质粒名称: | AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE |
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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 |
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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 |
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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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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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