| 质粒名称: | pAAV-hSyn-iLMO (slGLuc-Mac-EYFP) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 114100 |
| 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: | 4563 bp |
| 完整质粒大小: | 7010 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, superluminescent,slGLuc |
| 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: | EcoRI (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-iLMO (slGLuc-Mac-EYFP) was a gift from Ute Hochgeschwender (Addgene plasmid # 114100 ; http://n2t.net/addgene:114100 ; RRID:Addgene_114100) |
| 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-iLMO (slGLuc-Mac-EYFP) 7086 bp ds-DNA circular SYN 12-AUG-2019
DEFINITION fusion protein of Gaussia luciferase variant superluminescent,
Leptosphaeria maculans opsin, and enhanced green fluorescent protein
for bioluminescent optogenetics.
ACCESSION .
VERSION .
KEYWORDS pAAV-hSyn-iLMO (slGLuc-Mac-EYFP)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7086)
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 7086)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 12, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7086
/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 652..661
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 1252..1261
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2452..3165
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(2494..2515)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2755..2774)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3102..3123
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 3217..3805
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3270..3290)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3688..3699)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3837..4313
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(4036..4055)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 4353..4493
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 4353..4482
/label=AAV2 ITR
rep_origin 4568..5023
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4655..4674)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4865..4886
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5040..5059)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5159..5181
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5219..5237)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5305..5409
/gene="bla"
/label=AmpR promoter
CDS 5410..6270
/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(5628..5647)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6441..7029
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6930..6949
/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 gatccaagct tgccaccatg
661 ggagtcaaag ttctgtttgc cctgatctgc atcgctgtgg ccgaggccaa gcccaccgag
721 aacaacgaag acttcaacat cgtggccgtg gccagcaact tcgcgaccac ggatctcgat
781 gctgaccgcg ggaagttgcc cggcaagaag ctgccgctgg aggtgctcaa agagatggaa
841 gccaatgccc ggaaagctgg ctgcaccagg ggctgtctga tctgcctgtc ccacatcaag
901 tgcacgccca agatgaagaa gtggctgcca ggacgctgcc acacctacga aggcgacaaa
961 gagtccgcac agggcggcat aggcgaggcg atcgtcgaca ttcctgagat tcctgggttc
1021 aaggacttgg agcccatgga gcagttcatc gcacaggtcg atctgtgtgt ggactgcaca
1081 actggctgcc tcaaagggct tgccaacgtg cagtgttctg acctgctcaa gaagtggctg
1141 ccgcaacgct gtgcgacctt tgccagcaag atccagggcc aggtggacaa gatcaagggg
1201 gccggtggtg acggcgaccc actagtccag tgtggtggaa ttgccggatc tgccaccatg
1261 gggacctgga tgctgacgaa ggctcgcgag gctgtgagca gccacagtgc cctgctcaga
1321 agccccaagc tcgtcaatca agctggttct ccatttgcac tcaggagcac gggcaggcga
1381 gtggccccta gttctggggg cagcggggga tccgccacca tgatcgtgga ccagttcgag
1441 gaggtgctga tgaagaccag ccagctgttc ccactgccaa ccgctaccca gagcgcccag
1501 ccaacccacg tggcccccgt gccaaccgtg ctgcccgaca cccccatcta cgagaccgtg
1561 ggcgacagcg gcagcaagac cctgtgggtg gtgttcgtgc tgatgctgat cgccagcgcc
1621 gccttcaccg ccctgagctg gaagatcccc gtgaacagga ggctgtacca cgtgatcacc
1681 accatcatca ccctgaccgc cgccctgagc tacttcgcta tggctaccgg ccacggagtg
1741 gccctgaaca agatcgtgat caggacccag cacgaccacg tgcccgacac ctacgagacc
1801 gtgtaccgac aggtgtacta cgccaggtac atcgactggg ctatcaccac cccactgctg
1861 ctgctggacc tgggactgct ggctggaatg agcggagccc acatcttcat ggccatcgtg
1921 gctgacctga tcatggtgct gaccggcctg ttcgctgctt tcggcagcga gggaacccca
1981 cagaagtggg gatggtacac catcgcctgc atcgcctaca tcttcgtggt gtggcacctg
2041 gtgctgaacg gcggcgccaa cgccagggtg aagggcgaga agctgaggag cttcttcgtg
2101 gccatcggag cttacaccct gatcctgtgg accgcttacc caatcgtgtg gggactggct
2161 gacggagcta ggaagatcgg agtggacgga gagatcatcg cttacgctgt gctggacgtg
2221 ctggctaagg gagtgttcgg agcttggctg ctggtgaccc acgccaacct gagggagagc
2281 gacgtggagc tgaacggctt ctgggccaac ggcctgaaca gggagggcgc catcaggatc
2341 ggcgaggacg acggcgcccg accggtagta gcagtgagca aggcggccgc caagagcagg
2401 atcaccagcg agggcgagta catccccctg gaccagatcg acatcaacgt ggtgagcaag
2461 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
2521 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
2581 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
2641 ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
2701 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
2761 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
2821 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
2881 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
2941 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
3001 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
3061 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
3121 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagttctg ctacgagaac
3181 gaggtgtaat gagaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
3241 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
3301 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
3361 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
3421 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
3481 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
3541 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
3601 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg
3661 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
3721 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
3781 atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt
3841 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
3901 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
3961 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
4021 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
4081 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
4141 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
4201 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
4261 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
4321 tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
4381 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
4441 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
4501 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
4561 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
4621 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
4681 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
4741 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
4801 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
4861 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
4921 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
4981 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
5041 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
5101 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
5161 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
5221 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
5281 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
5341 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
5401 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
5461 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
5521 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
5581 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
5641 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
5701 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
5761 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
5821 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
5881 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
5941 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
6001 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
6061 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
6121 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
6181 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
6241 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
6301 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
6361 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
6421 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
6481 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
6541 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
6601 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
6661 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
6721 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
6781 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
6841 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
6901 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
6961 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
7021 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
7081 gctcac
//
