sfGFP-N1

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sfGFPN1载体基本信息

质粒名称: sfGFP-N1 
出品公司: 爱迪基因
目录编号: 54737
存储实验室: Michael Davidson, Geoffrey Waldo

载体骨架基本信息
载体名称: sfGFP-N1
载体出品公司: --
载体抗性: 卡那霉素
空载体大小: 4750 bp
完整质粒大小: --
载体修饰: --
载体类型: 哺乳细胞表达
筛选标记: 新霉素G418
高拷贝/低拷贝: 高拷贝
生长条件: 37 ℃

插入基因信息
插入基因名称: --
物种来源: --
插入大小: --
基因突变: --
GenBank编号 --
启动子 --
标签/融合蛋白: sfGFP(主干上的C终端)
克隆方法: --
5' 克隆位点: --
3' 克隆位点: --
5' 测序引物 --
3' 测序引物 --

文献引用方法
材料和方法部分: sfGFP-N1 was a gift from Michael Davidson & Geoffrey Waldo (Addgene plasmid # 54737 ; http://n2t.net/addgene:54737 ; RRID:Addgene_54737)
参考文献部分: Engineering and characterization of a superfolder green fluorescent protein. Pédelacq JD, Cabantous S, Tran T, Terwilliger TC, Waldo GS. Nat Biotechnol. 2006 Jan . 24(1):79-88. 10.1038/nbt1172 PubMed 16369541

sfGFPN1载体图谱质粒图谱和多克隆位点信息

sfGFP-N1质粒图谱

sfGFPN1载体简介

sfGFPN1载体序列

LOCUS       sfGFP-N1                4732 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Localization: N1 Cloning Vector, Excitation: 485, Emission: 507.
ACCESSION   .
VERSION     .
KEYWORDS    sfGFP-N1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4732)
  TITLE     Michael Davidson Empty Backbones
REFERENCE   2  (bases 1 to 4732)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4732
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    531..611
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      613..622
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             619..1338
                     /codon_start=1
                     /product="enhanced CFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             619..1338
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(664..685)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1272..1293
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1460..1581
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1497..1516)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1551..1570
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1588..2043)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1725..1746)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1937..1956
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2070..2174
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2176..2533
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2199..2219)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2384..2519
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2446..2465
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2568..3362
                     /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"
     primer_bind     complement(2622..2641)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3232..3251
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3550..3569)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3594..3641
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3970..4558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4459..4478
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      601 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      661 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgcg cggcgagggc
      721 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      781 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
      841 taccccgacc acatgaagcg ccacgacttc ttcaagtccg ccatgcccga aggctacgtc
      901 caggagcgca ccatcagctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag
      961 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1021 ggcaacatcc tggggcacaa gctggagtac aacttcaaca gccacaacgt ctatatcacc
     1081 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa cgtggaggac
     1141 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1201 ctgctgcccg acaaccacta cctgagcacc cagtccgtgc tgagcaaaga ccccaacgag
     1261 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tcacggcatg
     1321 gacgagctgt acaagtaagc ggccgcgact ctagatcata atcagccata ccacatttgt
     1381 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     1441 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     1501 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     1561 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     1621 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     1681 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     1741 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     1801 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     1861 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     1921 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     1981 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     2041 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2101 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2161 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     2221 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2281 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2341 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2401 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2461 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2521 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     2581 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     2641 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     2701 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     2761 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     2821 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     2881 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     2941 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3001 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     3061 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     3121 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     3181 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     3241 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     3301 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     3361 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     3421 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     3481 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     3541 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     3601 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     3661 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     3721 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     3781 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     3841 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3901 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3961 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4021 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4081 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4141 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4201 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4261 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4321 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4381 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4441 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4501 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4561 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4621 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     4681 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cg
//

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