| 质粒名称: | pAAV-CaMKIIa-hChR2(C128S/D156A)-mCherry |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 35502 |
| 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: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5385 bp |
| 完整质粒大小: | -- |
| 载体修饰: | 添加CaMKIIa促进剂和WPRE |
| 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(C128S/D156A)-mCherry,SSFO-mCherry |
| 物种来源: | 莱茵衣藻 |
| 插入大小: | 1646 bp |
| 基因突变: | C128S and D156A |
| GenBank编号 | -- |
| 启动子 | CaMKIIa |
| 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 (未破坏) |
| 3' 克隆位点: | EcoRI(未破坏) |
| 5' 测序引物 | AGTCCTGCAGTATTGTGTAT |
| 3' 测序引物 | GCAATAGCATGATACAAAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-CaMKIIa-hChR2(C128S/D156A)-mCherry was a gift from Karl Deisseroth (Addgene plasmid # 35502 ; http://n2t.net/addgene:35502 ; RRID:Addgene_35502) |
| 参考文献部分: | Neocortical excitation/inhibition balance in information processing and social dysfunction. Yizhar O, Fenno LE, Prigge M, Schneider F, Davidson TJ, O'Shea DJ, Sohal VS, Goshen I, Finkelstein J, Paz JT, Stehfest K, Fudim R, Ramakrishnan C, Huguenard JR, Hegemann P, Deisseroth K. Nature. 2011 Jul 27;477(7363):171-8. doi: 10.1038/nature10360. 10.1038/nature10360 PubMed 21796121 |
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-CaMKIIa-hChR2(C128S/D156A)-mCherry 7031 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pAAV-CaMKIIa-hChR2(C128S/D156A)-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7031)
AUTHORS Yizhar O, Fenno LE, Prigge M, Schneider F, Davidson TJ, O'Shea DJ,
Sohal VS, Goshen I, Finkelstein J, Paz JT, Stehfest K, Fudim R,
Ramakrishnan C, Huguenard JR, Hegemann P, Deisseroth K
TITLE Neocortical excitation/inhibition balance in information processing
and social dysfunction.
JOURNAL Nature. 2011 Jul 27;477(7363):171-8. doi: 10.1038/nature10360.
PUBMED 21796121
REFERENCE 2 (bases 1 to 7031)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7031
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..141
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 12..141
/label=AAV2 ITR
regulatory 1481..1490
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2426..3133
/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(2568..2586)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2836..2855
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 3158..3746
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3211..3231)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3629..3640)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3778..4254
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3977..3996)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 4294..4434
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 4294..4423
/label=AAV2 ITR
rep_origin 4509..4964
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4596..4615)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4806..4827
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4981..5000)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5100..5122
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5160..5178)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5246..5350
/gene="bla"
/label=AmpR promoter
CDS 5351..6211
/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(5569..5588)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6382..6970
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6871..6890
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
121 actccatcac taggggttcc tgcggccgca cgcgtttaac attatggcct taggtcactt
181 catctccatg gggttcttct tctgattttc tagaaaatga gatgggggtg cagagagctt
241 cctcagtgac ctgcccaggg tcacatcaga aatgtcagag ctagaacttg aactcagatt
301 actaatctta aattccatgc cttgggggca tgcaagtacg atatacagaa ggagtgaact
361 cattagggca gatgaccaat gagtttagga aagaagagtc cagggcaggg tacatctaca
421 ccacccgccc agccctgggt gagtccagcc acgttcacct cattatagtt gcctctctcc
481 agtcctacct tgacgggaag cacaagcaga aactgggaca ggagccccag gagaccaaat
541 cttcatggtc cctctgggag gatgggtggg gagagctgtg gcagaggcct caggaggggc
601 cctgctgctc agtggtgaca gataggggtg agaaagcaga cagagtcatt ccgtcagcat
661 tctgggtctg tttggtactt cttctcacgc taaggtggcg gtgtgatatg cacaatggct
721 aaaaagcagg gagagctgga aagaaacaag gacagagaca gaggccaagt caaccagacc
781 aattcccaga ggaagcaaag aaaccattac agagactaca agggggaagg gaaggagaga
841 tgaattagct tcccctgtaa accttagaac ccagctgttg ccagggcaac ggggcaatac
901 ctgtctcttc agaggagatg aagttgccag ggtaactaca tcctgtcttt ctcaaggacc
961 atcccagaat gtggcaccca ctagccgtta ccatagcaac tgcctctttg ccccacttaa
1021 tcccatcccg tctgttaaaa gggccctata gttggaggtg ggggaggtag gaagagcgat
1081 gatcacttgt ggactaagtt tgttcgcatc cccttctcca accccctcag tacatcaccc
1141 tgggggaaca gggtccactt gctcctgggc ccacacagtc ctgcagtatt gtgtatataa
1201 ggccagggca aagaggagca ggttttaaag tgaaaggcag gcaggtgttg gggaggcagt
1261 taccggggca acgggaacag ggcgtttcgg aggtggttgc catggggacc tggatgctga
1321 cgaaggctcg cgaggctgtg agcagccaca gtgccctgct cagaagcccc aagctcgtca
1381 gtcaagccgg ttctccgttt gcactcagga gcacgggcag gcgagtggcc cctagttctg
1441 ggggcagctc tagagcggta ccggatcccc gggtaccggt gccaccatgg actatggcgg
1501 cgctttgtct gccgtcggac gcgaactttt gttcgttact aatcctgtgg tggtgaacgg
1561 gtccgtcctg gtccctgagg atcaatgtta ctgtgccgga tggattgaat ctcgcggcac
1621 gaacggcgct cagaccgcgt caaatgtcct gcagtggctt gcagcaggat tcagcatttt
1681 gctgctgatg ttctatgcct accaaacctg gaaatctaca tgcggctggg aggagatcta
