| 质粒名称: | 7M8 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 64839 |
| 存储实验室: | John Flannery, David Schaffer |
|
载体骨架基本信息 |
|
| 载体名称: | XX2 |
| 载体出品公司: | University of North Carolina Dr. Samulski |
| 空载体大小: | 5648 bp |
| 完整质粒大小: | 8209 bp |
| 载体类型: | AAV |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | 7M8 cap |
| 物种来源: | Adeno-associated virus - 2 |
| 插入大小: | 2238 bp |
| 基因突变: | 7mer insertion in the AAV2 cap |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | HindIII (未破坏) |
| 3’克隆位点: | NotI (未破坏) |
| 5’测序引物 | GCGGAAGCTTCGATCAACTACGC |
| 3’测序引物 | gtgcggcaaagtttgcttcc |
|
文献引用方法 |
|
| 材料和方法部分: | 7M8 was a gift from John Flannery & David Schaffer (Addgene plasmid # 64839 ; http://n2t.net/addgene:64839 ; RRID:Addgene_64839) |
| 参考文献部分: | In vivo-directed evolution of a new adeno-associated virus for therapeutic outer retinal gene delivery from the vitreous. Dalkara D, Byrne LC, Klimczak RR, Visel M, Yin L, Merigan WH, Flannery JG, Schaffer DV. Sci Transl Med. 2013 Jun 12;5(189):189ra76. doi: 10.1126/scitranslmed.3005708. 10.1126/scitranslmed.3005708 PubMed 23761039 |
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 7M8 8369 bp ds-DNA circular SYN 09-SEP-2021
DEFINITION This plasmid can be used in the triple transfection method of AAV
vector production. It supplies the replication proteins from AAV2
and the 7M8 capsid protein..
ACCESSION .
VERSION .
KEYWORDS 7M8
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8369)
AUTHORS Dalkara D, Byrne LC, Klimczak RR, Visel M, Yin L, Merigan WH,
Flannery JG, Schaffer DV
TITLE In vivo-directed evolution of a new adeno-associated virus for
therapeutic outer retinal gene delivery from the vitreous.
JOURNAL Sci Transl Med. 2013 Jun 12;5(189):189ra76. doi:
10.1126/scitranslmed.3005708.
PUBMED 23761039
REFERENCE 2 (bases 1 to 8369)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 9, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8369
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 8..110
/label=ITR
/note="inverted terminal repeat of human adenovirus
serotype 5"
CDS 4213..4224
/codon_start=1
/product="WELQut protease recognition and cleavage site"
/label=WELQut site
/translation="WELQ"
rep_origin 5150..5663
/direction=RIGHT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5237..5256)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5447..5468
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6145..6164)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6264..6286
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6324..6342)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6410..6514
/gene="bla"
/label=AmpR promoter
CDS 6515..7375
/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(6733..6752)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7546..8134
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8035..8054
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8288..8305
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 gaattcccat catcaataat ataccttatt ttggattgaa gccaatatga taatgagggg
61 gtggagtttg tgacgtggcg cggggcgtgg gaacggggcg ggtgacgtag tagctctaga
121 ggtcctgtat tagaggtcac gtgagtgttt tgcgacattt tgcgacacca tgtggtcacg
181 ctgggtattt aagcccgagt gagcacgcag ggtctccatt ttgaagcggg aggtttgaac
241 gcgcagccac cacggcgggg ttttacgaga ttgtgattaa ggtccccagc gaccttgacg
301 agcatctgcc cggcatttct gacagctttg tgaactgggt ggccgagaag gaatgggagt
361 tgccgccaga ttctgacatg gatctgaatc tgattgagca ggcacccctg accgtggccg
421 agaagctgca gcgcgacttt ctgacggaat ggcgccgtgt gagtaaggcc ccggaggccc
481 ttttctttgt gcaatttgag aagggagaga gctacttcca catgcacgtg ctcgtggaaa
541 ccaccggggt gaaatccatg gttttgggac gtttcctgag tcagattcgc gaaaaactga
601 ttcagagaat ttaccgcggg atcgagccga ctttgccaaa ctggttcgcg gtcacaaaga
661 ccagaaatgg cgccggaggc gggaacaagg tggtggatga gtgctacatc cccaattact
721 tgctccccaa aacccagcct gagctccagt gggcgtggac taatatggaa cagtatttaa
781 gcgcctgttt gaatctcacg gagcgtaaac ggttggtggc gcagcatctg acgcacgtgt
841 cgcagacgca ggagcagaac aaagagaatc agaatcccaa ttctgatgcg ccggtgatca
901 gatcaaaaac ttcagccagg tacatggagc tggtcgggtg gctcgtggac aaggggatta
961 cctcggagaa gcagtggatc caggaggacc aggcctcata catctccttc aatgcggcct
