| 质粒名称: | pZT-AAVS1-R1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 52638 |
| 存储实验室: | Mahendra Rao, Jizhong Zou |
|
载体骨架基本信息 |
|
| 载体名称: | pZT |
| 载体修饰: | Compatible with Golden Gate TALEN kit |
| 载体类型: | 哺乳细胞表达,TALEN |
| 载体抗性: | 卡那霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Right AAVS1 TALEN |
| 物种来源: | Synthetic |
| 插入大小: | 3009 bp |
| 启动子: | CMV |
| 5’测序引物 | TCGCGAAGAGAGGGGGAGTAAC |
| 3’测序引物 | ACCAAGACATGCCAACGCCACC |
|
文献引用方法 |
|
| 材料和方法部分: | pZT-AAVS1-R1 was a gift from Mahendra Rao & Jizhong Zou (Addgene plasmid # 52638 ; http://n2t.net/addgene:52638 ; RRID:Addgene_52638) |
| 参考文献部分: | Stable Enhanced Green Fluorescent Protein Expression After Differentiation and Transplantation of Reporter Human Induced Pluripotent Stem Cells Generated by AAVS1 Transcription Activator-Like Effector Nucleases. Luo Y, Liu C, Cerbini T, San H, Lin Y, Chen G, Rao MS, Zou J. Stem Cells Transl Med. 2014 May 15. pii: sctm.2013-0212. 10.5966/sctm.2013-0212 PubMed 24833591 |
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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 pZT-AAVS1-R1 5973 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION TALEN-mediated gene editing at human PPP1R12C/AAVS1 locus .
ACCESSION .
VERSION .
KEYWORDS pZT-AAVS1-R1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5973)
AUTHORS Luo Y, Liu C, Cerbini T, San H, Lin Y, Chen G, Rao MS, Zou J
TITLE Stable Enhanced Green Fluorescent Protein Expression After
Differentiation and Transplantation of Reporter Human Induced
Pluripotent Stem Cells Generated by AAVS1 Transcription
Activator-Like Effector Nucleases.
JOURNAL Stem Cells Transl Med. 2014 May 15. pii: sctm.2013-0212.
PUBMED 24833591
REFERENCE 2 (bases 1 to 5973)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5973
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 36..415
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 416..619
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 570..590
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 664..683
/label=T7
/note="T7 promoter, forward primer"
promoter 664..682
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 751..816
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 823..843
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3160..3747
/codon_start=1
/product="nonspecific DNA cleavage domain of the FokI
endonuclease (Li et al., 1992)"
/label=FokI cleavage domain
/note="must dimerize to be catalytically active"
/translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
primer_bind complement(3797..3814)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3803..4027
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
CDS 4200..4994
/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
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4254..4273)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4864..4883
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
rep_origin 5320..5908
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5809..5828
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gactcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
61 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
121 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
181 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
241 ctatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
301 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
361 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
421 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
481 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
541 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
601 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
661 aattaatacg actcactata gggagaccca agctggctag cgtttaaact taagctgatc
721 cactagtcca gtgtggtgga attcgccatg gactacaaag accatgacgg tgattataaa
781 gatcatgaca tcgattacaa ggatgacgat gacaagatgg cccccaagaa gaagaggaag
841 gtgggcatcc acggggtacc tatggtggac ttgaggacac tcggttattc gcaacagcaa
901 caggagaaaa tcaagcctaa ggtcaggagc accgtcgcgc aacaccacga ggcgcttgtg
961 gggcatggct tcactcatgc gcatattgtc gcgctttcac agcaccctgc ggcgcttggg
1021 acggtggctg tcaaatacca agatatgatt gcggccctgc ccgaagccac gcacgaggca
1081 attgtagggg tcggtaaaca gtggtcggga gcgcgagcac ttgaggcctt gctgactgtg
1141 gcgggtgagc ttagggggcc tccgctccag ctcgacaccg ggcagctgct gaagatcgcg
1201 aagagagggg gagtaacagc ggtagaggca gtgcacgcct ggcgcaatgc gctcaccggg
1261 gcccccctga acctgacccc ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag
1321 caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc
1381 ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg
1441 cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct
1501 atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg
1561 ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc
1621 ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc
1681 ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa
1741 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg
1801 gtggctatcg ccagcaacaa tggcggcaag caagcgctcg aaacggtgca gcggctgttg
1861 ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac
1921 ggtggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac
1981 catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg
2041 ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac
2101 caagtggtgg ctatcgccag caacattggc ggcaagcaag cgctcgaaac ggtgcagcgg
2161 ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg accaagtggt ggctatcgcc
2221 agccacgatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc
2281 caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagccacga tggcggcaag
2341 caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc
2401 ccggaccaag tggtggctat cgccagcaac attggcggca agcaagcgct cgaaacggtg
2461 cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct
2521 atcgccagca acattggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg
2581 ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacggtggc
2641 ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc
2701 ctgactccgg accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa
2761 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg
2821 gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg
2881 ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac
2941 ggtggcggca agcaagcgct cgaaagcatt gtggcccagc tgagccggcc tgatccggcg
3001 ttggccgcgt tgaccaacga ccatctggtg gcgttggcat gtcttggtgg acgtcccgcg
3061 ctcgatgcag tcaaaaaggg tctgcctcat gctcccgcat tgatcaaaag aaccaaccgg
3121 cggattcccg agagaacttc ccatcgagtc gcgggatccc aactagtcaa aagtgaactg
3181 gaggagaaga aatctgaact tcgtcataaa ttgaaatatg tgcctcatga atatattgaa
3241 ttaattgaaa ttgccagaaa ttccactcag gatagaattc ttgaaatgaa ggtaatggaa
3301 ttttttatga aagtttatgg atatagaggt aaacatttgg gtggatcaag gaaaccggac
3361 ggagcaattt atactgtcgg atctcctatt gattacggtg tgatcgtgga tactaaagct
3421 tatagcggag gttataatct gccaattggc caagcagatg aaatgcaacg atatgtcgaa
3481 gaaaatcaaa cacgaaacaa acatatcaac cctaatgaat ggtggaaagt ctatccatct
3541 tctgtaacgg aatttaagtt tttatttgtg agtggtcact ttaaaggaaa ctacaaagct
3601 cagcttacac gattaaatca tatcactaat tgtaatggag ctgttcttag tgtagaagag
3661 cttttaattg gtggagaaat gattaaagcc ggcacattaa ccttagagga agtcagacgg
3721 aaatttaata acggcgagat aaactttaga tcttgataac tcgagtctag agggcccgtt
3781 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
3841 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
3901 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
3961 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
4021 tctatggctt ctactgggcg gttttatgga cagcaagcga accggaattg ccagctgggg
4081 cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc tcgccgccaa
4141 ggatctgatg gcgcagggga tcaagctctg atcaagagac aggatgagga tcgtttcgca
4201 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
4261 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
4321 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
4381 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
4441 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
4501 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
4561 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
4621 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
4681 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
4741 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
4801 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
4861 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
4921 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
4981 acgagttctt ctgaattatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
5041 atctgtgcgg tatttcacac cgcatacagg tggcactttt cggggaaatg tgcgcggaac
5101 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
5161 ctgataaatg cttcaataat agcacgtgct aaaacttcat ttttaattta aaaggatcta
5221 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
5281 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
5341 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
5401 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
5461 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
5521 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
5581 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
5641 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
5701 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
5761 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
5821 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
5881 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
5941 gggcttttgc tggccttttg ctcacatgtt ctt
//
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