1741 tgtgtgcgcc attgaaatgg ttaaggtgat tctcgagttc ttttttgagt ttaagaatcc
1801 ctctatgctc taccttgcca caggacaccg ggtgcagtgg ctgcgctatg cagagtggct
1861 gctcacttct cctgtcatcc ttatccacct gagcaacctc accggcctga gcaacgacta
1921 cagcaggaga accatgggac tccttgtctc agccatcggg actatcgtgt ggggggctac
1981 cagcgccatg gcaaccggct atgttaaagt catcttcttt tgtcttggat tgtgctatgg
2041 cgcgaacaca ttttttcacg ccgccaaagc atatatcgag ggttatcata ctgtgccaaa
2101 gggtcggtgc cgccaggtcg tgaccggcat ggcatggctg tttttcgtga gctggggtat
2161 gttcccaatt ctcttcattt tggggcccga aggttttggc gtcctgagcg tctatggctc
2221 caccgtaggt cacacgatta ttgatctgat gagtaaaaat tgttgggggt tgttgggaca
2281 ctacctgcgc gtcctgatcc acgagcacat attgattcac ggagatatcc gcaaaaccac
2341 caaactgaac atcggcggaa cggagatcga ggtcgagact ctcgtcgaag acgaagccga
2401 ggccggagcc gtgccagcgg ccgccgtgag caagggcgag gaggataaca tggccatcat
2461 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
2521 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
2581 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
2641 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
2701 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
2761 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
2821 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
2881 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
2941 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
3001 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
3061 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
3121 gctgtacaag taagaattcg atatcaagct tatcgataat caacctctgg attacaaaat
3181 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
3241 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
3301 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
3361 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
3421 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
3481 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
3541 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
3601 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
3661 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
3721 gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg
3781 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
3841 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
3901 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
3961 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
4021 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
4081 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
4141 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
4201 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
4261 ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
4321 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
4381 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
4441 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
4501 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
4561 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
4621 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
4681 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
4741 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
4801 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt
4861 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
4921 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
4981 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
5041 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
5101 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
5161 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
5221 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
5281 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
5341 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
5401 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
5461 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
5521 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
5581 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
5641 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
5701 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
5761 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
5821 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
5881 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
5941 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
6001 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
6061 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
6121 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
6181 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
6241 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
6301 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
6361 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
6421 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
6481 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
6541 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
6601 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
6661 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
6721 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
6781 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
6841 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
6901 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
6961 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
7021 tgctcacatg t
//
