| 质粒名称: | pAAV-Ef1a-DIO EYFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 27056 |
| 存储实验室: | Karl Deisseroth |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5603 bp |
| 载体类型: | 哺乳细胞表达,AAV |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | EYFP |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 720 bp |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AscI (未破坏) |
| 3' 克隆位点: | NheI (未破坏) |
| 5' 测序引物 | ggccagcttggcacttgatg |
| 3' 测序引物 | GCAATAGCATGATACAAAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-Ef1a-DIO EYFP was a gift from Karl Deisseroth (Addgene plasmid # 27056 ; http://n2t.net/addgene:27056 ; RRID:Addgene_27056) |
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-Ef1a-DIO EYFP 6313 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pAAV-Ef1a-DIO EYFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6313)
TITLE Double Floxed Inverted ORF Control
REFERENCE 2 (bases 1 to 6313)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6313
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..130
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
promoter 231..1409
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 462..1400
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1357..1377
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
protein_bind 1440..1473
/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."
protein_bind 1524..1557
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS complement(1566..2285)
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1611..1632)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 1960..1979
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2219..2240
/label=EGFP-N
/note="EGFP, reverse primer"
regulatory 2282..2291
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
protein_bind complement(2298..2331)
/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."
protein_bind complement(2382..2415)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 2440..3028
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2493..2513)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2911..2922)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3060..3536
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3259..3278)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 3576..3716
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3576..3705
/label=AAV2 ITR
rep_origin 3791..4246
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3878..3897)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4088..4109
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4263..4282)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4382..4404
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4442..4460)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4528..4632
/gene="bla"
/label=AmpR promoter
CDS 4633..5493
/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(4851..4870)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5664..6252
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6153..6172
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
1561 gcgccttact tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
1621 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctggtagctc
1681 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
1741 tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg
1801 atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc
1861 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
1921 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
1981 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
2041 tgctgcttca tgtggtcggg gtagcgggcg aagcactgca ggccgtagcc gaaggtggtc
2101 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc
2161 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
2221 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
2281 accatggtgg cgctagcata acttcgtata aagtatccta tacgaagtta tttgccttaa
2341 cccagaaatt atcactgtta ttctttagaa tggtgcaaag aataacttcg tataatgtat
2401 gctatacgaa gttatgaatt cgatatcaag cttatcgata atcaacctct ggattacaaa
2461 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
2521 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
2581 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
2641 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
2701 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
2761 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
2821 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt
2881 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
2941 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
3001 cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg
3061 ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag
3121 tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct
3181 tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa
3241 cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc
3301 tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt
3361 tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg
3421 gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt
3481 ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga
3541 ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
3601 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
3661 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
3721 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
3781 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
3841 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
3901 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
3961 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
4021 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
4081 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
4141 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
4201 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
4261 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
4321 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
4381 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
4441 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
4501 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
4561 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
4621 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
4681 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
4741 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
4801 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
4861 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
4921 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
4981 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
5041 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
5101 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
5161 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
5221 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
5281 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
5341 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
5401 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
5461 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
5521 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
5581 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
5641 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
5701 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
5761 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
5821 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
5881 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
5941 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
6001 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
6061 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
6121 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
6181 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
6241 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
6301 tttgctcaca tgt
//
