37℃，有氧 LB
表达宿主:	哺乳细胞
诱导方式:	无须诱导，瞬时表达
5'测序引物:	CMV-F：CGCAAATGGGCGGTAGGCGTG
3'测序引物:	根据序列设计引物

质粒简介

      pDsRed-Monomer-N1 is a mammalian expression vector that encodes DsRed-Monomer (DsRed.M1), a monomeric mutant derived from the tetrameric Discosoma sp. red fuorescent protein DsRed. DsRed-Monomer contains forty-fve amino acid substitutions (listed on page 2). When DsRed-Monomer is expressed in mammalian cell cultures, red fuorescent cells can be detected by either fuorescence microscopy or fow cytometry 12–16 hr after transfection (DsRed-Monomer excitation and emission maxima = 557 nm and 592 nm, respectively). The DsRed-Monomer coding sequence is human codon-optimized for high expression in mammalian cells.

质粒图谱

质粒序列

LOCUS       Exported                4691 bp ds-DNA    circular SYN 22-10-2015
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4691)
  AUTHORS   admin
  TITLE     Direct Submission
  JOURNAL   Exported 2015-10-22  from MLCC
FEATURES             Location/Qualifiers
     source          1..4691
                     /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"
     misc_feature    591..671
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             679..1356
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Strongin et al., 2007)"
                     /note="DsRed-Monomer"
                     /note="mammalian codon-optimized"
                     /translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGV
                     VEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISH
                     ALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHS
                     GSQ"
     polyA_signal    1479..1600
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1607..2062)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2089..2193
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        2195..2552
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      2403..2538
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             2587..3381
                     /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    3613..3660
                     /note="HSV TK poly(A) signal"
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     rep_origin      3989..4577
                     /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 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      661 gatccaccgg tcgccaccat ggacaacacc gaggacgtca tcaaggagtt catgcagttc
      721 aaggtgcgca tggagggctc cgtgaacggc cactacttcg agatcgaggg cgagggcgag
      781 ggcaagccct acgagggcac ccagaccgcc aagctgcagg tgaccaaggg cggccccctg
      841 cccttcgcct gggacatcct gtccccccag ttccagtacg gctccaaggc ctacgtgaag
      901 caccccgccg acatccccga ctacatgaag ctgtccttcc ccgagggctt cacctgggag
      961 cgctccatga acttcgagga cggcggcgtg gtggaggtgc agcaggactc ctccctgcag
     1021 gacggcacct tcatctacaa ggtgaagttc aagggcgtga acttccccgc cgacggcccc
     1081 gtaatgcaga agaagactgc cggctgggag ccctccaccg agaagctgta cccccaggac
     1141 ggcgtgctga agggcgagat ctcccacgcc ctgaagctga aggacggcgg ccactacacc
     1201 tgcgacttca agaccgtgta caaggccaag aagcccgtgc agctgcccgg caaccactac
     1261 gtggactcca agctggacat caccaaccac aacgaggact acaccgtggt ggagcagtac
     1321 gagcacgccg aggcccgcca ctccggctcc cagtagagcg gccgcgactc tagatcataa
     1381 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     1441 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     1501 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1561 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     1621 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     1681 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     1741 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     1801 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     1861 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     1921 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     1981 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2041 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2101 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2161 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2221 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2281 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2341 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2401 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2461 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2521 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     2581 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2641 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     2701 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     2761 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     2821 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     2881 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     2941 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3001 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3061 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3121 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3181 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3241 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3301 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3361 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3421 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3481 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3541 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3601 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     3661 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     3721 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     3781 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     3841 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3901 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3961 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4021 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4081 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4141 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4201 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4261 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4321 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4381 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4441 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4501 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4561 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4621 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4681 ccgccatgca t