| 质粒名称: | pAAV-hSyn-hChR2(H134R)-mCherry |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 26976 |
| 存储实验室: | Karl Deisseroth |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4568 bp |
| 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: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | hChR2(H134R),hChR2 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1647 bp |
| 基因突变: | H134R |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | mCherry（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI, SalI, KpnI(未破坏) |
| 3' 克隆位点: | EcoRI, HinDIII (未破坏) |
| 5' 测序引物 | ccacgcgaggcgcgagatag |
| 3' 测序引物 | GCAATAGCATGATACAAAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-hSyn-hChR2(H134R)-mCherry was a gift from Karl Deisseroth (Addgene plasmid # 26976 ; http://n2t.net/addgene:26976 ; RRID:Addgene_26976) |
| 23770755 | |
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-hChR2(H134R)-mCherry 6204 bp ds-DNA circular SYN 01-OCT-2020
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pAAV-hSyn-hChR2(H134R)-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6204)
TITLE hChR2 plasmids
REFERENCE 2 (bases 1 to 6204)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 1, 2020 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6204
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..448
/label=hSyn promoter
/note="human synapsin I promoter; confers neuron-specific
expression (Kugler et al., 2003)"
regulatory 480..489
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1425..2132
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
GRHSTGGMDELYK"
primer_bind complement(1567..1585)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1835..1854
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 2157..2745
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2210..2230)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2628..2639)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 2777..3253
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(2976..2995)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 3293..3433
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3293..3422
/label=AAV2 ITR
rep_origin 3508..3963
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3595..3614)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3805..3826
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3980..3999)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4099..4121
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4159..4177)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4245..4349
/gene="bla"
/label=AmpR promoter
CDS 4350..5210
/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(4568..4587)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5381..5969
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5870..5889
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
repeat_region 6031..6160
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
421 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggtac cggatcctct agagtcgacg
481 ccaccatgga ctatggcggc gctttgtctg ccgtcggacg cgaacttttg ttcgttacta
541 atcctgtggt ggtgaacggg tccgtcctgg tccctgagga tcaatgttac tgtgccggat
601 ggattgaatc tcgcggcacg aacggcgctc agaccgcgtc aaatgtcctg cagtggcttg
661 cagcaggatt cagcattttg ctgctgatgt tctatgccta ccaaacctgg aaatctacat
721 gcggctggga ggagatctat gtgtgcgcca ttgaaatggt taaggtgatt ctcgagttct
781 tttttgagtt taagaatccc tctatgctct accttgccac aggacaccgg gtgcagtggc
841 tgcgctatgc agagtggctg ctcacttgtc ctgtcatcct tatccgcctg agcaacctca
901 ccggcctgag caacgactac agcaggagaa ccatgggact ccttgtctca gacatcggga
961 ctatcgtgtg gggggctacc agcgccatgg caaccggcta tgttaaagtc atcttctttt
1021 gtcttggatt gtgctatggc gcgaacacat tttttcacgc cgccaaagca tatatcgagg
1081 gttatcatac tgtgccaaag ggtcggtgcc gccaggtcgt gaccggcatg gcatggctgt
1141 ttttcgtgag ctggggtatg ttcccaattc tcttcatttt ggggcccgaa ggttttggcg
1201 tcctgagcgt ctatggctcc accgtaggtc acacgattat tgatctgatg agtaaaaatt
1261 gttgggggtt gttgggacac tacctgcgcg tcctgatcca cgagcacata ttgattcacg
1321 gagatatccg caaaaccacc aaactgaaca tcggcggaac ggagatcgag gtcgagactc
1381 tcgtcgaaga cgaagccgag gccggagccg tgccagcggc cgccgtgagc aagggcgagg
1441 aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
1501 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
1561 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt
1621 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
1681 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
1741 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
1801 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
1861 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
1921 agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca
1981 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca
2041 cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact
2101 ccaccggcgg catggacgag ctgtacaagt aagaattcga tatcaagctt atcgataatc
2161 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
2221 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
2281 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
2341 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
2401 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
2461 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
2521 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
2581 atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
2641 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
2701 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc
2761 tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
2821 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
2881 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
2941 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
3001 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
3061 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
3121 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
3181 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
3241 cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc
3301 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
3361 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
3421 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
3481 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
3541 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
3601 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
3661 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
3721 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
3781 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
3841 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
3901 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
3961 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
4021 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
4081 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
4141 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
4201 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
4261 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
4321 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
4381 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
4441 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
4501 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
4561 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
4621 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
4681 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
4741 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
4801 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
4861 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
4921 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
4981 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
5041 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
5101 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
5161 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
5221 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
5281 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
5341 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
5401 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
5461 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
5521 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
5581 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
5641 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
5701 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
5761 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
5821 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
5881 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
5941 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
6001 tggccttttg ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca
6061 ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
6121 cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcac gcgtgtgtct
6181 agactgcaga gggccctgcg tatg
//
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