1136- Desmoplakin-GFP人源基因质粒

基本信息
名称：1136- Desmoplakin-GFP人源基因质粒
别称: Plasmid#32227;DSP;DCWHKTADP
启动子：CMV
复制子: pUC
终止子:  SV40 poly(A) signal
质粒大小：13661bp
质粒标签：C-EGFP
原核抗性: Kan
筛选标记：G418
克隆菌株: DH5a
培养条件: 37度

质粒属性
载体宿主：哺乳细胞
载体用途：蛋白表达
基因种属：人
基因类型：ORF
原核抗性：Kan
真核抗性：Neo/G418
荧光蛋白：绿色

质粒简介
   1136- Desmoplakin-GFP质粒是一个哺乳细胞过表达质粒，CMV启动子驱动Desmoplakin人源基因和EGFP绿色荧光蛋白融合表达。该质粒的参考文献是：Desmoplakin assembly dynamics in four dimensions: multiple phases differentially regulated by intermediate filaments and actin. 

质粒图谱


质粒序列
LOCUS       Exported               13661 bp ds-DNA     circular SYN 25-OCT-2017
DEFINITION  synthetic circular DNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13661)
  AUTHORS   .
  TITLE     Direct Submission
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..13661
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    704..982
                     /label=DP insert 5'region
     misc_feature    983..9600
                     /label=Desmoplakin
     CDS             9614..10333
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    10456..10577
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(10584..11039)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        11066..11170
                     /gene="bla"
                     /label=AmpR promoter
     promoter        11172..11529
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11380..11515
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             11564..12358
                     /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
                     SHLAPAEKVSIMADAMRRLHTLDPATC-PFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    12590..12637
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      12966..13554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagctcaa gcttcgaatt cgcggccgcg tcaacagccc
      661 ctgctccttg ggcccctcca tgccatgccg taatctctcc cacccgacca acaccaacac
      721 ccagctccga cgcagctcct ctgcgccctt gccgccctcc gagccacagc tttcctcccg
      781 ctcctgcccc cggcccgtcg ccgtctccgc gctcgcagcg gcctcgggag ggcccaggta
      841 gcgagcagcg acctcgcgag ccttccgcac tcccgcccgg ttccccggcc gtccgcctat
      901 ccttggcccc ctccgctttc tccgcgccgg cccgcctcgc ttatgcctcg gcgctgagcc
      961 gctctcccga ttgcccgccg acatgagctg caacggaggc tcccacccgc ggatcaacac
     1021 tctgggccgc atgatccgcg ccgagtctgg cccggacctg cgctacgagg tgaccagcgg
     1081 cggcgggggc accagcagga tgtactattc tcggcgcggc gtgatcaccg accagaactc
     1141 ggacggctac tgtcaaaccg gcacgatgtc caggcaccag aaccagaaca ccatccagga
     1201 gctgctgcag aactgctccg actgcttgat gcgagcagag ctcatcgtgc agcctgaatt
     1261 gaagtatgga gatggaatac aactgactcg gagtcgagaa ttggatgagt gttttgccca
     1321 ggccaatgac caaatggaaa tcctcgacag cttgatcaga gagatgcggc agatgggcca
     1381 gccctgtgat gcttaccaga aaaggcttct tcagctccaa gagcaaatgc gagcccttta
     1441 taaagccatc agtgtccctc gagtccgcag ggccagctcc aagggtggtg gaggctacac
     1501 ttgtcagagt ggctctggct gggatgagtt caccaaacat gtcaccagtg aatgtttggg
     1561 gtggatgagg cagcaaaggg cggagatgga catggtggcc tggggtgtgg acctggcctc
     1621 agtggagcag cacattaaca gccaccgggg catccacaac tccatcggcg actatcgctg
     1681 gcagctggac aaaatcaaag ccgacctgcg cgagaaatct gcgatctacc agttggagga
     1741 ggagtatgaa aacctgctga aagcgtcctt tgagaggatg gatcacctgc gacagctgca
     1801 gaacatcatt caggccacgt ccagggagat catgtggatc aatgactgcg aggaggagga
     1861 gctgctgtac gactggagcg acaagaacac caacatcgct cagaaacagg aggccttctc
     1921 catacgcatg agtcaactgg aagttaaaga aaaagagctc aataagctga aacaagaaag
     1981 tgaccaactt gtcctcaatc agcatccagc ttcagacaaa attgaggcct atatggacac
     2041 tctgcagacg cagtggagtt ggattcttca gatcaccaag tgcattgatg ttcatctgaa
     2101 agaaaatgct gcctactttc agttttttga agaggcgcag tctactgaag catacctgaa
     2161 ggggctccag gactccatca ggaagaagta cccctgcgac aagaacatgc ccctgcagca
     2221 cctgctggaa cagatcaagg agctggagaa agaacgagag aaaatccttg aatacaagcg
     2281 tcaggtgcag aacttggtaa acaagtctaa gaagattgta cagctgaagc ctcgtaaccc
     2341 agactacaga agcaataaac ccattattct cagagctctc tgtgactaca aacaagatca
     2401 gaaaatcgtg cataaggggg atgagtgtat cctgaaggac aacaacgagc gcagcaagtg
     2461 gtacgtgacg ggcccgggag gcgttgacat gcttgttccc tctgtggggc tgatcatccc
     2521 tcctccgaac ccactggccg tggacctctc ttgcaagatt gagcagtact acgaagccat
     2581 cttggctctg tggaaccagc tctacatcaa catgaagagc ctggtgtcct ggcactactg
