| 质粒名称: | nls-NgAgo-GK |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 78253 |
| 存储实验室: | Chunyu Han |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4000 bp |
| 完整质粒大小: | 6679 bp |
| 载体修饰: | pEGFP-N1 without the eGFP |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 未知 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | NgAgo |
| 物种来源: | Natronobacterium gregoryi |
| 插入大小: | 2712 bp |
| 基因又名: | Argonaute |
| GenBank编号 | AFZ73749.1 |
| 启动子 | CMV |
| 标签/融合蛋白: | a nuclear localization signal (NLS) (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | HindIII(未破坏) |
| 3' 克隆位点: | BamHI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | nls-NgAgo-GK was a gift from Chunyu Han (爱迪基因 plasmid # 78253) |
| 参考文献部分: | DNA-guided genome editing using the Natronobacterium gregoryi Argonaute. Gao F, Shen XZ, Jiang F, Wu Y, Han C. Nat Biotechnol. 2016 May 2. doi: 10.1038/nbt.3547. 10.1038/nbt.3547 PubMed 27136078 |
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 nls-NgAgo-GK 6679 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Mammalian expression of NgAgo (Argonaute from N. gregoryi)..
ACCESSION .
VERSION .
KEYWORDS nls-NgAgo-GK
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6679)
AUTHORS Gao F, Shen XZ, Jiang F, Wu Y, Han C
TITLE DNA-guided genome editing using the Natronobacterium gregoryi
Argonaute.
JOURNAL Nat Biotechnol. 2016 May 2. doi: 10.1038/nbt.3547.
PUBMED 27136078
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|nls-NgAgo-GK|
FEATURES Location/Qualifiers
source 1..6679
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 16..319
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 320..523
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 474..494
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 583..592
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 595..615
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 619..3276
/codon_start=1
/product="Argonaute protein from Natronobacterium gregoryi
(Gao et al., 2016)"
/label=NgAgo
/note="reportedly a DNA-guided endonuclease that does not
require a PAM site, although this interpretation has been
challenged (Cyranoski, 2016)"
/translation="TVIDLDSTTTADELTSGHTYDISVTLTGVYDNTDEQHPRMSLAFE
QDNGERRYITLWKNTTPKDVFTYDYATGSTYIFTNIDYEVKDGYENLTATYQTTVENAT
AQEVGTTDEDETFAGGEPLDHHLDDALNETPDDAETESDSGHVMTSFASRDQLPEWTLH
TYTLTATDGAKTDTEYARRTLAYTVRQELYTDHDAAPVATDGLMLLTPEPLGETPLDLD
CGVRVEADETRTLDYTTAKDRLLARELVEEGLKRSLWDDYLVRGIDEVLSKEPVLTCDE
FDLHERYDLSVEVGHSGRAYLHINFRHRFVPKLTLADIDDDNIYPGLRVKTTYRPRRGH
IVWGLRDECATDSLNTLGNQSVVAYHRNNQTPINTDLLDAIEAADRRVVETRRQGHGDD
AVSFPQELLAVEPNTHQIKQFASDGFHQQARSKTRLSASRCSEKAQAFAERLDPVRLNG
STVEFSSEFFTGNNEQQLRLLYENGESVLTFRDGARGAHPDETFSKGIVNPPESFEVAV
VLPEQQADTCKAQWDTMADLLNQAGAPPTRSETVQYDAFSSPESISLNVAGAIDPSEVD
AAFVVLPPDQEGFADLASPTETYDELKKALANMGIYSQMAYFDRFRDAKIFYTRNVALG
LLAAAGGVAFTTEHAMPGDADMFIGIDVSRSYPEDGASGQINIAATATAVYKDGTILGH
SSTRPQLGEKLQSTDVRDIMKNAILGYQQVTGESPTHIVIHRDGFMNEDLDPATEFLNE
QGVEYDIVEIRKQPQTRLLAVSDVQYDTPVKSIAAINQNEPRATVATFGAPEYLATRDG
GGLPRPIQIERVAGETDIETLTRQVYLLSQSHIQVHNSTARLPITTAYADQASTHATKG
YLVQTGAFESNVGFL"
polyA_signal 3422..3543
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3459..3478)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3513..3532
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3550..4005)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3687..3708)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3899..3918
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4032..4136
/gene="bla"
/label=AmpR promoter
promoter 4138..4495
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(4161..4181)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4346..4481
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4408..4427
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4530..5324
/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(4584..4603)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5194..5213
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5512..5531)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5556..5603
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5932..6520
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6421..6440
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
541 gatccgctag cgctaccgga ctcagatctc gagctcaagc ttgccaccat ggtgccaaaa
601 aagaagagaa aggtagccac agtgattgac ctcgattcga ccaccaccgc agacgaactg
661 acatcgggac acacgtacga catctcagtc acgctcaccg gtgtctacga taacaccgac
721 gagcagcatc ctcgcatgtc tctcgcattc gagcaggaca acggcgagcg gcgttacatt
781 accctgtgga agaacacgac acccaaggat gtctttacat acgactacgc cacgggctcg
841 acgtacatct tcactaacat cgactacgaa gtgaaggacg gctacgagaa tctgactgca
901 acataccaga cgaccgtcga gaacgctacc gctcaggaag tcgggacgac tgacgaggac
961 gaaacgttcg cgggcggcga gccgctcgac catcacttgg acgacgcgct caatgagacg
1021 ccagacgacg cggagacaga gagcgactca ggccatgtga tgacctcgtt cgcctcccgc
1081 gaccaactcc ctgagtggac gctgcatacg tatacgctaa cagccacaga cggcgcaaag
1141 acggacacgg agtacgcgcg acgaaccctc gcatacacgg tacggcagga actctatacc
1201 gaccatgatg cggctccggt tgcaactgac gggctaatgc ttctcacgcc agagccgctc
1261 ggcgagaccc cgcttgacct cgattgcggt gtccgggtcg aggcggacga gactcggaca
1321 ctcgattaca ccacggccaa agaccggtta ctcgcccgcg aactcgtcga agaggggctc
1381 aaacgctccc tctgggatga ctacctcgtt cgcggcatcg atgaagtcct ctcaaaggag
1441 cctgtgctga cttgcgatga gttcgaccta catgagcggt atgacctctc tgtcgaagtc
1501 ggtcacagtg ggcgggcgta ccttcacatc aacttccgcc accggttcgt accgaagctg
