| 质粒名称: | AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 51509 |
| 存储实验室: | Hongkui Zeng |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV-MCS |
| 载体出品公司: | Stratagene |
| 载体抗性: | 氨苄霉素 |
| 完整质粒大小: | 7634 bp |
| 载体修饰: | A fragment containing the phSyn promoter-FLEX-tdTomato-T2A-synaptophysin fused EGFP-WPRE-bGHpA was swapped in to replace the CMV promoter-MCS-hGHpA sequence. Total insert size including the two ITRs is 5037 bp. |
| 载体类型: | AAV |
| 高拷贝/低拷贝: | 未知 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | tdTomato-T2A-SypEGFP |
| 插入大小: | 1428 bp |
| 基因又名: | tdTomato；EGFP；Synaptophysin |
| 启动子 | phSyn1 |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Not1(未破坏) |
| 3' 克隆位点: | Not1(未破坏) |
| 5' 测序引物 | none |
| 3' 测序引物 | none |
|
文献引用方法 |
|
| 材料和方法部分: | AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE was a gift from Hongkui Zeng (爱迪基因 plasmid # 51509) |
| 参考文献部分: | A mesoscale connectome of the mouse brain. Oh SW, Harris JA, Ng L, Winslow B, Cain N, Mihalas S, Wang Q, Lau C, Kuan L, Henry AM, Mortrud MT, Ouellette B, Nguyen TN, Sorensen SA, Slaughterbeck CR, Wakeman W, Li Y, Feng D, Ho A, Nicholas E, Hirokawa KE, Bohn P, Joines KM, Peng H, Hawrylycz MJ, Phillips JW, Hohmann JG, Wohnoutka P, Gerfen CR, Koch C, Bernard A, Dang C, Jones AR, Zeng H. Nature. 2014 Apr 10;508(7495):207-14. doi: 10.1038/nature13186. Epub 2014 Apr 2. 10.1038/nature13186 PubMed 24695228 |
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AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE载体质粒基本信息
AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE质粒图谱载体图谱和AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE载体序列质粒序列多克隆位点信息
-FLEX-tdTomato-T2A-SypEGFP-WPRE质粒图谱.png)
AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE质粒载体简介
AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE质粒序列载体序列
LOCUS AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE 7634 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Can be used to generate AAV virus that will express cytoplasmic
tdTomato and presynaptic (synaptophysin-fused) EGFP in the presence
of Cre in neurons from the synapsin promoter.
ACCESSION .
VERSION .
KEYWORDS AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7634)
AUTHORS Oh SW, Harris JA, Ng L, Winslow B, Cain N, Mihalas S, Wang Q, Lau C,
Kuan L, Henry AM, Mortrud MT, Ouellette B, Nguyen TN, Sorensen SA,
Slaughterbeck CR, Wakeman W, Li Y, Feng D, Ho A, Nicholas E,
Hirokawa KE, Bohn P, Joines KM, Peng H, Hawrylycz MJ, Phillips JW,
Hohmann JG, Wohnoutka P, Gerfen CR, Koch C, Bernard A, Dang C, Jones
AR, Zeng H
TITLE A mesoscale connectome of the mouse brain.
JOURNAL Nature. 2014 Apr 10;508(7495):207-14. doi: 10.1038/nature13186. Epub
2014 Apr 2.
PUBMED 24695228
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE|
FEATURES Location/Qualifiers
source 1..7634
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..141
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 12..141
/label=AAV2 ITR
promoter 177..624
/label=hSyn promoter
/note="human synapsin I promoter; confers neuron-specific
expression (Kugler et al., 2003)"
protein_bind 657..690
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
protein_bind 701..734
/label=lox2272
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GGATACTT). lox2272 sites are compatible with each
other, but incompatible with loxP or loxN sites."
CDS complement(751..1470)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(796..817)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 1145..1164
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1404..1425
/label=EGFP-N
/note="EGFP, reverse primer"
CDS complement(2419..2472)
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
primer_bind complement(3288..3311)
/label=DsRed1-C
/note="DsRed1, forward primer"
primer_bind complement(3486..3505)
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 3755..3773
/label=mCherry-R
/note="mCherry, reverse primer"
protein_bind complement(3937..3970)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
protein_bind complement(3981..4014)
/label=lox2272
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GGATACTT). lox2272 sites are compatible with each
other, but incompatible with loxP or loxN sites."
