| 质粒名称: | pAAV_hsyn_NES-his-CaMPARI2-F391W-L398V |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 119723 |
| 存储实验室: | BThomas Oertner |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体出品公司: | Stratagene |
| 空载体大小: | 4267 bp |
| 完整质粒大小: | 5575 bp |
| 载体类型: | 哺乳细胞表达 |
| 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1: | CaMPARI2 |
| 物种来源: | Synthetic |
| 插入大小: | 1308bp |
| 基因突变: | F391W, L398V |
| 启动子: | hSyn1 |
| 标签/融合蛋白: | NES-his (N terminal on insert) |
| 克隆方法: | 未知 |
| 5’测序引物 | syn-sense2 GACTCAGCGCTGCCTCAGTCTG |
| 3’测序引物 | P5E gtaatccagaggttgattatcg |
| 插入基因名称2: | CaMPARI2 |
| 物种来源: | Synthetic |
| 插入大小: | 1308bp |
| 克隆方法: | 未知 |
| 5’测序引物 | syn-sense2 GACTCAGCGCTGCCTCAGTCTG |
| 3’测序引物 | P5E gtaatccagaggttgattatcg |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV_hsyn_NES-his-CaMPARI2-F391W-L398V was a gift from Thomas Oertner (Addgene plasmid # 119723 ; http://n2t.net/addgene:119723 ; RRID:Addgene_119723) |
| 参考文献部分: | Improved methods for marking active neuron populations. Moeyaert B, Holt G, Madangopal R, Perez-Alvarez A, Fearey BC, Trojanowski NF, Ledderose J, Zolnik TA, Das A, Patel D, Brown TA, Sachdev RNS, Eickholt BJ, Larkum ME, Turrigiano GG, Dana H, Gee CE, Oertner TG, Hope BT, Schreiter ER. Nat Commun. 2018 Oct 25;9(1):4440. doi: 10.1038/s41467-018-06935-2. 10.1038/s41467-018-06935-2 PubMed 30361563 |
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
载体简介
载体序列
LOCUS pAAV_hsyn_NES-his-CaMPARI2-F391W-L398V 5426 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION AAV vector expressing CaMPARI2_F391W-L398V (Kd = 174nM), a
photoconvertible fluorescent protein-based calcium integrator, in
neurons.
ACCESSION .
VERSION .
KEYWORDS pAAV_hsyn_NES-his-CaMPARI2-F391W-L398V
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5426)
AUTHORS Moeyaert B, Holt G, Madangopal R, Perez-Alvarez A, Fearey BC,
Trojanowski NF, Ledderose J, Zolnik TA, Das A, Patel D, Brown TA,
Sachdev RNS, Eickholt BJ, Larkum ME, Turrigiano GG, Dana H, Gee CE,
Oertner TG, Hope BT, Schreiter ER
TITLE Improved methods for marking active neuron populations.
JOURNAL Nat Commun. 2018 Oct 25;9(1):4440. doi: 10.1038/s41467-018-06935-2.
