| 质粒名称: | pAAV-CAG-mNeonGreen |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 99134 |
| 存储实验室: | Viviana Gradinaru |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 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: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | mNeonGreen |
| 物种来源: | -- |
| 插入大小: | 3296 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | CAG |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | KpnI(未破坏) |
| 3’克隆位点: | EcoRI/HindIII (未破坏) |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-CAG-mNeonGreen was a gift from Viviana Gradinaru (Addgene plasmid # 99134 ; http://n2t.net/addgene:99134 ; RRID:Addgene_99134) |
| 参考文献部分: | Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL, Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, Gradinaru V. Nat Neurosci. 2017 Aug;20(8):1172-1179. doi: 10.1038/nn.4593. Epub 2017 Jun 26. 10.1038/nn.4593 PubMed 28671695 |
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-CAG-mNeonGreen 5882 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION An AAV genome that ubiquitously expresses the fluorescent protein
mNeonGreen from the CAG promoter.
ACCESSION .
VERSION .
KEYWORDS pAAV-CAG-mNeonGreen
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5882)
AUTHORS Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL,
Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, Gradinaru V
TITLE Engineered AAVs for efficient noninvasive gene delivery to the
central and peripheral nervous systems.
JOURNAL Nat Neurosci. 2017 Aug;20(8):1172-1179. doi: 10.1038/nn.4593. Epub
2017 Jun 26.
PUBMED 28671695
REFERENCE 2 (bases 1 to 5882)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5882
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..130
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
enhancer 210..513
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 515..792
/label=chicken beta-actin promoter
primer_bind 1122..1144
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1206..1225
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 1268..1277
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1274..1984
/codon_start=1
/product="bright monomeric yellow-green fluorescent protein
derived from LanYFP (Shaner et al., 2013)"
/label=mNeonGreen
/translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
KAFTDVMGMDELYK"
regulatory 1529..1538
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
misc_feature 2009..2597
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2062..2082)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2480..2491)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 2629..3105
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(2828..2847)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 3145..3285
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3145..3274
/label=AAV2 ITR
rep_origin 3360..3815
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3447..3466)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3657..3678
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3832..3851)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3951..3973
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4011..4029)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4097..4201
/gene="bla"
/label=AmpR promoter
CDS 4202..5062
/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(4420..4439)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5233..5821
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5722..5741
/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 gcgtaacact agttattaat agtaatcaat tacggggtca
181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
481 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
541 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
601 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg
661 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
721 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
781 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
841 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
901 ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg
961 tggctgcgtg aaagccttga ggggctccgg gagcgccggc aggaaggaaa tgggcgggga
1021 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
1081 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
1141 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
1201 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgcggcccaa
1261 cggtaccgcc accatggtga gcaagggcga ggaggataac atggcctctc tcccagcgac
1321 acatgagtta cacatctttg gctccatcaa cggtgtggac tttgacatgg tgggtcaggg
1381 caccggcaat ccaaatgatg gttatgagga gttaaacctg aagtccacca agggtgacct
1441 ccagttctcc ccctggattc tggtccctca tatcgggtat ggcttccatc agtacctgcc
1501 ctaccctgac gggatgtcgc ctttccaggc cgccatggta gatggctccg gataccaagt
1561 ccatcgcaca atgcagtttg aagatggtgc ctcccttact gttaactacc gctacaccta
1621 cgagggaagc cacatcaaag gagaggccca ggtgaagggg actggtttcc ctgctgacgg
1681 tcctgtgatg accaactcgc tgaccgctgc ggactggtgc aggtcgaaga agacttaccc
1741 caacgacaaa accatcatca gtacctttaa gtggagttac accactggaa atggcaagcg
1801 ctaccggagc actgcgcgga ccacctacac ctttgccaag ccaatggcgg ctaactatct
1861 gaagaaccag ccgatgtacg tgttccgtaa gacggagctc aagcactcca agaccgagct
1921 caacttcaag gagtggcaaa aggcctttac cgatgtgatg ggcatggacg agctgtacaa
1981 gtgagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
2041 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
2101 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
2161 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
2221 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
2281 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
2341 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
2401 gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac
2461 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
2521 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
2581 ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc
2641 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
2701 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
2761 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
2821 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
2881 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
2941 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
3001 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
3061 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta
3121 accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
3181 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
3241 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
3301 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
3361 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
3421 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
3481 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
3541 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
3601 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
3661 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
3721 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
3781 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
3841 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
3901 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
3961 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
4021 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
4081 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
4141 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
4201 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
4261 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
4321 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
4381 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
4441 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
4501 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
4561 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
4621 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
4681 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
4741 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
4801 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
4861 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
4921 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
4981 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
5041 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
5101 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
5161 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
5221 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
5281 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
5341 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
5401 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
5461 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
5521 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
5581 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
5641 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
5701 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
5761 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
5821 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
5881 gt
//
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