| 质粒名称: | pAAV-hSyn-hM4D(Gi)-mCherry |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 50475 |
| 存储实验室: | Bryan Roth |
|
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: | 6726 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: | hM4D(Gi)-mCherry |
| 插入大小: | 2169bp |
| 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-hM4D(Gi)-mCherry was a gift from Bryan Roth (Addgene plasmid # 50475 ; http://n2t.net/addgene:50475 ; RRID:Addgene_50475)
For viral preps, please replace (Addgene plasmid # 50475) in the above sentence with: (Addgene viral prep # 50475-AAV2), (Addgene viral prep # 50475-AAV5), (Addgene viral prep # 50475-AAV8), (Addgene viral prep # 50475-AAV9), or (Addgene viral prep # 50475-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-hM4D(Gi)-mCherry 6726 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Gi-coupled hM4D DREADD fused with mCherry under the control of human
synapsin promoter..
ACCESSION .
VERSION .
KEYWORDS pAAV-hSyn-hM4D(Gi)-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6726)
AUTHORS Bryan Roth
TITLE Roth lab DREADDs
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6726)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6726
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..130
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
promoter 175..622
/label=hSyn promoter
/note="human synapsin I promoter; confers neuron-specific
expression (Kugler et al., 2003)"
regulatory 654..663
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 2112..2121
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2118..2828
/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(2263..2281)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2531..2550
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 2853..3441
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2906..2926)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3324..3335)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3473..3949
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3672..3691)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 3989..4129
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3989..4118
/label=AAV2 ITR
rep_origin 4204..4659
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4291..4310)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4501..4522
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4676..4695)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4795..4817
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4855..4873)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4941..5045
/gene="bla"
/label=AmpR promoter
CDS 5046..5906
/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(5264..5283)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6077..6665
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6566..6585
/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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gacgccacca
661 tggccaactt cacacctgtc aatggcagct cgggcaatca gtccgtgcgc ctggtcacgt
721 catcatccca caatcgctat gagacggtgg aaatggtctt cattgccaca gtgacaggct
781 ccctgagcct ggtgactgtc gtgggcaaca tcctggtgat gctgtccatc aaggtcaaca
841 ggcagctgca gacagtcaac aactacttcc tcttcagcct ggcgtgtgct gatctcatca
901 taggcgcctt ctccatgaac ctctacaccg tgtacatcat caagggctac tggcccctgg
961 gcgccgtggt ctgcgacctg tggctggccc tggactgcgt ggtgagcaac gcctccgtca
1021 tgaaccttct catcatcagc tttgaccgct acttctgcgt caccaagcct ctcacctacc
1081 ctgcccggcg caccaccaag atggcaggcc tcatgattgc tgctgcctgg gtactgtcct
1141 tcgtgctctg ggcgcctgcc atcttgttct ggcagtttgt ggtgggtaag cggacggtgc
1201 ccgacaacca gtgcttcatc cagttcctgt ccaacccagc agtgaccttt ggcacagcca
1261 ttgctggctt ctacctgcct gtggtcatca tgacggtgct gtacatccac atctccctgg
1321 ccagtcgcag ccgagtccac aagcaccggc ccgagggccc gaaggagaag aaagccaaga
1381 cgctggcctt cctcaagagc ccactaatga agcagagcgt caagaagccc ccgcccgggg
1441 aggccgcccg ggaggagctg cgcaatggca agctggagga ggcccccccg ccagcgctgc
1501 caccgccacc gcgccccgtg gctgataagg acacttccaa tgagtccagc tcaggcagtg
1561 ccacccagaa caccaaggaa cgcccagcca cagagctgtc caccacagag gccaccacgc
1621 ccgccatgcc cgcccctccc ctgcagccgc gggccctcaa cccagcctcc agatggtcca
1681 agatccagat tgtgacgaag cagacaggca atgagtgtgt gacagccatt gagattgtgc
1741 ctgccacgcc ggctggcatg cgccctgcgg ccaacgtggc ccgcaagttc gccagcatcg
1801 ctcgcaacca ggtgcgcaag aagcggcaga tggcggcccg ggagcgcaaa gtgacacgaa
1861 cgatctttgc cattctgctg gccttcatcc tcacctggac gccctacaac gtcatggtcc
1921 tggtgaacac cttctgccag agctgcatcc ctgacacggt gtggtccatt ggctactggc
1981 tctgctacgt caacagcacc atcaaccctg cctgctatgc tctgtgcaac gccaccttta
2041 aaaagacctt ccggcacctg ctgctgtgcc agtatcggaa catcggcact gccaggcggg
2101 atccaccggt cgccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
2161 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
2221 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
2281 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
2341 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
2401 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
2461 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
2521 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
2581 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
2641 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
2701 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
2761 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
2821 acaagtaaga attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
2881 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
2941 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
3001 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
3061 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
3121 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
3181 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
3241 cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg
3301 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
3361 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
3421 ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca
3481 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
3541 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
3601 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
3661 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
3721 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
3781 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
3841 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
3901 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta
3961 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
4021 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
4081 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
4141 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
4201 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
4261 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
4321 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
4381 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
4441 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
4501 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
4561 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
4621 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
4681 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
4741 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
4801 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
4861 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
4921 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
4981 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
5041 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
5101 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
5161 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
5221 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
5281 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
5341 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
5401 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
5461 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
5521 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
5581 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
5641 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
5701 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
5761 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
5821 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
5881 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
5941 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
6001 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
6061 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
6121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
6181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
6241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
6301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
6361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
6421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
6481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
6541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
6601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
6661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
6721 acatgt
//
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