| 质粒名称: | pAAV_hsyn_NES-his-CaMPARI2-L398T-WPRE-SV40 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 101064 |
| 存储实验室: | Eric Schreiter |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 空载体大小: | 4267 bp |
| 完整质粒大小: | 5567 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: | CaMPARI2_L398T |
| 物种来源: | Synthetic |
| 插入大小: | 1446bp |
| 标签/融合蛋白: |
NES_his (N terminal on insert)
FLAG-HA-myc (C terminal on insert)
|
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | f.hsyn |
| 3’测序引物 | r.WPRE |
|
文献引用方法 |
|
| 材料和方法部分: |
pAAV_hsyn_NES-his-CaMPARI2-L398T-WPRE-SV40 was a gift from Eric Schreiter (Addgene plasmid # 101064 ; http://n2t.net/addgene:101064 ; RRID:Addgene_101064)
For viral preps, please replace (Addgene plasmid # 101064) in the above sentence with: (Addgene viral prep # 101064-AAV1)
|
| 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-L398T-WPRE-SV40 5567 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION AAV vector expressing CaMPARI2_L398T (Kd = 825nM), a
photoconvertible fluorescent protein-based calcium integrator, in
neurons.
ACCESSION .
VERSION .
KEYWORDS pAAV_hsyn_NES-his-CaMPARI2-L398T-WPRE-SV40
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5567)
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 5567)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5567
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 9..456
/label=hSyn promoter
/note="human synapsin I promoter; confers neuron-specific
expression (Kugler et al., 2003)"
CDS 553..570
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 577..1017
/codon_start=1
/product="calcium-binding protein"
/label=calmodulin
/translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
CDS 1813..1836
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 1855..1881
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1900..1929
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
misc_feature 1951..2539
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2004..2024)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2422..2433)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(2542..2558)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2543..2559)
/label=pBluescriptKS
/note="For pBluescript vector"
polyA_signal 2588..2709
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2599..2618)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2653..2672
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(2917..2934)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3088..3676)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3168..3187)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3847..4707)
/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 4470..4489
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4708..4812)
/gene="bla"
/label=AmpR promoter
rep_origin complement(4838..5293)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4975..4996)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5187..5206
/label=F1ori-R
/note="F1 origin, reverse primer"
repeat_region 5365..5494
/label=AAV2 ITR
ORIGIN
1 tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg
61 acgaccgacc ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc
121 ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca
181 gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact
241 gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt
301 cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg
361 cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg
421 ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga attcaagctg ctagcaagga
481 tccgccacca tgctgcagaa cgagcttgct cttaagttgg ctggacttga tattaacaag
541 actggaggtt ctcatcatca ccaccaccat ggatccgacc aactgactga agagcagatc
601 gcagaattta aagaggcttt ctccctattt gacaaggacg gggatgggac aataacaacc
661 aaggagctgg ggacggtgat gcggtctctg gggcagaacc ccacagaagc agagctgcag
721 gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc tgagttcctg
781 acaatgatgg caagaaaaat gaaagacaca gacagtgaag aagaaattag agaagcgttc
841 cgtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg ccacgtgatg
901 acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag ggaagcagac
961 atcgatgggg atggtcaggt aaactacgaa gagtttgtag tgatgatgac agcgaagctc
1021 gagtgcgaga aaatctatgt gcgtgatgga gtgctgacgg gtgatattca tatgatcttg
1081 ttgcttgaag gaaatgccca ttaccgatgt gacttcagaa ctacttacaa agctaaggag
1141 aagggtgtca agttaccagg cgtgcactat gtggaccaca ccattgagat tttaagccat
1201 gacaaagatt acaacaaggt taagatctat gagtatgctg ttgctcattc tggattgcct
1261 gacaatgcca gacgaggcgg taccggcgga tccatggtga gtgcgattaa gccagacatg
1321 aagatcaaac tgcgtatgga aggcaacgta aacgggcacc actttgtgat cgacggagat
1381 ggtacaggca agccttatga gggaaaacag accatggatc ttgaagtcaa agagggcgga
1441 cctctgcctt ttgcctttga tatcctgacc actgcattcc attacggcaa cagggtattc
1501 gtgaaatatc cagacaacat acaagactat tttaagcagt cgtttcctaa ggggtattcg
1561 tgggaacgaa gcatgacttt cgaagacggg ggcatttgct atgccagaag cgacataaca
1621 atggaagggg acactttcta taataaagtt cgattttatg gtaccaactt tcccgccaat
1681 ggtccagtta tgcagaagaa gacgctgaaa tggatgccga gctggacgcg ttcatcacgt
1741 cgtaagttca ataagacagg tcacgcaacg agagctatag gtcggctgag ctcaggaggt
1801 tctggaggtt ctgattacaa ggacgacgat gacaagggag gttctggagg ttcttaccca
1861 tacgacgtac cagattacgc tggaggttct ggaggttctg agcagaagct gatctcagag
1921 gaggacctgc gtacgtaaaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
1981 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
2041 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
2101 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
2161 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
2221 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
2281 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
2341 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
2401 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
2461 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
2521 ctttgggccg cctccccgca tcgataccgt cgacctcgac ccgggcggcc gcttcgagca
2581 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
2641 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
2701 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
2761 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatcttcct
2821 agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct
2881 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgact cgctgcgctc
2941 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
3001 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
3061 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
3121 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
3181 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
3241 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
3301 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
3361 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
3421 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
3481 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
3541 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
3601 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
3661 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
3721 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
3781 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
3841 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
3901 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
3961 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
4021 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
4081 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
4141 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
4201 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
4261 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
4321 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
4381 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
4441 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
4501 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
4561 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
4621 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
4681 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
4741 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
4801 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
4861 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
4921 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
4981 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
5041 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
5101 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
5161 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
5221 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
5281 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
5341 cgggcctctt cgctattacg ccagctgcgc gctcgctcgc tcactgaggc cgcccgggca
5401 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
5461 gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa cccgccatgc
5521 tacttatcta cgtagccatg ctctaggaag atctctgcag agggccc
//
