| 质粒名称: | pAAV-hSyn-DA1h |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 113050 |
| 存储实验室: | Vladislav Verkhusha |
|
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: | 4495bp |
| 完整质粒大小: | 6338 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_DA1h |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1854 bp |
| 启动子: | hSyn |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | 未知 |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: |
pAAV-hSyn-GRAB_DA1h was a gift from Yulong Li (Addgene plasmid # 113050 ; http://n2t.net/addgene:113050 ; RRID:Addgene_113050)
For viral preps, please replace (Addgene plasmid # 113050) in the above sentence with: (Addgene viral prep # 113050-AAV9) or (Addgene viral prep # 113050-AAVrg)
|
| 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 |
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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-DA1h 6338 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION express the genetically-encoded fluorescent dopamine(DA) sensor
GRAB_DA1h in neurons.
ACCESSION .
VERSION .
KEYWORDS pAAV-hSyn-GRAB_DA1h
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6338)
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 6338)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6338
/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 581..590
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
sig_peptide 587..649
/product="leader sequence from mouse immunoglobulin kappa
light chain"
/label=Ig-kappa leader
protein_bind 650..674
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
primer_bind 1648..1669
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1775..1796)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2036..2055)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind complement(2518..2538)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2936..2947)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3085..3561
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3284..3303)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 3601..3741
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3601..3730
/label=AAV2 ITR
rep_origin 3816..4271
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3903..3922)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4113..4134
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4288..4307)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4407..4429
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4467..4485)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4553..4657
/gene="bla"
/label=AmpR promoter
CDS 4658..5518
/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(4876..4895)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5689..6277
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6178..6197
/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 agaaaccggt gccaccatgg agacagacac
601 actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca caagtttgta
661 caaaaaagtt ggcaccaccg gtgatccact gaatctgtcc tggtatgatg atgatctgga
721 gaggcagaac tggagccggc ccttcaacgg gtcagacggg aaggcggaca gaccccacta
781 caactactat gccacactgc tcaccctgct catcgctgtc atcgtcttcg gcaacgtgct
841 ggtgtgcatg gctgtgtccc gcgagaaggc gctgcagacc accaccaact acctgatcgt
901 cagcctcgca gtggccgacc tcctcgtcgc cacactggtc atgccctggg ttgtctacct
961 ggaggtggta ggtgagtgga aattcagcag gattcactgt gacatcttcg tcactctgga
1021 cgtcatgatg tgcacggcga gcatcctgaa cttgtgtgcc atcagcatcg acaggtacac
1081 agctgtggcc atgcccatgc tgtacaatac gcgctacagc tccaagcgcc gggtcaccgt
1141 catgatctcc atcgtctggg tcctgtcctt caccatctcc tgcccactcc tcttcggact
1201 caataacgca gaccagaacg agtgcatcat tgccaacccg gccttcgtgg tctactcctc
1261 catcgtctcc ttctacgtgc ccttcattgt catgctgctg gtctacatca agatctacat
1321 tgtcctccgc agacgccgca agcgagtcaa caccaaacgc agcagccgag ctttcagggc
1381 ccacctgagg gctccactaa agggcaactg tactcacccc gaggacatga aactcggagg
1441 aaacgtctat atcaaggccg acaagcagaa gaacggcatc aaggcgaact tccacatccg
1501 ccacaacatc gaggacggcg gcgtgcagct cgcctaccac taccagcaga acacccccat
1561 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcgtgcagt ccaaactttc
1621 gaaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
1681 gatcactctc ggcatggacg agctgtacaa gggcggtacc ggagggagca tggtgagcaa
1741 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
1801 cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg gcaagctgac
1861 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
1921 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
1981 cttcaagtcc gccatgcccg aaggctacat ccaggagcgc accatcttct tcaaggacga
2041 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
2101 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
2161 caacactgga gcagcagcaa tgagccgtag gaagctctcc cagcagaagg agaagaaagc
2221 cactcagatg ctcgccattg ttctcggcgt gttcatcatc tgctggctgc ccttcttcat
2281 cacacacatc ctgaacatac actgtgactg caacatcccg cctgtcctgt acagcgcctt
2341 cacgtggctg ggctatgtca acagcgccgt gaaccccatc atctacacca ccttcaacat
2401 tgagttccgc aaggccttcc tgaagatcct ccactgctga gaattcgata tcaagcttat
2461 cgatcatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
2521 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
2581 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
2641 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
2701 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
2761 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
2821 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
2881 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
2941 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
3001 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
3061 ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc
3121 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
3181 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
3241 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
3301 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
3361 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
3421 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
3481 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
3541 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc
3601 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
3661 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
3721 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
3781 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
3841 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
3901 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
3961 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
4021 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
4081 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
4141 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
4201 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
4261 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
4321 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
4381 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
4441 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
4501 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
4561 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
4621 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
4681 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
4741 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
4801 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
4861 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
4921 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
4981 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
5041 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
5101 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
5161 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
5221 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
5281 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
5341 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
5401 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
5461 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
5521 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
5581 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
5641 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
5701 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
5761 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
5821 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
5881 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
5941 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
6001 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
6061 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
6121 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
6181 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
6241 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
6301 acggttcctg gccttttgct ggccttttgc tcacatgt
//
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