     2641 catgattgac atagagaaga tcagggccat gacaatcgcc aagctgaaaa caatgcggca
     2701 ggaagattac atgaagacga tagccgacct tgagttacat taccaagagt tcatcagaaa
     2761 tagccaaggc tcagagatgt ttggagatga tgacaagcgg aaaatacagt ctcagttcac
     2821 cgatgcccag aagcattacc agaccctggt cattcagctc cctggctatc cccagcacca
     2881 gacagtgacc acaactgaaa tcactcatca tggaacctgc caagatgtca accataataa
     2941 agtaattgaa accaacagag aaaatgacaa gcaagaaaca tggatgctga tggagctgca
     3001 gaagattcgc aggcagatag agcactgcga gggcaggatg actctcaaaa acctccctct
     3061 agcagaccag gggtcttctc accacatcac agtgaaaatt aacgagctta agagtgtgca
     3121 gaatgattca caagcaattg ctgaggttct caaccagctt aaagatatgc ttgccaactt
     3181 cagaggttct gaaaagtact gctatttaca gaatgaagta tttggactat ttcagaaact
     3241 ggaaaatatc aatggtgtta cagatggcta cttaaatagc ttatgcacag taagggcact
     3301 gctccaggct attctccaaa cagaagacat gttaaaggtt tatgaagcca ggctcactga
     3361 ggaggaaact gtctgcctgg acctggataa agtggaagct taccgctgtg gactgaagaa
     3421 aataaaaaat gacttgaact tgaagaagtc gttgttggcc actatgaaga cagaactaca
     3481 gaaagcccag cagatccact ctcagacttc acagcagtat ccactttatg atctggactt
     3541 gggcaagttc ggtgaaaaag tcacacagct gacagaccgc tggcaaagga tagataaaca
     3601 gatcgacttt agattatggg acctggagaa acaaatcaag caattgagga attatcgtga
     3661 taactatcag gctttctgca agtggctcta tgatgctaaa cgccgccagg attccttaga
     3721 atccatgaaa tttggagatt ccaacacagt catgcggttt ttgaatgagc agaagaactt
     3781 gcacagtgaa atatctggca aacgagacaa atcagaggaa gtacaaaaaa ttgctgaact
     3841 ttgcgccaat tcaattaagg attatgagct ccagctggcc tcatacacct caggactgga
     3901 aactctgctg aacataccta tcaagaggac catgattcag tccccttctg gggtgattct
     3961 gcaagaggct gcagatgttc atgctcggta cattgaacta cttacaagat ctggagacta
     4021 ttacaggttc ttaagtgaga tgctgaagag tttggaagat ctgaagctga aaaataccaa
     4081 gatcgaagtt ttggaagagg agctcagact ggcccgagat gccaactcgg aaaactgtaa
     4141 taagaacaaa ttcctggatc agaacctgca gaaataccag gcagagtgtt cccagttcaa
     4201 agcgaagctt gcgagcctgg aggagctgaa gagacaggct gagctggatg ggaagtcggc
     4261 taagcaaaat ctagacaagt gctacggcca aataaaagaa ctcaatgaga agatcacccg
     4321 actgacttat gagattgaag atgaaaagag aagaagaaaa tctgtggaag acagatttga
     4381 ccaacagaag aatgactatg accaactgca gaaagcaagg caatgtgaaa aggagaacct
     4441 tggttggcag aaattagagt ctgagaaagc catcaaggag aaggagtacg agattgaaag
     4501 gttgagggtt ctactgcagg aagaaggcac ccggaagaga gaatatgaaa atgagctggc
     4561 aaaggtaaga aaccactata atgaggagat gagtaattta aggaacaagt atgaaacaga
     4621 gattaacatt acgaagacca ccatcaagga gatatccatg caaaaagagg atgattccaa
     4681 aaatcttaga aaccagcttg atagactttc aagggaaaat cgagatctga aggatgaaat
     4741 tgtcaggctc aatgacagca tcttgcaggc cactgagcag cgaaggcgag ctgaagaaaa
     4801 cgcccttcag caaaaggcct gtggctctga gataatgcag aagaagcagc atctggagat
     4861 agaactgaag caggtcatgc agcagcgctc tgaggacaat gcccggcaca agcagtccct
     4921 ggaggaggct gccaagacca ttcaggacaa aaataaggag atcgagagac tcaaagctga
     4981 gtttcaggag gaggccaagc gccgctggga atatgaaaat gaactgagta aggtaagaaa
     5041 caattatgat gaggagatca ttagcttaaa aaatcagttt gagaccgaga tcaacatcac
     5101 caagaccacc atccaccagc tcaccatgca gaaggaagag gataccagtg gctaccgggc
     5161 tcagatagac aatctcaccc gagaaaacag gagcttatct gaagaaataa agaggctgaa
     5221 gaacactcta acccagacca cagagaatct caggagggtg gaagaagaca tccaacagca
     5281 aaaggccact ggctctgagg tgtctcagag gaaacagcag ctggaggttg agctgagaca
     5341 agtcactcag atgcgaacag aggagagcgt aagatataag caatctcttg atgatgctgc
     5401 caaaaccatc caggataaaa acaaggagat agaaaggtta aaacaactga tcgacaaaga
     5461 aacaaatgac cggaaatgcc tggaagatga aaacgcgaga ttacaaaggg tccagtatga
     5521 cctgcagaaa gcaaacagta gtgcgacgga gacaataaac aaactgaagg ttcaggagca
     5581 agaactgaca cgcctgagga tcgactatga aagggtttcc caggagagga ctgtgaagga
     5641 ccaggatatc acgcggttcc agaactctct gaaagagctg cagctgcaga agcagaaggt
     5701 ggaagaggag ctgaatcggc tgaagaggac cgcgtcagaa gactcctgca agaggaagaa
     5761 gctggaggaa gagctggaag gcatgaggag gtcgctgaag gagcaagcca tcaaaatcac
     5821 caacctgacc cagcagctgg agcaggcatc cattgttaag aagaggagtg aggatgacct
     5881 ccggcagcag agggacgtgc tggatggcca cctgagggaa aagcagagga cccaggaaga
     5941 gctgaggagg ctctcttctg aggtcgaggc cctgaggcgg cagttactcc aggaacagga
     6001 aagtgtcaaa caagctcact tgaggaatga gcatttccag aaggcgatag aagataaaag
     6061 cagaagctta aatgaaagca aaatagaaat tgagaggctg cagtctctca cagagaacct