1561 acgctcgcag acatcgatga tgacaacatc tatcctgggc tccgggtgaa gacgacgtat
1621 cgcccccggc gaggacatat cgtctggggt ctgcgggacg agtgcgccac cgactcgctc
1681 aacacgctgg gaaaccagtc cgtcgttgca taccaccgca acaatcagac acctattaac
1741 actgacctcc tcgacgctat cgaggccgct gaccggcgag tcgtcgaaac ccgacgtcaa
1801 gggcacggcg atgatgctgt ctcattcccc caagaactgc ttgcggtcga accgaatacg
1861 caccaaatta agcagttcgc ctccgacgga ttccaccaac aggcccgctc aaagacgcgt
1921 ctctcggcct cccgctgcag cgagaaagcg caagcgttcg ccgagcggct tgacccggtg
1981 cgtctcaatg ggtccacggt agagttctcc tcggagtttt tcaccgggaa caacgagcag
2041 caactgcgcc tcctctacga gaacggtgag tcggttctga cgttccgcga cggggcgcgt
2101 ggtgcgcacc ccgacgagac attctcgaaa ggtatcgtca atccaccaga gtcgttcgag
2161 gtggccgtag tactgcccga gcagcaggca gatacctgca aagcgcagtg ggacacgatg
2221 gctgacctcc tcaaccaagc tggcgcgcca ccgacacgga gcgagaccgt ccaatatgat
2281 gcgttctcct cgccagagag catcagcctc aatgtggctg gagccatcga ccctagcgag
2341 gtagacgcgg cattcgtcgt actgccgccg gaccaagaag gattcgcaga cctcgccagt
2401 ccgacagaga cgtacgacga gctgaagaag gcgcttgcca acatgggcat ttacagccag
2461 atggcgtact tcgaccggtt ccgcgacgcg aaaatattct atactcgtaa cgtggcactc
2521 gggctgctgg cagccgctgg cggcgtcgca ttcacaaccg aacatgcgat gcctggggac
2581 gcagatatgt tcattgggat tgatgtctct cggagctacc ccgaggacgg tgccagcggc
2641 cagataaaca ttgccgcgac ggcgaccgcc gtctacaagg atggaactat cctcggccac
2701 tcgtccaccc gaccgcagct cggggagaaa ctacagtcga cggatgttcg tgacattatg
2761 aagaatgcca tcctcggcta ccagcaggtg accggtgagt cgccgaccca tatcgtcatc
2821 caccgtgatg gcttcatgaa cgaagacctc gaccccgcca cggaattcct caacgaacaa
2881 ggcgtcgagt acgacatcgt cgaaatccgc aagcagcccc agacacgcct gctggcagtc
2941 tccgatgtgc agtacgatac gcctgtgaag agcatcgccg ctatcaacca gaacgagcca
3001 cgggcaacgg tcgccacctt cggcgcaccc gaatacttag cgacacgcga tggaggcggc
3061 cttccccgcc caatccaaat tgaacgagtc gccggcgaaa ccgacatcga gacgctcact
3121 cgccaagtct atctgctctc ccagtcgcat atccaggtcc ataactcgac tgcgcgccta
3181 cccatcacca ccgcatacgc cgaccaggca agtactcacg cgaccaaggg ttacctcgtc
3241 cagaccggag cgttcgagtc taatgtcgga ttcctccggg atccatatgt aagtaagtaa
3301 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
3361 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
3421 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
3481 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
3541 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
3601 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
3661 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
3721 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
3781 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
3841 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
3901 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
3961 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
4021 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
4081 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
4141 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
4201 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
4261 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
4321 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
4381 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
4441 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
4501 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
4561 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
4621 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
4681 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
4741 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
4801 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
4861 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
4921 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
4981 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
5041 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
5101 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
5161 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
5221 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
5281 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
5341 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
5401 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
5461 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
5521 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
5581 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
5641 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
5701 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
5761 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
5821 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
5881 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
5941 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
6001 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
6061 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
6121 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
6181 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
6241 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
6301 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
6361 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
6421 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
6481 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
6541 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
6601 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
6661 ggtcattagt tcatagccc
//
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