| 质粒名称: | pAAV-hSyn-EGFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 50465 |
| 存储实验室: | Bryan Roth |
|
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: | 4818 bp |
| 完整质粒大小: | 5265 bp |
| 载体修饰: | -- |
| 载体类型: | AAV |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | EGFP |
| 物种来源: | 维多利亚多管发光水母 |
| 插入大小: | 726 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | human Synapsin 1 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamH I(未破坏) |
| 3' 克隆位点: | EcoR I (未破坏) |
| 5' 测序引物 | TCGTGTCGTGCCTGAGAGCG |
| 3' 测序引物 | GCATTAAAGCAGCGTATCCACATAGC |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-hSyn-EGFP was a gift from Bryan Roth (Addgene plasmid # 50465 ; http://n2t.net/addgene:50465 ; RRID:Addgene_50465) |
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-EGFP 5265 bp ds-DNA circular SYN 09-APR-2019
DEFINITION EGFP under the control of human synapsin promoter.
ACCESSION .
VERSION .
KEYWORDS pAAV-hSyn-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5265)
AUTHORS Bryan Roth
TITLE Roth lab DREADDs
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5265)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 9, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5265
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 6..135
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
promoter 180..627
/label=hSyn promoter
/note="human synapsin I promoter; confers neuron-specific
expression (Kugler et al., 2003)"
regulatory 647..656
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 653..1372
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(698..719)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(959..978)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1306..1327
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 1397..1985
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(1450..1470)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(1868..1879)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 2017..2493
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(2216..2235)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 2533..2673
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 2533..2662
/label=AAV2 ITR
rep_origin 2748..3203
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2835..2854)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3045..3066
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3220..3239)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3339..3361
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3399..3417)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3485..3589
/gene="bla"
/label=AmpR promoter
CDS 3590..4450
/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(3808..3827)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4621..5209
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5110..5129
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
121 tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc ctgcgtatga
181 gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac
241 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
301 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
361 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
421 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
481 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
541 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
601 ggagtcgtgt cgtgcctgag agcgcagtcg agaaggtacc ggatccgcca ccatggtgag
661 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
721 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
781 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
841 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
901 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
961 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
1021 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
1081 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
1141 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
1201 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
1261 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
1321 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aggaattcga
1381 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
1441 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
1501 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
1561 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
1621 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
1681 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
1741 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
1801 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta
1861 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
1921 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
1981 cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc
2041 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
2101 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
2161 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
2221 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
2281 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc
2341 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
2401 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
2461 caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga
2521 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
2581 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
2641 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
2701 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
2761 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
2821 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
2881 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
2941 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
3001 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
3061 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
3121 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
3181 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
3241 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
3301 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
3361 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
3421 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
3481 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
3541 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
3601 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
3661 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
3721 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
3781 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
3841 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
3901 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
3961 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
4021 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
4081 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
4141 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
4201 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
4261 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
4321 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
4381 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
4441 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
4501 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
4561 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
4621 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
4681 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
4741 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
4801 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
4861 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
4921 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
4981 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
5041 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
5101 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
5161 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
5221 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctca
//
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