| 质粒名称: | pAAV-hSyn-eNpHR 3.0-EYFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 26972 |
| 存储实验室: | Karl Deisseroth |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4570 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,AAV |
| 筛选标记: | -- |
| 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: | eNpHR 3.0 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1683 bp |
| 基因突变: | Trafficking Signal (TS) ER Export Signal |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | EYFP（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI, AgeI(未破坏) |
| 3' 克隆位点: | EcoRI, HinDIII (未破坏) |
| 5' 测序引物 | ccacgcgaggcgcgagatag |
| 3' 测序引物 | GCAATAGCATGATACAAAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-hSyn-eNpHR 3.0-EYFP was a gift from Karl Deisseroth (Addgene plasmid # 26972 ; http://n2t.net/addgene:26972 ; RRID:Addgene_26972) |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pAAV-hSyn-eNpHR 3.0-EYFP 6242 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION AAV expression of eNpHR 3.0 fused to EYFP driven by humanized
Synapsin promoter for optogenetic inhibition.
ACCESSION .
VERSION .
KEYWORDS pAAV-hSyn-eNpHR 3.0-EYFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6242)
TITLE eNpHR3.0-EYFP
REFERENCE 2 (bases 1 to 6242)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6242
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 52..181
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
promoter 226..673
/label=hSyn promoter
/note="human synapsin I promoter; confers neuron-specific
expression (Kugler et al., 2003)"
CDS 1655..2368
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(1697..1718)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1958..1977)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2305..2326
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 2420..3008
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2473..2493)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2891..2902)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3040..3516
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3239..3258)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 3556..3696
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3556..3685
/label=AAV2 ITR
rep_origin 3771..4226
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3858..3877)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4068..4089
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4243..4262)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4362..4384
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4422..4440)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4508..4612
/gene="bla"
/label=AmpR promoter
CDS 4613..5473
/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(4831..4850)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5644..6232
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6133..6152
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc
61 agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg
121 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
181 tgcggccgca cgcgtgtgtc tagactgcag agggccctgc gtatgagtgc aagtgggttt
241 taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga
301 caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa
361 caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc
421 cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc
481 cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc
541 agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg
601 cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg
661 cctgagagcg cagtcgagaa ggtaccggat ccccgggtac cggtaggcca ccatgacaga
721 gaccctgcct cccgtgaccg agagtgccgt ggcccttcaa gccgaggtta cccaaaggga
781 gttgttcgag ttcgtgctga acgacccttt gcttgcaagc agtctctata tcaacatcgc
841 acttgcagga ctgagtatac tgctgttcgt ttttatgacc cgaggactcg atgatccacg
901 ggcaaaactt attgctgtgt caaccatcct tgtgcctgtc gtcagcattg cctcctacac
961 tggattggcg agcggcctga caatttccgt tcttgaaatg ccagcgggcc attttgcaga
1021 aggcagctca gtgatgctgg gaggagaaga ggtagatggt gtagtcacca tgtggggacg
1081 gtatctcacc tgggcacttt ccacgcccat gattctcctc gctctgggtc tcctggccgg
1141 aagcaatgct acaaagctct tcacagctat cactttcgat atcgctatgt gcgtgactgg
1201 ccttgccgcg gccctgacta cctcctccca cctcatgaga tggttctggt acgctatcag
1261 ttgtgcatgc tttctggtgg tcttgtatat cctgctggtg gagtgggcac aggacgccaa
1321 agccgcggga accgctgaca tgttcaatac cctgaagctg ttgacagtag tgatgtggct
1381 ggggtatcca attgtgtggg ctcttggagt cgagggtatc gcggtgttgc ccgttggggt
1441 gacgagctgg ggatattctt tcctggatat cgtggcaaag tacattttcg cattcttgct
1501 cctgaactat ctgacgtcaa acgaatctgt cgtgtccggc agcattttgg atgttccatc
1561 tgcttctggg accccggctg atgatgcggc cgccaagagc aggatcacca gcgagggcga
1621 gtacatcccc ctggaccaga tcgacatcaa cgtggtgagc aagggcgagg agctgttcac
1681 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
1741 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
1801 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca
1861 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
1921 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
1981 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
2041 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
2101 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
2161 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
2221 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa
2281 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
2341 cactctcggc atggacgagc tgtacaagtt ctgctacgag aacgaggtgt aatgagaatt
2401 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
2461 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
2521 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
2581 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
2641 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
2701 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
2761 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
2821 gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg
2881 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
2941 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
3001 ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc
3061 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
3121 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
3181 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
3241 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
3301 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
3361 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
3421 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
3481 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc
3541 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
3601 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
3661 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
3721 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
3781 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
3841 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
3901 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
3961 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
4021 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
4081 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
4141 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
4201 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
4261 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
4321 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
4381 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
4441 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
4501 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
4561 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
4621 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
4681 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
4741 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
4801 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
4861 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
4921 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
4981 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
5041 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
5101 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
5161 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
5221 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
5281 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
5341 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
5401 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
5461 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
5521 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
5581 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
5641 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
5701 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
5761 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
5821 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
5881 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
5941 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
6001 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
6061 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
6121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
6181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
6241 ca
//
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