| 质粒名称: | pAAV-hSyn-GRAB_DA1m |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 113049 |
| 存储实验室: | Yulong Li |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体出品公司: | Addgene |
| 空载体大小: | 4495 bp |
| 完整质粒大小: | 6329 bp |
| 载体类型: | AAV |
| 载体抗性: | 氨苄霉素 |
| 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: | GPCR activation based DA sensor GRAB_DA1m |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1851 bp |
| 启动子: | hSyn |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: |
pAAV-hSyn-GRAB_DA1m was a gift from Yulong Li (Addgene plasmid # 113049 ; http://n2t.net/addgene:113049 ; RRID:Addgene_113049)
For viral preps, please replace (Addgene plasmid # 113049) in the above sentence with: (Addgene viral prep # 113049-AAV9)
|
| 参考文献部分: | A Genetically Encoded Fluorescent Sensor Enables Rapid and Specific Detection of Dopamine in Flies, Fish, and Mice. Sun F, Zeng J, Jing M, Zhou J, Feng J, Owen SF, Luo Y, Li F, Wang H, Yamaguchi T, Yong Z, Gao Y, Peng W, Wang L, Zhang S, Du J, Lin D, Xu M, Kreitzer AC, Cui G, Li Y. Cell. 2018 Jul 12;174(2):481-496.e19. doi: 10.1016/j.cell.2018.06.042. 10.1016/j.cell.2018.06.042 PubMed 30007419 |
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-GRAB_DA1m 6329 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION express the genetically-encoded fluorescent dopamine(DA) sensor
GRAB_DA1m in neurons.
ACCESSION .
VERSION .
KEYWORDS pAAV-hSyn-GRAB_DA1m
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6329)
AUTHORS Sun F, Zeng J, Jing M, Zhou J, Feng J, Owen SF, Luo Y, Li F, Wang H,
Yamaguchi T, Yong Z, Gao Y, Peng W, Wang L, Zhang S, Du J, Lin D, Xu
M, Kreitzer AC, Cui G, Li Y
TITLE A Genetically Encoded Fluorescent Sensor Enables Rapid and Specific
Detection of Dopamine in Flies, Fish, and Mice.
JOURNAL Cell. 2018 Jul 12;174(2):481-496.e19. doi:
10.1016/j.cell.2018.06.042.
PUBMED 30007419
REFERENCE 2 (bases 1 to 6329)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6329
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..130
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
regulatory 575..584
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
sig_peptide 581..643
/product="leader sequence from mouse immunoglobulin kappa
light chain"
/label=Ig-kappa leader
protein_bind 644..668
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
primer_bind 1639..1660
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1766..1787)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2027..2046)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
misc_feature 2456..3044
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2509..2529)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2927..2938)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3076..3552
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3275..3294)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 3592..3732
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3592..3721
/label=AAV2 ITR
rep_origin 3807..4262
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3894..3913)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4104..4125
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4279..4298)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4398..4420
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4458..4476)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4544..4648
/gene="bla"
/label=AmpR promoter
CDS 4649..5509
/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(4867..4886)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5680..6268
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6169..6188
/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 gcgtatgagg cggggtgggg gtgcctacct gacgaccgac
181 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
241 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
301 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
361 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
421 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
481 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
541 ggagtcgtgt cgtgcctgag agcgcagtcg aaatgccacc atggagacag acacactcct
601 gctatgggta ctgctgctct gggttccagg ttccactggt gacacaagtt tgtacaaaaa
661 agttggcacc atggatccac tgaatctgtc ctggtatgat gatgatctgg agaggcagaa
721 ctggagccgg cccttcaacg ggtcagacgg gaaggcggac agaccccact acaactacta
781 tgccacactg ctcaccctgc tcatcgctgt catcgtcttc ggcaacgtgc tggtgtgcat
841 ggctgtgtcc cgcgagaagg cgctgcagac caccaccaac tacctgatcg tcagcctcgc
901 agtggccgac ctcctcgtcg ccacactggt catgccctgg gttgtctacc tggaggtggt
961 aggtgagtgg aaattcagca ggattcactg tgacatcttc gtcactctgg acgtcatgat
1021 gtgcacggcg agcatcctga acttgtgtgc catcagcatc gacaggtaca cagctgtggc
1081 catgcccatg ctgtacaata cgcgctacag ctccaagcgc cgggtcaccg tcatgatctc
1141 catcgtctgg gtcctgtcct tcaccatctc ctgcccactc ctcttcggac tcaataacgc
1201 agaccagaac gagtgcatca ttgccaaccc ggccttcgtg gtctactcct ccatcgtctc
1261 cttctacgtg cccttcattg tcaccctgct ggtctacatc aagatctaca ttgtcctccg
1321 cagacgccgc aagcgagtca acaccaaacg cagcagccga gctttcaggg cccacctgag
1381 ggctccacta aagggcaact gtactcaccc cgaggacatg aaactcggag gaaacgtcta
1441 tatcaaggcc gacaagcaga agaacggcat caaggcgaac ttccacatcc gccacaacat
1501 cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag aacaccccca tcggcgacgg
1561 ccccgtgctg ctgcccgaca accactacct gagcgtgcag tccaaacttt cgaaagaccc
1621 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
1681 cggcatggac gagctgtaca agggcggtac cggagggagc atggtgagaa agggcgagga
1741 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
1801 gttcagcgtg tccggcgagg gtgagggcga tgccacctac ggcaagctga ccctgaagtt
1861 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
1921 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
1981 cgccatgccc gaaggctaca tccaggagcg caccatcttc ttcaaggacg acggcaacta
2041 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
2101 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacggagg
2161 agcagcagca atgagccgta ggaagctctc ccagcagaag gagaagaaag ccactcagat
2221 gctcgccatt gttctcggcg tgttcatcat ctgctggctg cccttcttca tcacacacat
2281 cctgaacata cactgtgact gcaacatccc gcctgtcctg tacagcgcct tcacgtggct
2341 gggctatgtc aacagcgccg tgaaccccat catctacacc accttcaaca ttgagttccg
2401 caaggccttc ctgaagatcc tccactgctg agaattcgat atcaagctta tcgataatca
2461 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
2521 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
2581 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
2641 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
2701 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
2761 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
2821 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta
2881 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
2941 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
3001 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct
3061 gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
3121 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
3181 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
3241 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
3301 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
3361 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
3421 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
3481 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
3541 ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc
3601 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
3661 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
3721 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
3781 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
3841 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
3901 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
3961 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
4021 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
4081 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
4141 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
4201 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
4261 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
4321 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
4381 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
4441 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
4501 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
4561 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
4621 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
4681 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
4741 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
4801 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
4861 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
4921 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
4981 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
5041 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
5101 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
5161 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
5221 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
5281 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
5341 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
5401 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
5461 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
5521 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
5581 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
5641 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
5701 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
5761 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
5821 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
5881 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
5941 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
6001 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
6061 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
6121 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
6181 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
6241 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
6301 ggccttttgc tggccttttg ctcacatgt
//
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