| 质粒名称: | AAV-P(Cry1)-DIO-intron336-Venus-NLS-D2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 110055 |
| 存储实验室: | Eric Erquan Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | aav2 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4171 bp |
| 完整质粒大小: | 7683 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，AAV，Cre/Lox |
| 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: | Cry1 promoter and Venus,Cry1 intron336 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | Cry1 |
| 标签/融合蛋白: | NLS-D2 (C terminal on insert) |
| 克隆方法: | Gibson 克隆 |
| 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: | AAV-P(Cry1)-DIO-intron336-Venus-NLS-D2 was a gift from Eric Erquan Zhang (Addgene plasmid # 110055 ; http://n2t.net/addgene:110055 ; RRID:Addgene_110055) |
| 参考文献部分: | Long-term in vivo recording of circadian rhythms in brains of freely moving mice. Mei L, Fan Y, Lv X, Welsh DK, Zhan C, Zhang EE. Proc Natl Acad Sci U S A. 2018 Apr 2. pii: 1717735115. doi: 10.1073/pnas.1717735115. 10.1073/pnas.1717735115 PubMed 29610316 |
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
-DIO-intron336-Venus-NLS-D2质粒图谱.png)
载体简介
载体序列
LOCUS AAV-P(Cry1)-DIO-intron336-Venus-NLS-D2 7672 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Cry1 transcription fluorescence reporter.
ACCESSION .
VERSION .
KEYWORDS AAV-P(Cry1)-DIO-intron336-Venus-NLS-D2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7672)
AUTHORS Mei L, Fan Y, Lv X, Welsh DK, Zhan C, Zhang EE
TITLE Long-term in vivo recording of circadian rhythms in brains of freely
moving mice.
JOURNAL Proc Natl Acad Sci U S A. 2018 Apr 2. pii: 1717735115. doi:
10.1073/pnas.1717735115.
PUBMED 29610316
REFERENCE 2 (bases 1 to 7672)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7672
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..130
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
protein_bind 1722..1755
/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 1806..1839
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS complement(1989..2009)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(2013..2726)
/codon_start=1
/product="Venus YFP with monomerizing A206K mutation (Nagai
et al., 2002)"
/label=mVenus
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(2055..2076)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 2404..2423
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2663..2684
/label=EGFP-N
/note="EGFP, reverse primer"
CDS complement(2736..2756)
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS complement(2781..2825)
/codon_start=1
/product="peptide tag that allows for enzymatic
biotinylation"
/label=AviTag(TM)
/translation="GLNDIFEAQKIEWHE"
CDS complement(2847..2912)
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS complement(3303..3326)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
regulatory 3326..3335
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
promoter complement(3373..3391)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(3374..3391)
/label=SP6
/note="SP6 promoter, forward primer"
promoter complement(3397..3415)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3458..3477)
/label=pMT2-F
/note="Synthetic intron, forward primer"
intron 3460..3592
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
protein_bind complement(3657..3690)
/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(3741..3774)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 3799..4387
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3852..3872)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4270..4281)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 4419..4895
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(4618..4637)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 4935..5075
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 4935..5064
/label=AAV2 ITR
rep_origin 5150..5605
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5237..5256)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5447..5468
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5622..5641)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5741..5763
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5801..5819)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5887..5991
/gene="bla"
/label=AmpR promoter
CDS 5992..6852
/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(6210..6229)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7023..7611
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7512..7531
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
121 aggggttcct gcggccgcac gcgtgtaaag atgcacatgt ggccctgtgt tctcattaga
181 cttcaattgc tttgataaaa catcatgagc aaaagcaact tggccaggaa ggaaggggtt
241 tgttgccgtt tacagttaat aatccatcgt ggggagaagt cagggcagaa attcaaagca
301 gaaacctgga gatagagggc tgaagcagaa gccatggagg aaagccgttt tggttccctg
361 tggcttcctc agcctgcttc catatacaac ccgcctaggg ctaccagcct aatggtggcg
421 ccacacacac tggtcgggtc cttccacatc actcattaat caagaaagtg cccagacttc
481 cctacaggca acctgatgga agtctttact acaattaagg ttcctcttcc catttaactt
541 cagttgaaga aaaaaaaaaa aaaaaaaaaa aaaagcaaag caaaaagccc aaaaacctaa
601 tcaaaatacc ttctaaacta cacttcatgt aatagattag agttgataaa ttgacgatgt
661 atctacagta ttatgaaatg ctgaatatct gagatcggac ctgtcttctc cagagcttgg
721 gcaaaggtgc ccgccccaaa cagtaaaaaa ggctaacggc gtgatttctg ctcctttcag
781 agcaggggac tccgattaac acctgattga agctgggtga gaggcaggac tgtgcctcgg
841 ttgcctctca cccagcccag caataactct cagggcacac ggaactggct tcagaccttc
901 cacacaaccc ataggaagag ccctgggcgc gctcccgcgt cctagcctct tctcttccac
961 cgcgagagcg cagcgcacgc gcgccgtgcc cggcagagcg cgcgcccgcg agctccccgc
1021 agccccgtgc tcatccgggt ccccgcgagc ctcggccaat ggaggcggct cccggccccg
1081 ggagcagaac tatgcctcct cccccttctc gcggcctgtg gcccccgcga gcggcctggg
1141 gctcaggaga cccgcagagg ctgcgagtcc ggatcccacg cgagaactca ggtcgtgagc
1201 gcccagcgat cgcaaccggt cccgaagcac cgggcgcggc ggcgcacgcg ggggtctgag
1261 ccagtgtagt aaacacactt cagaaacgtg aggtgccggt ggtcacgtgg ggtgcatgcc
1321 gcccaatgag aagccggggg cggggcggag gccgctgacg cggcggagtc acccgggcag
1381 cctcgggacc ggtgaccggc aggcgagcgt cccgcggcct ggagctctag cccgcgctct
1441 ccgagctcgc tgcccactcc gggacgcaca ggccgcgtcg cgcggaggcc gcgtgagctc
1501 gtgtccgttc gtggaggccg cctcgggcgc gccaccgtct tctcgcgggt cgcctcggtt
1561 gggcgaggtt tccgtgacga gcctcccggg gctgctgcga ccagcccggg ccgtcgtggc