     6121 gaccaaggag cacttgatgt tagaagaaga actgcggaac ctgaggctgg agtacgatga
     6181 cctgaggaga ggacgaagcg aagcggacag tgataaaaat gcaaccatct tggaactaag
     6241 gagccagctg cagatcagca acaaccggac cctggaactg caggggctga ttaatgattt
     6301 acagagagag agggaaaatt tgagacagga aattgagaaa ttccaaaagc aggctttaga
     6361 ggcatctaat aggattcagg aatcaaagaa tcagtgtact caggtggtac aggaaagaga
     6421 gagccttctg gtgaaaatca aagtcctgga gcaagacaag gcaaggctgc agaggctgga
     6481 ggatgagctg aatcgtgcaa aatcaactct agaggcagaa accagggtga aacagcgcct
     6541 ggagtgtgag aaacagcaaa ttcagaatga cctgaatcag tggaagactc aatattcccg
     6601 caaggaggag gctattagga agatagaatc ggaaagagaa aagagtgaga gagagaagaa
     6661 cagtcttagg agtgagatcg aaagactcca agcagagatc aagagaattg aagagaggtg
     6721 caggcgtaag ctggaggatt ctaccaggga gacacagtca cagttagaaa cagaacgctc
     6781 ccgatatcag agggagattg ataaactcag acagcgccca tatgggtccc atcgagagac
     6841 ccagactgag tgtgagtgga ccgttgacac ctccaagctg gtgtttgatg ggctgaggaa
     6901 gaaggtgaca gcaatgcagc tctatgagtg tcagctgatc gacaaaacaa ccttggacaa
     6961 actattgaag gggaagaagt cagtggaaga agttgcttct gaaatccagc cattccttcg
     7021 gggtgcagga tctatcgctg gagcatctgc ttctcctaag gaaaaatact ctttggtaga
     7081 ggccaagaga aagaaattaa tcagcccaga atccacagtc atgcttctgg aggcccaggc
     7141 agctacaggt ggtataattg atccccatcg gaatgagaag ctgactgtcg acagtgccat
     7201 agctcgggac ctcattgact tcgatgaccg tcagcagata tatgcagcag aaaaagctat
     7261 cactggtttt gatgatccat tttcaggcaa gacagtatct gtttcagaag ccatcaagaa
     7321 aaatttgatt gatagagaaa ccggaatgcg cctgctggaa gcccagattg cttcaggggg
     7381 tgtagtagac cctgtgaaca gtgtcttttt gccaaaagat gtcgccttgg cccgggggct
     7441 gattgataga gatttgtatc gatccctgaa tgatccccga gatagtcaga aaaactttgt
     7501 ggatccagtc accaaaaaga aggtcagtta cgtgcagctg aaggaacggt gcagaatcga
     7561 accacatact ggtctgctct tgctttcagt acagaagaga agcatgtcct tccaaggaat
     7621 cagacaacct gtgaccgtca ctgagctagt agattctggt atattgagac cgtccactgt
     7681 caatgaactg gaatctggtc agatttctta tgacgaggtt ggtgagagaa ttaaggactt
     7741 cctccagggt tcaagctgca tagcaggcat atacaatgag accacaaaac agaagcttgg
     7801 catttatgag gccatgaaaa ttggcttagt ccgacctggt actgctctgg agttgctgga
     7861 agcccaagca gctactggct ttatagtgga tcctgttagc aacttgaggt taccagtgga
     7921 ggaagcctac aagagaggtc tggtgggcat tgagttcaaa gagaagctcc tgtctgcaga
     7981 acgagctgtc actgggtata atgatcctga aacaggaaac atcatctctt tgttccaagc
     8041 catgaataag gaactcatcg aaaagggcca cggtattcgc ttattagaag cacagatcgc
     8101 aaccgggggg atcattgacc caaaggagag ccatcgttta ccagttgaca tagcatataa
     8161 gaggggctat ttcaatgagg aactcagtga gattctctca gatccaagtg atgataccaa
     8221 aggatttttt gaccccaaca ctgaagaaaa tcttacctat ctgcaactaa aagaaagatg
     8281 cattaaggat gaggaaacag ggctctgtct tctgcctctg aaagaaaaga agaaacaggt
     8341 gcagacatca caaaagaata ccctcaggaa gcgtagagtg gtcatagttg acccagaaac
     8401 caataaagaa atgtctgttc aggaggccta caagaagggc ctaattgatt atgaaacctt
     8461 caaagaactg tgtgagcagg aatgtgaatg ggaagaaata accatcacgg gatcagatgg
     8521 ctccaccagg gtggtcctgg tagatagaaa gacaggcagt cagtatgata ttcaagatgc
     8581 tattgacaag ggccttgttg acaggaagtt ctttgatcag taccgatccg gcagcctcag
     8641 cctcactcaa tttgctgaca tgatctcctt gaaaaatggt gtcggcacca gcagcagcat
     8701 gggcagtggt gtcagcgatg atgtttttag cagctcccga catgaatcag taagtaagat
     8761 ttccaccata tccagcgtca ggaatttaac cataaggagc agctcttttt cagacaccct
     8821 ggaagaatcg agccccattg cagccatctt tgacacagaa aacctggaga aaatctccat
     8881 tacagaaggt atagagcggg gcatcgttga cagcatcacg ggtcagaggc ttctggaggc
     8941 tcaggcctgc acaggtggca tcatccaccc aaccacgggc cagaagctgt cacttcagga
     9001 cgcagtctcc cagggtgtga ttgaccaaga catggccacc aggctgaagc ctgctcagaa
     9061 agccttcata ggcttcgagg gtgtgaaggg aaagaagaag atgtcagcag cagaggcagt
     9121 gaaagaaaaa tggctcccgt atgaggctgg ccagcgcttc ctggagttcc agtacctcac
     9181 gggaggtctt gttgacccgg aagtgcatgg gaggataagc accgaagaag ccatccggaa
     9241 ggggttcata gatggccgcg ccgcacagag gctgcaagac accagcagct atgccaaaat
     9301 cctgacctgc cccaaaacca aattaaaaat atcctataag gatgccataa atcgctccat
     9361 ggtagaagat atcactgggc tgcgccttct ggaagccgcc tccgtgtcgt ccaagggctt
     9421 acccagccct tacaacatgt cttcggctcc ggggtcccgc tccggctccc gctcgggatc
     9481 tcgctccgga tctcgctccg ggtcccgcag tgggtcccgg agaggaagct ttgacgccac
     9541 agggaattct tcctactctt attcctactc atttagcagt agttctattg ggcacgatcc
     9601 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     9661 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     9721 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     9781 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     9841 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     9901 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     9961 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
    10021 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
    10081 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
    10141 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
    10201 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
    10261 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
    10321 gctgtacaag taaagcggcc gcgactctag atcataatca gccataccac atttgtagag
    10381 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
    10441 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
    10501 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
    10561 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
    10621 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
    10681 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
    10741 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
    10801 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
    10861 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
    10921 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
    10981 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
    11041 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
    11101 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
    11161 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
    11221 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
    11281 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
    11341 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
    11401 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
    11461 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
    11521 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
    11581 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
    11641 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
    11701 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
    11761 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
    11821 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
    11881 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
    11941 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
    12001 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
    12061 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
    12121 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
    12181 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
    12241 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
    12301 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
    12361 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
    12421 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
    12481 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
    12541 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
    12601 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
    12661 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
    12721 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
    12781 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
    12841 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
    12901 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
    12961 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
    13021 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
    13081 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
    13141 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
    13201 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
    13261 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
    13321 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
    13381 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
    13441 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
    13501 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
    13561 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
    13621 cctgcgttat cccctgattc tgtggataac cgtattaccg c
//

