| 质粒名称: | pAAV-hSyn-LMO3 (sbGLuc-VChR1-EYFP) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 114099 |
| 存储实验室: | Ute Hochgeschwender |
|
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: | 4604 bp |
| 完整质粒大小: | 6952 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,AAV |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Gaussia luciferase, slow burn,sbGLuc |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | hSyn |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | SacI(未破坏) |
| 3' 克隆位点: | NheI(未破坏) |
| 5' 测序引物 | ACTGAAGGCGCGCTGACGTCACT |
| 3' 测序引物 | CATAGCGTAAAAGGAGCAACA |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-hSyn-LMO3 (sbGLuc-VChR1-EYFP) was a gift from Ute Hochgeschwender (Addgene plasmid # 114099 ; http://n2t.net/addgene:114099 ; RRID:Addgene_114099) |
| 参考文献部分: | Luminopsins integrate opto- and chemogenetics by using physical and biological light sources for opsin activation. Berglund K, Clissold K, Li HE, Wen L, Park SY, Gleixner J, Klein ME, Lu D, Barter JW, Rossi MA, Augustine GJ, Yin HH, Hochgeschwender U. Proc Natl Acad Sci U S A. 2016 Jan 19;113(3):E358-67. doi: 10.1073/pnas.1510899113. Epub 2016 Jan 5. 10.1073/pnas.1510899113 PubMed 26733686 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pAAV-hSyn-LMO3 (sbGLuc-VChR1-EYFP) 6937 bp ds-DNA circular SYN 06-AUG-2019
DEFINITION fusion protein of Gaussia luciferase variant slow burn, Volvox
channelrhodopsin-1, and enhanced yellow fluorescent protein for
bioluminescent optogenetics.
ACCESSION .
VERSION .
KEYWORDS pAAV-hSyn-LMO3 (sbGLuc-VChR1-EYFP)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6937)
AUTHORS Berglund K, Clissold K, Li HE, Wen L, Park SY, Gleixner J, Klein ME,
Lu D, Barter JW, Rossi MA, Augustine GJ, Yin HH, Hochgeschwender U
TITLE Luminopsins integrate opto- and chemogenetics by using physical and
biological light sources for opsin activation.
JOURNAL Proc Natl Acad Sci U S A. 2016 Jan 19;113(3):E358-67. doi:
10.1073/pnas.1510899113. Epub 2016 Jan 5.
PUBMED 26733686
REFERENCE 2 (bases 1 to 6937)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6937
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 5..134
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
promoter 179..626
/label=hSyn promoter
/note="human synapsin I promoter; confers neuron-specific
expression (Kugler et al., 2003)"
regulatory 687..696
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 693..1247
/codon_start=1
/product="enhanced variant of secreted Gaussia luciferase"
/label=Gaussia-Dura Luc
/note="human codon-optimized"
/translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
RCATFASKIQGQVDKIKGAGGD"
CDS 693..1247
/codon_start=1
/product="secreted Gaussia luciferase"
/label=hGLuc
/note="human codon-optimized"
/translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
RCATFASKIQGQVDKIKGAGGD"
regulatory 1287..1296
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2262..2975
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(2304..2325)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2565..2584)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2912..2933
/label=EGFP-C
/note="EGFP, forward primer"
CDS 3020..3037
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
misc_feature 3068..3656
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3121..3141)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3539..3550)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3688..4164
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3887..3906)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 4204..4344
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 4204..4333
/label=AAV2 ITR
rep_origin 4419..4874
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4506..4525)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4716..4737
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4891..4910)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5010..5032
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5070..5088)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5156..5260
/gene="bla"
/label=AmpR promoter
CDS 5261..6121
/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(5479..5498)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6292..6880
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6781..6800
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac
61 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
121 cactaggggt tcctgcggcc gcacgcgtgt gtctagactg cagagggccc tgcgtatgag
181 tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc
241 ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga
301 gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga
361 cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc
421 tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg
481 ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg
541 accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag
601 gagtcgtgtc gtgcctgaga gcgcagtcga gaaggtaccg gatcctctag agtcgagctc
661 accggtggcg cgccaccggt aagcttgcca ccatgggagt caaagttctg tttgccctga
721 tctgcatcgc tgtggccgag gccaagccca ccgagaacaa cgaagacttc aacatcgtgg
781 ccgtggccag caacttcgcg accacggatc tcgatgctga ccgcgggaag ttgcccggca
841 agaagctgcc gctggaggtg ctcaaagagc tggaagccaa tgcccggaaa gctggctgca
901 ccaggggctg tctgatctgc ctgtcccaca tcaagtgcac gcccaagatg aagaagttca
961 tcccaggacg ctgccacacc tacgaaggcg acaaagagtc cgcacagggc ggcataggcg
1021 aggcgatcgt cgacattcct gagattcctg ggttcaagga cttggagccc ctggagcagt
1081 tcatcgcaca ggtcgatctg tgtgtggact gcacaactgg ctgcctcaaa gggcttgcca
1141 acgtgcagtg ttctgacctg ctcaagaagt ggctgccgca acgctgtgcg acctttgcca
1201 gcaagatcca gggccaggtg gacaagatca agggggccgg tggtgacggc gacccactag
1261 tccagtgtgg tggaattgcc ggatccgcca ccatggatta ccctgttgct agaagcctca