1021 ccaactcgcg gtcccaaatc aaggctgcct tggacaatgc gggaaagatt atgagcctga
1081 ctaaaaccgc ccccgactac ctggtgggcc agcagcccgt ggaggacatt tccagcaatc
1141 ggatttataa aattttggaa ctaaacgggt acgatcccca atatgcggct tccgtctttc
1201 tgggatgggc cacgaaaaag ttcggcaaga ggaacaccat ctggctgttt gggcctgcaa
1261 ctaccgggaa gaccaacatc gcggaggcca tagcccacac tgtgcccttc tacgggtgcg
1321 taaactggac caatgagaac tttcccttca acgactgtgt cgacaagatg gtgatctggt
1381 gggaggaggg gaagatgacc gccaaggtcg tggagtcggc caaagccatt ctcggaggaa
1441 gcaaggtgcg cgtggaccag aaatgcaagt cctcggccca gatagacccg actcccgtga
1501 tcgtcacctc caacaccaac atgtgcgccg tgattgacgg gaactcaacg accttcgaac
1561 accagcagcc gttgcaagac cggatgttca aatttgaact cacccgccgt ctggatcatg
1621 actttgggaa ggtcaccaag caggaagtca aagacttttt ccggtgggca aaggatcacg
1681 tggttgaggt ggagcatgaa ttctacgtca aaaagggtgg agccaagaaa agacccgccc
1741 ccagtgacgc agatataagt gagcccaaac gggtgcgcga gtcagttgcg cagccatcga
1801 cgtcagacgc ggaagcttcg atcaactacg cagacaggta ccaaaacaaa tgttctcgtc
1861 acgtgggcat gaatctgatg ctgtttccct gcagacaatg cgtgagaatg aatcagaatt
1921 caaatatctg cttcactcac ggacagaaag actgtttaga gtgctttccc gtgtcagaat
1981 ctcaacccgt ttctgtcgtc aaaaaggcgt atcagaaact gtgctacatt catcatatca
2041 tgggaaaggt gccagacgct tgcactgcct gcgatctggt caatgtggat ttggatgact
2101 gcatctttga acaataaatg atttaaatca ggtatggctg ccgatggtta tcttccagat
2161 tggctcgagg acactctctc tgaaggaata agacagtggt ggaagctcaa acctggccca
2221 ccaccaccaa agcccgcaga gcggcataag gacgacagca ggggtcttgt gcttcctggg
2281 tacaagtacc tcggaccctt caacggactc gacaagggag agccggtcaa cgaggcagac
2341 gccgcggccc tcgagcacga caaagcctat gaccggcagc tcgacagcgg agacaacccg
2401 tacctcaagt acaaccacgc cgacgcggag tttcaggagc gccttaaaga agatacgtct
2461 tttgggggca acctcggacg agcagtcttc caggcgaaaa agagggttct tgaacctctg
2521 ggcctggttg aggaacctgt taagacggct ccgggaaaaa agaggccggt agagcactct
2581 cctgtggagc cagactcctc ctcgggaacc ggaaaggcgg gccagcagcc tgcaagaaaa
2641 agattgaatt ttggtcagac tggagacgca gactcagtac ctgaccccca gcctctcgga
2701 cagccaccag cagccccctc tggtctggga actaatacga tggctacagg cagtggcgca
2761 ccaatggcag acaataacga gggcgccgac ggagtgggta attcctcggg aaattggcat
2821 tgcgattcca catggatggg cgacagagtc accaccacca gcacccgaac ctgggccctg
2881 cccacctaca acaaccacct ctacaaacaa atttccagcc aatcaggagc ctcgaacgac
2941 aatcactact ttggctacag caccccttgg gggtattttg acttcaacag attccactgc
3001 cacttttcac cacgtgactg gcaaagactc atcaacaaca actggggatt ccgacccaag
3061 agactcaact tcaagctctt taacattcaa gtcaaagagg tcacgcagaa tgacggtacg
3121 acgacgattg ccaataacct taccagcacg gttcaggtgt ttactgactc ggagtaccag
3181 ctcccgtacg tcctcggctc ggcgcatcaa ggatgcctcc cgccgttccc agcagacgtc
3241 ttcatggtgc cacagtatgg atacctcacc ctgaacaacg ggagtcaggc agtaggacgc
3301 tcttcatttt actgcctgga gtactttcct tctcagatgc tgcgtaccgg aaacaacttt
3361 accttcagct acacttttga ggacgttcct ttccacagca gctacgctca cagccagagt
3421 ctggaccgtc tcatgaatcc tctcatcgac cagtacctgt attacttgag cagaacaaac
3481 actccaagtg gaaccaccac gcagtcaagg cttcagtttt ctcaggccgg agcgagtgac
3541 attcgggacc agtctaggaa ctggcttcct ggaccctgtt accgccagca gcgagtatca
3601 aagacatctg cggataacaa caacagtgaa tactcgtgga ctggagctac caagtaccac
3661 ctcaatggca gagactctct ggtgaatccg ggcccggcca tggcaagcca caaggacgat
3721 gaagaaaagt tttttcctca gagcggggtt ctcatctttg ggaagcaagg ctcagagaaa
3781 acaaatgtgg acattgaaaa ggtcatgatt acagacgaag aggaaatcag gacaaccaat
3841 cccgtggcta cggagcagta tggttctgta tctaccaacc tccagagagg caacctagca
3901 ctcggcgaaa caacaagacc tgctaggcaa gcagctaccg cagatgtcaa cacacaaggc
3961 gttcttccag gcatggtctg gcaggacaga gatgtgtacc ttcaggggcc catctgggca
4021 aagattccac acacggacgg acattttcac ccctctcccc tcatgggtgg attcggactt
4081 aaacaccctc ctccccagat tctcatcaag aacaccccgg tacctgcgaa tccttcgacc
4141 accttcagtg cggcaaagtt tgcttccttc atcacacagt actccacggg acaggtcagc
4201 gtggagatcg agtgggagct gcagaaggaa aacagcaaac gctggaatcc cgaaattcag
4261 tacacttcca actacaacaa gtctattaat gtggacttta ctgtggacac taatggcgtg
4321 tattcagagc ctcgccccat tggcaccaga tacctgactc gtaatctgta attgcggccg
4381 cttgttaatc aataaaccgt ttaattcgtt tcagttgaac tttggtctct gcgtatttct
4441 ttcttatcta gtttccatgc tctagaggtc ctgtattaga ggtcacgtga gtgttttgcg
