| 质粒名称: | pAAV.Syn.Flex.GCaMP6s.WPRE.SV40 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 100845 |
| 存储实验室: | Douglas Kim, GENIE Project |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 5917 bp |
| 载体类型: | Mammalian Expression, AAV |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | GCaMP6s、GCaMP3 variant 643 |
| 物种来源: | Synthetic |
| 插入大小: | 1353 bp |
| 基因突变: | GCaMP3-K78H T302L R303P D380Y T381R S383T R392G |
| 启动子: | Synapsin |
| 克隆方法: | 未知 |
| 5’克隆位点: | 未知 |
| 3’克隆位点: | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: |
pAAV.Syn.Flex.GCaMP6s.WPRE.SV40 was a gift from Douglas Kim & GENIE Project (Addgene plasmid # 100845 ; http://n2t.net/addgene:100845 ; RRID:Addgene_100845)
For viral preps, please replace (Addgene plasmid # 100845) in the above sentence with: (Addgene viral prep # 100845-AAV1), (Addgene viral prep # 100845-AAV5), or (Addgene viral prep # 100845-AAV9)
|
| 参考文献部分: | Ultrasensitive fluorescent proteins for imaging neuronal activity. Chen TW, Wardill TJ, Sun Y, Pulver SR, Renninger SL, Baohan A, Schreiter ER, Kerr RA, Orger MB, Jayaraman V, Looger LL, Svoboda K, Kim DS. Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354. 10.1038/nature12354 PubMed 23868258 |
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.Syn.Flex.GCaMP6s.WPRE.SV40 5917 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION AAV expression of Cre recombinase-activated ultrasensitive protein
calcium sensor from Synapsin promoter.
ACCESSION .
VERSION .
KEYWORDS pAAV.Syn.Flex.GCaMP6s.WPRE.SV40
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5917)
AUTHORS Chen TW, Wardill TJ, Sun Y, Pulver SR, Renninger SL, Baohan A,
Schreiter ER, Kerr RA, Orger MB, Jayaraman V, Looger LL, Svoboda K,
Kim DS
TITLE Ultrasensitive fluorescent proteins for imaging neuronal activity.
JOURNAL Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354.
PUBMED 23868258
REFERENCE 2 (bases 1 to 5917)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5917
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 65..194
/label=AAV2 ITR
promoter 276..723
/label=hSyn promoter
/note="human synapsin I promoter; confers neuron-specific
expression (Kugler et al., 2003)"
protein_bind complement(753..786)
/label=lox2272
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GGATACTT). lox2272 sites are compatible with each
other, but incompatible with loxP or loxN sites (Lee and
Saito, 1988)."
protein_bind complement(849..882)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS complement(893..2245)
/codon_start=1
/product="improved fluorescent protein-based calcium sensor
(Chen et al., 2013)"
/label=GCaMP6s
/translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
primer_bind 1452..1471
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1711..1732
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1838..1859)
/label=EGFP-C
/note="EGFP, forward primer"
regulatory 2140..2149
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 2242..2251
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
protein_bind 2262..2295
/label=lox2272
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GGATACTT). lox2272 sites are compatible with each
other, but incompatible with loxP or loxN sites (Lee and
Saito, 1988)."
protein_bind 2366..2399
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 2441..3029
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2494..2514)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2912..2923)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(3032..3048)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3033..3049)
/label=pBluescriptKS
/note="For pBluescript vector"
polyA_signal 3078..3199
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3089..3108)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3143..3162
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(3534..3551)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3705..4293)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3785..3804)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4464..5324)
/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 5087..5106
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5325..5429)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5455..5910)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5592..5613)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5804..5823
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
61 ccagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac
121 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
181 cactaggggt tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg
241 ctctaggaag atctctgcag agggccctgc gtatgagtgc aagtgggttt taggaccagg
301 atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga caagcaccca
361 acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa caggatgcgg
421 cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc cccgcctggc
481 ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg
541 caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc agccggaccg
601 caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg cggcgccggc
661 gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg
721 cagtcgaatt caagctgcta gcaaggatcc ccataacttc gtataaagta tcctatacga
781 agttatatca aaataggaag accaatgctt caccatcgac ccgaattgcc aagcatcacc
841 atcgacccat aacttcgtat aatgtatgct atacgaagtt atactagagc tttcacttcg
901 ctgtcatcat ttgtacaaac tcttcgtagt ttacctgacc atccccatcg atgtctgctt
961 ccctgatcat ttcatcaacc tcttcatctg ttaacttctc tccaaggttt gtcatcacgt
1021 ggcgaagctc tgctgcactg atgtagccat tgccatcctt atcaaacaca ccgaacgctt
1081 ctctaatttc ttcttccgtg tccctgtatt tcatttttct tgccatcatt gtcaggaact
1141 cagggaagtc gattgtgccg tcaccgtcgg catctacttc attgatcatg tcctgcagct
1201 ctgcttctgt ggggttctgc cccagagacc gcatcaccgt ccccagctcc ttggttgtta
1261 ttgtcccatc cccgtccttg tcaaataggg agaaagcctc tttaaattct gcgatctgct
1321 cttcagtcag ttggtccggc aggttgtact ccagcttgtg ccccaggatg ttgccgtcct
1381 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
1441 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggatgt
1501 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc
1561 tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct
1621 tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcaccct
1681 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
1741 ccaccccggt gaacagctcc tcgcccttgc tcaccatgct ccctccggta ccgcccttgt
1801 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
1861 gatcgcgctt ctcgttgggg tctttcgaaa gtttggactg cacgctcagg tagtggttgt
1921 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg taggcgagct
1981 gcacgccgcc gtcctcgatg ttgtggcgga tgtggaagtt cgccttgatg ccgttcttct
2041 gcttgtcggc cttgatatag acgttctcga gtgagctcag ccgacctata gctctgactg
2101 cgtgacctgt cttattccac ttacgacgtg atgagtcgac catggtggcg agatccttat
2161 cgtcatcgtc gtacagatcc cgacccattt gctgtccacc agtcatgcta gccataccat
2221 gatgatgatg atgatgagaa cccatggtgg cggatcctag tataacttcg tataggatac
2281 tttatacgaa gttatcattg ggattcttcc tattttgatc caagcatcac catcgaccct
2341 ctagtccaga tctcaccatc gacccataac ttcgtatagc atacattata cgaagttatg
2401 tccctcgaag aggttcgaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
2461 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
2521 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
2581 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
2641 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
2701 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
2761 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
2821 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
2881 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
2941 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
3001 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgac ccgggcggcc
3061 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
3121 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
3181 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
3241 ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa
3301 ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca
3361 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
3421 ccgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcattaatga
3481 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
3541 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
3601 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
3661 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
3721 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
3781 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
3841 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
3901 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
3961 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
4021 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
4081 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
4141 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
4201 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
4261 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
4321 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
4381 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
4441 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
4501 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
4561 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
4621 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
4681 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
4741 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
4801 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
4861 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
4921 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
4981 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
5041 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
5101 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
5161 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
5221 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
5281 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
5341 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
5401 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
5461 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
5521 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
5581 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
5641 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
5701 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
5761 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
5821 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
5881 ccgcgcttaa tgcgccgcta cagggcgcgt cccattc
//
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