质粒菌株产品操作说明书
一、扩增流程
         收到产品后，请先根据产品管壁标签来判断产品形式，并在扩增前准确查找该质粒菌株的抗性、感受态和培养温度。
         1、质粒干粉（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
              ①收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O去离子水溶解质粒；
              ②取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；（从第二步开始均要在超净工作台中无菌操作）
              ③加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min；
              ④6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上；（可使用本平台的平板涂布专用玻璃珠进行涂布，可以比传统涂布方法获得更多转化子）
              ⑤将平板正向培养1h，再倒置37℃培养14h。如果要求是30度则培养20h；
（菌落过多则将质粒稀释后再转化。没有菌落则加入10μl质粒转化。另不要直接转表达感受态，要先转克隆感受态，重提质粒后再导入表达感受态）
              ⑥挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
        2、甘油菌种（冰袋运输，存于-80℃，保质期90天，请务必划线挑单克隆培养）
              四区划线后挑单菌落培养，酵母菌需要先液体复苏再四区划线，再挑单菌落液体培养。
        3、穿刺菌种（冰袋运输，存于4℃，保质期7天）
              穿刺接种，液体培养后四区划线，再挑单菌落液体培养。
        4、菌落平板（冰袋运输，存于4℃，保质期7天）
              直接挑取单菌落至液体培养基中。
        5、液体质粒（冰袋运输，存于-20℃，保质期90天）
              单独提取的液体质粒收到后可直接使用。
        6、滤纸质粒（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
              收到货后将滤纸画圈部分剪下放入EP管中，加100ul无菌水将滤纸浸湿并浸泡5min，吸取5ul质粒转化，离心全涂。

二、转化图片

P1169/pECMV-3×FLAG-H-FABP人源基因质粒

P1170/pECMV-3×FLAG-TRAF6-S12F人源基因质粒

P1171/pECMV-3×FLAG-GSK3B人源基因质粒

P1172/pECMV-3×FLAG-CD1D人源基因质粒

P1719/pECMV-3×FLAG-CD4人源基因质粒

P1830/pECMV-3×FLAG-CD22人源基因质粒

P1195/pECMV-3×FLAG-AFP人源基因质粒

P4834/pCDNA3.1-AFP人源基因质粒

P4100/pECMV-3×FLAG-CCNE2人源基因质粒

P4270/pECMV-3×FLAG-AATF人源基因质粒

P4268/pECMV-3×FLAG-BLVRA(1同义突变）人源基因质粒