4501 acattttgcg acaccatgtg gtcacgctgg gtatttaagc ccgagtgagc acgcagggtc
4561 tccattttga agcgggaggt ttgaacgcgc agccaccacg gcggggtttt acgagattgt
4621 gattaaggtc cccagcgacc ttgacgagca tctgcccggc atttctgaca gctttgtgaa
4681 ctgggtggcc gagaaggaat gggagttgcc gccagattct gacatggatc tgaatctgat
4741 tgagcaggca cccctgaccg tggccgagaa gctgcatcgc tggcgtaata gcgaagaggc
4801 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gattccgttg
4861 caatggctgg cggtaatatt gttctggata ttaccagcaa ggccgatagt ttgagttctt
4921 ctactcaggc aagtgatgtt attactaatc aaagaagtat tgcgacaacg gttaatttgc
4981 gtgatggaca gactctttta ctcggtggcc tcactgatta taaaaacact tctcaggatt
5041 ctggcgtacc gttcctgtct aaaatccctt taatcggcct cctgtttagc tcccgctctg
5101 attctaacga ggaaagcacg ttatacgtgc tcgtcaaagc aaccatagta cgcgccctgt
5161 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
5221 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
5281 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
5341 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
5401 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
5461 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
5521 ccgatttcgg catattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
5581 aacaaaatat taacgcttac aatttaaata tttgcttata caatcttcct gtttttgggg
5641 cttttctgat tatcaaccgg ggtacatatg attgacatgc tagttttacg attaccgttc
5701 atcgattctc ttgtttgctc cagactctca ggcaatgacc tgatagcctt tgtagagacc
5761 tctcaaaaat agctaccctc tccggcatga atttatcagc tagaacggtt gaatatcata
5821 ttgatggtga tttgactgtc tccggccttt ctcacccgtt tgaatcttta cctacacatt
5881 actcaggcat tgcatttaaa atatatgagg gttctaaaaa tttttatcct tgcgttgaaa
5941 taaaggcttc tcccgcaaaa gtattacagg gtcataatgt ttttggtaca accgatttag
6001 ctttatgctc tgaggcttta ttgcttaatt ttgctaattc tttgccttgc ctgtatgatt
6061 tattggatgt tggaatcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
6121 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
6181 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
6241 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
6301 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
6361 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
6421 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
6481 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
6541 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
6601 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
6661 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
6721 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
6781 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
6841 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
6901 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
6961 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
7021 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
7081 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
7141 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
7201 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
7261 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
7321 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
7381 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
7441 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
7501 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
7561 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
7621 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
7681 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
7741 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
7801 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
7861 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
7921 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
7981 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
8041 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
8101 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
8161 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
8221 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
8281 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
8341 ccccgcgcgt tggccgattc attaatgca
//
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