1321 tagttcgcta cccaaccgac ctcggaaacg gcaccgtctg catgccaaga ggacagtgtt
1381 actgtgaagg ttggcttcgg agtcgcggca cttccattga aaagacaata gcaattactc
1441 ttcagtgggt agtctttgct ttgtcagtgg cttgcctggg gtggtatgcg tatcaagcgt
1501 ggcgagctac ctgcggatgg gaggaggttt acgtagcctt gatagaaatg atgaaaagca
1561 tcatcgaggc cttccacgag ttcgacagcc ctgcaacact gtggctgtct tcagggaacg
1621 gcgtagtttg gatgcggtat ggcgaatggc tcctcacctg cccggtcctt ctgatccatc
1681 tgagcaacct cacaggcctg aaggacgatt atagcaaaag gactatgggc ctgttggttt
1741 ctgatgtggg atgcatcgtg tggggcgcaa ccagcgccat gtgtacgggg tggacgaaga
1801 tcctgttctt cctcatctca ttgagctatg gtatgtatac ctattttcat gctgctaaag
1861 tttatatcga agcattccac acagttccaa aagggatttg tcgagaactg gtccgagtga
1921 tggcctggac attctttgtg gcttggggaa tgtttccagt cctgtttctg ctgggcacgg
1981 aaggattcgg tcatatcagc ccttatggat ctgccattgg gcactccatc ctcgacctga
2041 ttgcaaagaa catgtggggt gtgctgggga attacctgcg cgtcaaaatc cacgagcaca
2101 tcctgttgta tggcgacatc agaaagaagc agaaaattac gatcgccggc caagagatgg
2161 aggttgagac actggtggct gaagaggagg acgcggccgc caagagcagg atcaccagcg
2221 agggcgagta catccccctg gaccagatcg acatcaacgt ggtgagcaag ggcgaggagc
2281 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
2341 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
2401 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg
2461 gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
2521 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
2581 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
2641 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
2701 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
2761 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
2821 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc
2881 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
2941 ccgggatcac tctcggcatg gacgagctgt acaagttctg ctacgagaac gaggtgtaat
3001 gagctagcca attgccggtc atcatcacca tcaccattga gtttaattcg atatcaagct
3061 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
3121 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
3181 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
3241 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
3301 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
3361 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
3421 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
3481 gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
3541 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
3601 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
3661 ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct
3721 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
3781 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
3841 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
3901 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
3961 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
4021 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
4081 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
4141 accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc
4201 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
4261 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
4321 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
4381 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
4441 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
4501 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
4561 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
4621 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
4681 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
4741 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
4801 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
4861 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
4921 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
4981 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
5041 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
5101 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
5161 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
5221 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
5281 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
5341 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
5401 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
5461 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
5521 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
5581 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
5641 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
5701 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
5761 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
5821 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
5881 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
5941 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
6001 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
6061 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
6121 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
6181 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
6241 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
6301 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
6361 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
6421 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
6481 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
6541 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
6601 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
6661 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
6721 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
6781 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
6841 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
6901 tttacggttc ctggcctttt gctggccttt tgctcac
//
