| 质粒名称: | pAAV_hsyn_NES-his-CaMPARI2-H396K-WPRE-SV40 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 101062 |
| 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: | 4267bp |
| 完整质粒大小: | 5716bp |
| 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: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | CaMPARI2-H396K |
| 物种来源: | Synthetic |
| 插入大小: | 1446 bp |
| 启动子: | hsyn (synapsin-1) |
| 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’克隆位点: | AgeI(未破坏) |
| 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-H396K-WPRE-SV40 was a gift from Eric Schreiter (Addgene plasmid # 101062 ; http://n2t.net/addgene:101062 ; RRID:Addgene_101062) |
| 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-H396K-WPRE-SV40 5567 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION AAV vector expressing CaMPARI2_H396K (Kd = 360nM), a
photoconvertible fluorescent protein-based calcium integrator, in
neurons.
ACCESSION .
VERSION .
KEYWORDS pAAV_hsyn_NES-his-CaMPARI2-H396K-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 30..477
/label=hSyn promoter
/note="human synapsin I promoter; confers neuron-specific
expression (Kugler et al., 2003)"
CDS 574..591
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 598..1038
/codon_start=1
/product="calcium-binding protein"
/label=calmodulin
/translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
CDS 1834..1857
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 1876..1902
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1921..1950
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
misc_feature 1972..2560
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2025..2045)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2443..2454)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(2563..2579)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2564..2580)
/label=pBluescriptKS
/note="For pBluescript vector"
polyA_signal 2609..2730
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2620..2639)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2674..2693
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin complement(3109..3697)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3189..3208)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3868..4728)
/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 4491..4510
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4729..4833)
/gene="bla"
/label=AmpR promoter
rep_origin complement(4859..5314)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4996..5017)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5208..5227
/label=F1ori-R
/note="F1 origin, reverse primer"
repeat_region 5386..5515
/label=AAV2 ITR
ORIGIN
1 gaagatctct gcagagggcc ctgcgtatga gtgcaagtgg gttttaggac caggatgagg
61 cggggtgggg gtgcctacct gacgaccgac cccgacccac tggacaagca cccaaccccc
121 attccccaaa ttgcgcatcc cctatcagag agggggaggg gaaacaggat gcggcgaggc
181 gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg
241 cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact
301 ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac
361 gcgaggcgcg agataggggg gcacgggcgc gaccatctgc gctgcggcgc cggcgactca
421 gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg
481 aattcaagct gctagcaagg atccgccacc atgctgcaga acgagcttgc tcttaagttg
541 gctggacttg atattaacaa gactggaggt tctcatcatc accaccacca tggatccgac
601 caactgactg aagagcagat cgcagaattt aaagaggctt tctccctatt tgacaaggac
661 ggggatggga caataacaac caaggagctg gggacggtga tgcggtctct ggggcagaac
721 cccacagaag cagagctgca ggacatgatc aatgaagtag atgccgacgg tgacggcaca
781 atcgacttcc ctgagttcct gacaatgatg gcaagaaaaa tgaaagacac agacagtgaa
841 gaagaaatta gagaagcgtt ccgtgtgttt gataaggatg gcaatggcta catcagtgca
901 gcagagcttc gccacgtgat gacaaacctt ggagagaagt taacagatga agaggttgat
961 gaaatgatca gggaagcaga catcgatggg gatggtcagg taaactacga agagtttgta
1021 gtgatgatga cagcgaagct cgagtgcgag aaaatctatg tgcgtgatgg agtgctgacg
1081 ggtgatattc atatgatctt gttgcttgaa ggaaatgccc attaccgatg tgacttcaga
1141 actacttaca aagctaagga gaagggtgtc aagttaccag gcgtgcacta tgtggaccac
1201 accattgaga ttttaagcca tgacaaagat tacaacaagg ttaagatcta tgagtatgct
1261 gttgctcatt ctggattgcc tgacaatgcc agacgaggcg gtaccggcgg atccatggtg
1321 agtgcgatta agccagacat gaagatcaaa ctgcgtatgg aaggcaacgt aaacgggcac
1381 cactttgtga tcgacggaga tggtacaggc aagccttatg agggaaaaca gaccatggat
1441 cttgaagtca aagagggcgg acctctgcct tttgcctttg atatcctgac cactgcattc
1501 cattacggca acagggtatt cgtgaaatat ccagacaaca tacaagacta ttttaagcag
1561 tcgtttccta aggggtattc gtgggaacga agcatgactt tcgaagacgg gggcatttgc
1621 tatgccagaa gcgacataac aatggaaggg gacactttct ataataaagt tcgattttat
1681 ggtaccaact ttcccgccaa tggtccagtt atgcagaaga agacgctgaa atggatgccg
1741 agctggacgc gttcatcacg tcgtaagttc aataagacag gtaaggcact gagagctata
1801 ggtcggctga gctcaggagg ttctggaggt tctgattaca aggacgacga tgacaaggga
1861 ggttctggag gttcttaccc atacgacgta ccagattacg ctggaggttc tggaggttct
1921 gagcagaagc tgatctcaga ggaggacctg cgtacgtaaa agcttatcga taatcaacct
1981 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
2041 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
2101 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
2161 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
2221 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
2281 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
2341 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
2401 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
2461 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
2521 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga
2581 cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
2641 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
2701 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
2761 caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
2821 aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat
2881 cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
2941 gctcactgas kcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
3001 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
3061 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
3121 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
3181 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
3241 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
3301 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
3361 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
3421 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
3481 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
3541 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
3601 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
3661 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
3721 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
3781 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
3841 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
3901 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
3961 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
4021 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
4081 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
4141 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
4201 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
4261 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
4321 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
4381 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
4441 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
4501 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
4561 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
4621 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
4681 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
4741 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
4801 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
4861 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
4921 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
4981 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
5041 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
5101 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
5161 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
5221 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
5281 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
5341 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctgcg cgctcgctcg
5401 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
5461 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
5521 ttaatgatta acccgccatg ctacttatct acgtagccat gctctag
//
