| 质粒名称: | pAAV-Ef1a-DIO ChETA-EYFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 26968 |
| 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: | 氨苄霉素 |
| 空载体大小: | 5587 bp |
| 完整质粒大小: | 7249 bp |
| 载体类型: | 哺乳细胞表达，AAV |
| 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: | hChR2(E123T/H134R)-EYFP |
| 物种来源: | 人源 |
| 插入大小: | 1662 bp |
| 基因又名: | ChETA-EYFP |
| 基因突变: | E123T, H134R |
| 标签/融合蛋白: | EYFP (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AscI(未破坏) |
| 3' 克隆位点: | NheI(未破坏) |
| 5' 测序引物 | ggccagcttggcacttgatg |
| 3' 测序引物 | GCAATAGCATGATACAAAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-Ef1a-DIO ChETA-EYFP was a gift from Karl Deisseroth (爱迪基因 plasmid # 26968) |
| 参考文献部分: | Ultrafast optogenetic control. Gunaydin LA, Yizhar O, Berndt A, Sohal VS, Deisseroth K, Hegemann P. Nat Neurosci. 2010 Mar . 13(3):387-92. 10.1038/nn.2495 PubMed 20081849 |
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-Ef1a-DIO ChETA-EYFP 7256 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION AAV expression of humanized ChR2 with E123T/H134R mutations fused to
EYFP driven by EF1a promoter for optogenetic activation.
ACCESSION .
VERSION .
KEYWORDS pAAV-Ef1a-DIO ChETA-EYFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7256)
AUTHORS Gunaydin LA, Yizhar O, Berndt A, Sohal VS, Deisseroth K, Hegemann P
TITLE Ultrafast optogenetic control.
JOURNAL Nat Neurosci. 2010 Mar . 13(3):387-92.
PUBMED 20081849
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pAAV-Ef1a-DIO ChETA-EYFP|
FEATURES Location/Qualifiers
source 1..7256
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 51..180
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
promoter 281..1459
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 512..1450
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1407..1427
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
protein_bind 1490..1523
/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."
protein_bind 1574..1607
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS complement(1619..2338)
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1664..1685)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 2013..2032
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2272..2293
/label=EGFP-N
/note="EGFP, reverse primer"
regulatory 2335..2344
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 3277..3286
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
protein_bind complement(3291..3324)
/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."
protein_bind complement(3375..3408)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 3433..4021
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3486..3506)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3904..3915)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 4053..4529
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(4252..4271)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 4569..4709
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 4569..4698
/label=AAV2 ITR
rep_origin 4784..5239
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4871..4890)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5081..5102
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5256..5275)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5375..5397
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5435..5453)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5521..5625
/gene="bla"
/label=AmpR promoter
CDS 5626..6486
/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(5844..5863)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6657..7245
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7146..7165
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
61 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
121 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
181 gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga ggaatctttg
241 cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg
301 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
361 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
421 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
481 ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc
541 tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta
601 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
661 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
721 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
781 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
841 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
901 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
961 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
1021 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
1081 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
1141 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
1201 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
1261 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
1321 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
1381 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
1441 tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa taacttcgta
1501 taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc tattagcaat
1561 atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc gcgccctatt
1621 acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
1681 ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactggtag ctcaggtagt
1741 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
1801 cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt
1861 tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
1921 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
1981 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
2041 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
2101 tcatgtggtc ggggtagcgg gcgaagcact gcaggccgta gccgaaggtg gtcacgaggg
2161 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
2221 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
2281 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
2341 tggcggccgc tggcacggct ccggcctcgg cttcgtcttc gacgagagtc tcgacctcga
2401 tctccgttcc gccgatgttc agtttggtgg ttttgcggat atctccgtga atcaatatgt
2461 gctcgtggat caggacgcgc aggtagtgtc ccaacaaccc ccaacaattt ttactcatca
2521 gatcaataat cgtgtgacct acggtggagc catagacgct caggacgcca aaaccttcgg
2581 gccccaaaat gaagagaatt gggaacatac cccagctcac gaaaaacagc catgccatgc
2641 cggtcacgac ctggcggcac cgaccctttg gcacagtatg ataaccctcg atatatgctt
2701 tggcggcgtg aaaaaatgtg ttcgcgccat agcacaatcc aagacaaaag aagatgactt
2761 taacatagcc ggttgccatg gcgctggtag ccccccacac gatagtcccg atgtctgaga
2821 caaggagtcc catggttctc ctgctgtagt cgttgctcag gccggtgagg ttgctcaggc
2881 ggataaggat gacaggacaa gtgagcagcc aggttgcata gcgcagccac tgcacccggt
2941 gtcctgtggc aaggtagagc atagagggat tcttaaactc aaaaaagaac tcgagaatca
3001 ccttaaccat ttcaatggcg cacacataga tctcctccca gccgcatgta gatttccagg
3061 tttggtaggc atagaacatc agcagcaaaa tgctgaatcc tgctgcaagc cactgcagga
3121 catttgacgc ggtctgagcg ccgttcgtgc cgcgagattc aatccatccg gcacagtaac
3181 attgatcctc agggaccagg acggacccgt tcaccaccac aggattagta acgaacaaaa
3241 gttcgcgtcc gacggcagac aaagcgccgc catagtccat ggtggctagc ataacttcgt
3301 ataaagtatc ctatacgaag ttatttgcct taacccagaa attatcactg ttattcttta
3361 gaatggtgca aagaataact tcgtataatg tatgctatac gaagttatga attcgatatc
3421 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
3481 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
3541 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
3601 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
3661 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
3721 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
3781 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
3841 ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
3901 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
3961 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
4021 catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag
4081 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
4141 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
4201 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
4261 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
4321 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
4381 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
4441 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
4501 cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga
4561 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
4621 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
4681 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
4741 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
4801 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
4861 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
4921 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
4981 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
5041 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
5101 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
5161 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
5221 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
5281 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
5341 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
5401 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
5461 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
5521 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
5581 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
5641 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
5701 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
5761 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
5821 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
5881 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
5941 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
6001 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
6061 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
6121 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
6181 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
6241 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
6301 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
6361 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
6421 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
6481 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
6541 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
6601 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
6661 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
6721 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
6781 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
6841 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
6901 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
6961 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
7021 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
7081 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
7141 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
7201 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaa
//
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