| 质粒名称: | pAAP2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 73211 |
| 存储实验室: | Luk Vandenberghe |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达,AAV |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | AAP2 |
| 物种来源: | AAV2 |
| 插入大小: | 612 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | T7 |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pAAP2 was a gift from Luk Vandenberghe (Addgene plasmid # 73211 ; http://n2t.net/addgene:73211 ; RRID:Addgene_73211) |
| 参考文献部分: | In Silico Reconstruction of the Viral Evolutionary Lineage Yields a Potent Gene Therapy Vector. Zinn E, Pacouret S, Khaychuk V, Turunen HT, Carvalho LS, Andres-Mateos E, Shah S, Shelke R, Maurer AC, Plovie E, Xiao R, Vandenberghe LH. Cell Rep. 2015 Jul 28. pii: S2211-1247(15)00759-7. doi: 10.1016/j.celrep.2015.07.019. 10.1016/j.celrep.2015.07.019 PubMed 26235624 |
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 pAAP2 5992 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION express Assembly-activating protein (AAP) of adeno-associated virus
serotype 2 (AAV2).
ACCESSION .
VERSION .
KEYWORDS pAAP2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5992)
AUTHORS Zinn E, Pacouret S, Khaychuk V, Turunen HT, Carvalho LS,
Andres-Mateos E, Shah S, Shelke R, Maurer AC, Plovie E, Xiao R,
Vandenberghe LH
TITLE In Silico Reconstruction of the Viral Evolutionary Lineage Yields a
Potent Gene Therapy Vector.
JOURNAL Cell Rep. 2015 Jul 28. pii: S2211-1247(15)00759-7. doi:
10.1016/j.celrep.2015.07.019.
PUBMED 26235624
REFERENCE 2 (bases 1 to 5992)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5992
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1586..1603)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1592..1816
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1862..2290
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1949..1968)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2159..2180
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2299..2319)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2304..2633
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2484..2619
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2546..2565
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2700..3494
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2754..2773)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3364..3383
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3668..3789
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3705..3724)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3759..3778
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3838..3854)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3838..3854)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3851..3873)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3862..3878
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(3886..3916)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3931..3952
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4069..4086)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4240..4825)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4320..4339)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4996..5856)
/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 5619..5638
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5857..5961)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 gtttaaacgg gccctctaga ctcgagatgg agacgcagac tcagtacctg acccccagcc
961 tctcggacag ccaccagcag ccccctctgg tctgggaact aatacgatgg ctacaggcag
1021 tggcgcacca atggcagaca ataacgaggg cgccgacgga gtgggtaatt cctcgggaaa
1081 ttggcattgc gattccacat ggatgggcga cagagtcatc accaccagca cccgaacctg
1141 ggccctgccc acctacaaca accacctcta caaacaaatt tccagccaat caggagcctc
1201 gaacgacaat cactactttg gctacagcac cccttggggg tattttgact tcaacagatt
1261 ccactgccac ttttcaccac gtgactggca aagactcatc aacaacaact ggggattccg
1321 acccaagaga ctcaacttca agctctttaa cattcaagtc aaagaggtca cgcagaatga
1381 cggtacgacg acgattgcca ataaccttac cagcacggtt caggtgttta ctgactcgga
1441 gtaccagctc ccgtacgtcc tcggctcggc gcatcaagga tgcctcccgc cgttcccagc
1501 agacgtcttc atggtgccac agtatggata cctcaccctg aggatccgag ctcggtacca
1561 agcttaagtt taaaccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
1621 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
1681 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
1741 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
1801 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
1861 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
1921 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
1981 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
2041 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
2101 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
2161 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
2221 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
2281 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
2341 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
2401 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
2461 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
2521 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
2581 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
2641 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
2701 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
2761 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
2821 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
2881 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
2941 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
3001 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
3061 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
3121 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
3181 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
3241 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
3301 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
3361 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
3421 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
3481 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
3541 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
3601 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
3661 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
3721 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
3781 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
3841 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
3901 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
3961 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
4021 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
4081 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
4141 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
4201 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
4261 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
4321 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
4381 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
4441 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
4501 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
4561 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
4621 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
4681 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
4741 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
4801 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
4861 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
4921 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
4981 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
5041 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
5101 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
5161 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
5221 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
5281 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
5341 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
5401 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
5461 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
5521 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
5581 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
5641 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
5701 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
5761 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
5821 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
5881 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
5941 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