PUBMED 30361563
REFERENCE 2 (bases 1 to 5426)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5426
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 138..267
/label=AAV2 ITR
promoter 349..796
/label=hSyn promoter
/note="human synapsin I promoter; confers neuron-specific
expression (Kugler et al., 2003)"
CDS 893..910
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 917..1357
/codon_start=1
/product="calcium-binding protein"
/label=calmodulin
/translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
CDS 2078..2134
/codon_start=1
/product="calmodulin-binding peptide from avian smooth
muscle myosin light chain kinase"
/label=calmodulin-binding peptide
/translation="RRKWNKTGHAVRAIGRLSS"
misc_feature 2150..2738
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2203..2223)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2621..2632)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(2741..2757)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2742..2758)
/label=pBluescriptKS
/note="For pBluescript vector"
polyA_signal 2787..2908
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2798..2817)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2852..2871
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(3116..3133)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3287..3875)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3367..3386)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4046..4906)
/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 4669..4688
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4907..5011)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5037..66)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5174..5195)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5386..5405
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
61 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
121 cttcgctatt acgccagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
181 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
241 tggccaactc catcactagg ggttccttgt agttaatgat taacccgcca tgctacttat
301 ctacgtagcc atgctctagg aagatctctg cagagggccc tgcgtatgag tgcaagtggg
361 ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc ccgacccact
421 ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga gggggagggg
481 aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga cagtgccttc
541 gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc tgacgtcact
601 cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg
661 cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg accatctgcg
721 ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc
781 gtgcctgaga gcgcagtcga attcaagctg ctagcaagga tccgccacca tgctgcagaa
841 cgagcttgct cttaagttgg ctggacttga tattaacaag actggaggtt ctcatcatca
901 ccaccaccat ggatccgacc aactgactga agagcagatc gcagaattta aagaggcttt
961 ctccctattt gacaaggacg gggatgggac aataacaacc aaggagctgg ggacggtgat
1021 gcggtctctg gggcagaacc ccacagaagc agagctgcag gacatgatca atgaagtaga
1081 tgccgacggt gacggcacaa tcgacttccc tgagttcctg acaatgatgg caagaaaaat
1141 gaaagacaca gacagtgaag aagaaattag agaagcgttc cgtgtgtttg ataaggatgg
1201 caatggctac atcagtgcag cagagcttcg ccacgtgatg acaaaccttg gagagaagtt
1261 aacagatgaa gaggttgatg aaatgatcag ggaagcagac atcgatgggg atggtcaggt
1321 aaactacgaa gagtttgtag tgatgatgac agcgaagctc gagtgcgaga aaatctatgt
1381 gcgtgatgga gtgctgacgg gtgatattca tatgatcttg ttgcttgaag gaaatgccca
1441 ttaccgatgt gacttcagaa ctacttacaa agctaaggag aagggtgtca agttaccagg
1501 cgtgcactat gtggaccaca ccattgagat tttaagccat gacaaagatt acaacaaggt
1561 taagatctat gagtatgctg ttgctcattc tggattgcct gacaatgcca gacgaggcgg
1621 taccggcgga tccatggtga gtgcgattaa gccagacatg aagatcaaac tgcgtatgga
1681 aggcaacgta aacgggcacc actttgtgat cgacggagat ggtacaggca agccttatga
1741 gggaaaacag accatggatc ttgaagtcaa agagggcgga cctctgcctt ttgcctttga
1801 tatcctgacc actgcattcc attacggcaa cagggtattc gtgaaatatc cagacaacat
1861 acaagactat tttaagcagt cgtttcctaa ggggtattcg tgggaacgaa gcatgacttt
1921 cgaagacggg ggcatttgct atgccagaag cgacataaca atggaagggg acactttcta
1981 taataaagtt cgattttatg gtaccaactt tcccgccaat ggtccagtta tgcagaagaa
2041 gacgctgaaa tggatgccga gctggacgcg ttcatcacgt cgtaagtgga ataagacagg
2101 tcacgcagtc agagctatag gtcggctgag ctcataaaag cttatcgata atcaacctct
2161 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
2221 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
2281 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
2341 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
2401 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
2461 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
2521 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
2581 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
2641 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
2701 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgacc
2761 cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
2821 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
2881 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
2941 ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
3001 aatcgataag gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca
3061 ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
3121 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
3181 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
3241 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
3301 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
3361 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
3421 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
3481 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
3541 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
3601 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
3661 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
3721 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
3781 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
3841 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
3901 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
3961 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
4021 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
4081 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
4141 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
4201 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
4261 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
4321 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
4381 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
4441 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
4501 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
4561 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
4621 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
4681 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
4741 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
4801 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
4861 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
4921 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
4981 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
5041 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
5101 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
5161 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
5221 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
5281 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
5341 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
5401 aggagcgggc gctagggcgc tggcaa
//