1621 ggctcgggcc ccgcgcgctc gcttgcgctc cgggggtctc aggccctgga aggtatgcgt
1681 gtcctccgga cgagatctgc gatctaagtg tcgactccgg aataacttcg tataggatac
1741 tttatacgaa gttatgcaga atggtagctg gattgtagct gctattagca atatgaaacc
1801 tcttaataac ttcgtatagc atacattata cgaagttatg gctacacatt gatcctagca
1861 gaagcacagg ctgcagggtg acggtccatc ccgctctcct gggcacaaga catgggcagc
1921 gtgccatcat cctgctcctc cacctccggc gggaagccat ggctaagctt agaattcacg
1981 gggtcttcta cctttctctt cttttttgga ggcttgtaca gctcgtccat gccgagagtg
2041 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
2101 ttgctcagtt tggactggta gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
2161 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg
2221 tggcggatct tgaagtttgc cttgatgccg ttcttctgct tgtcggcggt gatatagacg
2281 ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
2341 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
2401 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
2461 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac
2521 tgcaggccgt agccgagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
2581 gtgcagatca acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
2641 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
2701 gtgaacagct cctcgccctt gctcacacct gcagggccct gaaaatacag gttttcggat
2761 cctgaaccgc ctgccgatcc ctcatgccac tctattttct gagcttcgaa aatatcgttc
2821 aggccgctag ctgcggccgc tccaccctta tcgtcatcgt ccttgtagtc gatgtcatga
2881 tctttataat caccgtcatg gtctttgtag tccatggtgg gtacctcttg acagctctac
2941 cttttactac tataaaaacg tacatgtagt aacttcctta tgccacttcc aaaagatgat
3001 ttcaacatga atcaacagaa atagtttcta aagtaggtca tcgctatagc aaccagtggg
3061 tggtttagca tactcccatc acaatgacct actttctagt ctgtgctagg acagcttata
3121 taactagtat ttctagttct gtctataaaa tagtctgaaa aaggcaatgg caagatacaa
3181 cataatccca ctgctactga aaacaagaca ttatgaattc atataacaca gtcatctgtc
3241 ctatgccata gtttctacac tgtgtcataa caaccatctc gattagggat aacagggtaa
3301 ttcttgtcat cgtcatcctt gtagtccatg gtggcctctc cttcgagtgc aaaaagaaca
3361 agtagcttgt attctatagt gtcacctaaa tgctagccta tagtgagtcg tattaagtac
3421 tctagcctta agagctgtaa ttgaactggg agtggacacc tgtggagaga aaggcaaagt
3481 ggatgtcagt aagaccaata ggtgcctatc agaaacgcaa gagtcttctc tgtctcgaca
3541 agcccagttt ctattggtct ccttaaacct gtcttgtaac cttgatactt acctgcccag
3601 tgcctcacga ccaactttcg ctaccttagg accgttatag ttactgcagc gctagcataa
3661 cttcgtataa agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat
3721 tctttagaat ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag ttatgaattc
3781 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
3841 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
3901 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
3961 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
4021 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
4081 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
4141 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
4201 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc
4261 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
4321 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
4381 tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct
4441 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
4501 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
4561 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
4621 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
4681 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
4741 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
4801 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
4861 tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg
4921 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
4981 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
5041 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
5101 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
5161 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
5221 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
5281 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
5341 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
5401 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
5461 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
5521 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
5581 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
5641 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
5701 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
5761 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
5821 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
5881 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
5941 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
6001 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
6061 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
6121 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
6181 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
6241 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
6301 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
6361 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
6421 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
6481 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
6541 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
6601 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
6661 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
6721 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
6781 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
6841 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
6901 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
6961 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
7021 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
7081 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
7141 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
7201 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
7261 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
7321 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
7381 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
7441 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
7501 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
7561 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
7621 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