misc_feature 4038..4626
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4091..4111)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4509..4520)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(4661..4678)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4667..4874
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 4897..5037
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 4897..5026
/label=AAV2 ITR
rep_origin 5112..5567
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5199..5218)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5409..5430
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5584..5603)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5703..5725
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5763..5781)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5849..5953
/gene="bla"
/label=AmpR promoter
CDS 5954..6814
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(6172..6191)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6985..7573
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7474..7493
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
121 actccatcac taggggttcc tgcggccgca cgcgtctgca gagggccctg cgtatgagtg
181 caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc
241 gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg
301 gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca
361 gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg
421 acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc
481 gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac
541 catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg cagcggagga
601 gtcgtgtcgt gcctgagagc gcagacgcgt ggacggtacc cttcaggtcg acgtcgataa
661 cttcgtatag catacattat acgaagttat ccgaatcgca ataacttcgt ataggatact
721 ttatacgaag ttatttggcg cgcctctaga ttacttgtac agctcgtcca tgccgagagt
781 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
841 tttgctcagg gcggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc
901 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt
961 gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac
1021 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
1081 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
1141 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
1201 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca
1261 ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
1321 ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
1381 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
1441 ggtgaacagc tcctcgccct tgctcaccat tcgcgacatc tgattggaga aggaggtggg
1501 cgcaccctgt tggccgtagc cttgctgccc atagtcgccc tgaggcccgt agccaccgcc
1561 accgccgctg gctggctgcc cgtaatcggg ttgataacca ccctgaggcc cgtaggagtc
1621 ctggggccca tagcctccgg ggccctgccc atagcccgca tcgccgtagg catcgccagg
1681 agctggttgc ttttctgggg cgcctggagg tgcgcgcatg aatggggcgg cccagcctgt
1741 ctccttgaac acgaaccata ggttgccaac ccagagcacc aggttcagga agccaaacac
1801 caccgaggtg ttgagtcctg aagtcacagg gtccctcagt tccttgcatg tgtttcctgt
1861 ctggcggcac ataggcatct ccttgataat gttctctggg tcagtggcca tcttcacatc
1921 ggacaggcct ttggcccagg cggatgagct aactagccac atgaaagcga acactgctgt
1981 ggccaggaag tccatcattg gccctttgtt gttctctcgg tacttgttct gcaggaagat
2041 gtaggtggcc agggccccca tggagtagag gaaggcaaac acagccacgg tgacaaagaa
2101 ttcagccgag gaggagtagt caccaactag gaagatcttg gtagtgcccc ctttaacgca
2161 ggagggtgca tcaaagtaca cttggtgcag cctgaatggg tactcaaatt cgacttcgat
2221 gttgagggca ctctccgtct tgttggcaca ctccacgctc agccgaagct ctccggtgta
2281 gctgccgcac gtagcaaagg cgaagatggc aaagacccac tgcagcacct tcacgaagcc
2341 aaggggctcc ttgaccaccc ggaactgacc cccagccacc agctgattca ccacgtccat
2401 tctagatgag cctggggcag ggccgggatt ctcttccacg tctccgcatg tcagaagact
2461 tcctctgccc tcactagtct tgtacagctc gtccatgccg tacagaaaca gatggtgcct
2521 cccctcgctc cgctcatact gctccacaat ggtgtagtct tcgttatggc tagtaatatc
2581 cagctttgtg tcgacgtaat agtagccagg cagctgcacg ggtttcttgg ccatgtagat
2641 ggttttgaat tcgaccagat aatgccctcc atctttcagc ttcagtgcct ggtggatctc
2701 tcccttcagc acgccgtctc tggggtacag cctttctgtt gaagcctccc agcccattgt
2761 cttcttctgc atgactgggc catcaggggg aaaattggtg ccgcgcattt tcaccttata
2821 gatcagagtg ccgtcctgca gagagctgtc ctgggtgaca gtcaccaggc ccccgtcttc
2881 gaagttcatc actctttccc acttaaaccc ctctgggaag ctcagtttct tgtaatctgg
2941 aatgtcagca gggtgtttca cgtaggcttt acttccatac ataaactgag gagacaggat
3001 gtcccatgcg aatggcagag ggccgccctt tgtcactttc agctttgctg tctgggtgcc
3061 ctcatagggc ctcccttccc cctctccttc aatctcaaac tcgtgcccgt tcatgctgcc
3121 ctccatcctg actttaaacc tcataaactc cttaatcacg gccatgttgt tgtcctcgga
3181 ggaggcggtg ccggagctgc cgctgccggt gctgccggta ccatgcccca ggaacaggtg
3241 gtggcggccc tcggagcgct cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt
3301 gatgtccagc ttggtgtcca cgtagtagta gccgggcagt tgcacgggct tcttggccat
3361 gtagatggtc ttgaactcca ccaggtagtg gccgccgtcc ttcagcttca gggcctggtg
3421 gatctcgccc ttcagcacgc cgtcgcgggg gtacaggcgc tcggtggagg cctcccagcc
3481 catggtcttc ttctgcatta cggggccgtc gggggggaag ttggtgccgc gcatcttcac
3541 cttgtagatc agcgtgccgt cctgcaggga ggagtcctgg gtcacggtca ccagaccgcc
3601 gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcttgta
3661 atcggggatg tcggcggggt gcttcacgta cgccttggag ccgtacatga actgggggga
3721 caggatgtcc caggcgaagg gcagggggcc gcccttggtc accttcagct tggcggtctg
3781 ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcat
3841 ggagccctcc atgcgcacct tgaagcgcat gaactctttg atgacctcct cgcccttgct
3901 caccatacta gtggcgcgcc aacctggccg gccttgataa cttcgtataa tgtatgctat
3961 acgaagttat ccgaatcgca ataacttcgt ataaagtatc ctatacgaag ttatcgaaga
4021 tcatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
4081 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
4141 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
4201 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
4261 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
4321 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
4381 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
4441 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
4501 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
4561 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
4621 ccccgcatcg ataccgagcg ctgctcgaga gatcgatctg cctcgactgt gccttctagt
4681 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
4741 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
4801 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
4861 aggcatgctg gggacacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
4921 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
4981 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
5041 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
5101 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
5161 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
5221 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
5281 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
5341 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
5401 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
5461 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
5521 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
5581 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
5641 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
5701 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
5761 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
5821 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
5881 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
5941 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
6001 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
6061 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
6121 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
6181 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
6241 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
6301 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
6361 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
6421 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
6481 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
6541 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
6601 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
6661 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
6721 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
6781 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
6841 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
6901 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
6961 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
7021 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
7081 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
7141 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
7201 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
7261 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
7321 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
7381 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
7441 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
7501 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
7561 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
7621 ttttgctcac atgt
//
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AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE质粒载体应用举例
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