| 质粒名称: | pAAV-hSyn-hM3D(Gq)-mCherry |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 50474 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 空载体大小: | 4818 bp |
| 完整质粒大小: | 7059 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: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | hM3D(Gq)-mCherry |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 2502 bp |
| 基因突变: | T2A, S13L, A92T, V104T, V114I, E161K, Y193K, F198Y, D202T, I203V, Y258F, A283V, K288T, N290Y relative to RpBphP2 |
| GenBank编号 | JN247409 |
| 启动子: | human Synapsin 1 |
| 标签/融合蛋白: | mCherry (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | Sal I(未破坏) |
| 3’克隆位点: | EcoR I(未破坏) |
| 5’测序引物 | TCGTGTCGTGCCTGAGAGCG |
| 3’测序引物 | GCATTAAAGCAGCGTATCCACATAGC |
|
文献引用方法 |
|
| 材料和方法部分: |
pAAV-hSyn-hM3D(Gq)-mCherry was a gift from Bryan Roth (Addgene plasmid # 50474 ; http://n2t.net/addgene:50474 ; RRID:Addgene_50474)
For viral preps, please replace (Addgene plasmid # 50474) in the above sentence with: (Addgene viral prep # 50474-AAV2), (Addgene viral prep # 50474-AAV5), (Addgene viral prep # 50474-AAV8), (Addgene viral prep # 50474-AAV9), or (Addgene viral prep # 50474-AAVrg)
|
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
-mCherry质粒图谱.png)
载体简介
载体序列
LOCUS pAAV-hSyn-hM3D(Gq)-mCherry 7059 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Gq-coupled hM3D DREADD fused with mCherry under the control of human
synapsin promoter .
ACCESSION .
VERSION .
KEYWORDS pAAV-hSyn-hM3D(Gq)-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7059)
AUTHORS Bryan Roth
TITLE Roth lab DREADDs
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7059)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7059
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 1..23
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(61..79)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 147..251
/gene="bla"
/label=AmpR promoter
CDS 252..1112
/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(470..489)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1283..1871
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1772..1791
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
repeat_region 1933..2062
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
promoter 2107..2554
/label=hSyn promoter
/note="human synapsin I promoter; confers neuron-specific
expression (Kugler et al., 2003)"
regulatory 4377..4386
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4383..5093
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(4528..4546)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 4796..4815
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 5118..5706
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5171..5191)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5589..5600)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 5738..6214
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(5937..5956)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 6254..6394
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 6254..6383
/label=AAV2 ITR
rep_origin 6469..6924
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6556..6575)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6766..6787
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6941..6960)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
61 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
121 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
181 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
241 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
301 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
361 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
421 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
481 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
541 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
601 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
661 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
721 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
781 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
841 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
901 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
961 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
1021 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
1081 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
1141 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
1201 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
1261 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
1321 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
1381 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
1441 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
1501 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
1561 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
1621 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
1681 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
1741 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
1801 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
1861 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
1921 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt
1981 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
2041 aactccatca ctaggggttc ctgcggccgc acgcgtgtgt ctagactgca gagggccctg
2101 cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg cctacctgac
2161 gaccgacccc gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct
2221 atcagagagg gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc
2281 accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga
2341 aggcgcgctg acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg
2401 cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca
2461 cgggcgcgac catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg
2521 cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaga aggtaccgga tcctctagag
2581 tcgacgccac catgaccttg cacaataaca gtacaacctc gcctttgttt ccaaacatca
2641 gctcctcctg gatacacagc ccctccgatg cagggctgcc cccgggaacc gtcactcatt
2701 tcggcagcta caatgtttct cgagcagctg gcaatttctc ctctccagac ggtaccaccg
2761 atgaccctct gggaggtcat accgtctggc aagtggtctt catcgctttc ttaacgggca
2821 tcctggcctt ggtgaccatc atcggcaaca tcctggtaat tgtgtcattt aaggtcaaca
2881 agcagctgaa gacggtcaac aactacttcc tcttaagcct ggcctgtgcc gatctgatta
2941 tcggggtcat ttcaatgaat ctgtttacga cctacatcat catgaatcga tgggccttag
3001 ggaacttggc ctgtgacctc tggcttgcca ttgactgcgt agccagcaat gcctctgtta
3061 tgaatcttct ggtcatcagc tttgacagat acttttccat cacgaggccg ctcacgtacc
3121 gagccaaacg aacaacaaag agagccggtg tgatgatcgg tctggcttgg gtcatctcct
3181 ttgtcctttg ggctcctgcc atcttgttct ggcaatactt tgttggaaag agaactgtgc
3241 ctccgggaga gtgcttcatt cagttcctca gtgagcccac cattactttt ggcacagcca
3301 tcgctggttt ttatatgcct gtcaccatta tgactatttt atactggagg atctataagg
3361 aaactgaaaa gcgtaccaaa gagcttgctg gcctgcaagc ctctgggaca gaggcagaga
3421 cagaaaactt tgtccacccc acgggcagtt ctcgaagctg cagcagttac gaacttcaac
3481 agcaaagcat gaaacgctcc aacaggagga agtatggccg ctgccacttc tggttcacaa
3541 ccaagagctg gaaacccagc tccgagcaga tggaccaaga ccacagcagc agtgacagtt
3601 ggaacaacaa tgatgctgct gcctccctgg agaactccgc ctcctccgac gaggaggaca
3661 ttggctccga gacgagagcc atctactcca tcgtgctcaa gcttccgggt cacagcacca
3721 tcctcaactc caccaagtta ccctcatcgg acaacctgca ggtgcctgag gaggagctgg
3781 ggatggtgga cttggagagg aaagccgaca agctgcaggc ccagaagagc gtggacgatg
3841 gaggcagttt tccaaaaagc ttctccaagc ttcccatcca gctagagtca gccgtggaca
3901 cagctaagac ttctgacgtc aactcctcag tgggtaagag cacggccact ctacctctgt
3961 ccttcaagga agccactctg gccaagaggt ttgctctgaa gaccagaagt cagatcacta
4021 agcggaaaag gatgtccctg gtcaaggaga agaaagcggc ccagaccctc agtgcgatct
4081 tgcttgcctt catcatcact tggaccccat acaacatcat ggttctggtg aacacctttt
4141 gtgacagctg catacccaaa accttttgga atctgggcta ctggctgtgc tacatcaaca
4201 gcaccgtgaa ccccgtgtgc tatgctctgt gcaacaaaac attcagaacc actttcaaga
4261 tgctgctgct gtgccagtgt gacaaaaaaa agaggcgcaa gcagcagtac cagcagagac
4321 agtcggtcat ttttcacaag cgcgcacccg agcaggcctt gaaggatccc ccggtcgcca
4381 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
4441 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
4501 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
4561 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
4621 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
4681 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
4741 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
4801 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
4861 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
4921 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
4981 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
5041 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taagaattcg
5101 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
5161 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
5221 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
5281 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
5341 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
5401 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
5461 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
5521 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
5581 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
5641 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
5701 ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc
5761 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
5821 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
5881 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
5941 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
6001 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
6061 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
6121 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
6181 acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg
6241 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
6301 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
6361 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
6421 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
6481 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
6541 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
6601 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
6661 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
6721 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
6781 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
6841 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
6901 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
6961 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
7021 tgctcccggc atccgcttac agacaagctg tgaccgtct
//
