| 质粒名称: | pPlatTET-gRNA2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 82559 |
| 存储实验室: | Izuho Hatada |
|
载体骨架基本信息 |
|
| 载体名称: | pCAG |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 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: | dCas9-5xPlat2AflD-P2A-scFvGCN4sfGFPTET1CD |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 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 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pPlatTET-gRNA2 was a gift from Izuho Hatada (Addgene plasmid # 82559 ; http://n2t.net/addgene:82559 ; RRID:Addgene_82559) |
| 参考文献部分: | Targeted DNA demethylation in vivo using dCas9-peptide repeat and scFv-TET1 catalytic domain fusions. Morita S, Noguchi H, Horii T, Nakabayashi K, Kimura M, Okamura K, Sakai A, Nakashima H, Hata K, Nakashima K, Hatada I. Nat Biotechnol. 2016 Aug 29. doi: 10.1038/nbt.3658. 10.1038/nbt.3658 PubMed 27571369 |
买家导航
pPlatTET-gRNA2载体 pPlatTETgRNA2质粒基本信息
pPlatTET-gRNA2质粒图谱 pPlatTETgRNA2载体图谱和pPlatTET-gRNA2载体序列 pPlatTETgRNA2质粒序列多克隆位点信息
pPlatTET-gRNA2质粒 pPlatTETgRNA2载体简介
pPlatTET-gRNA2质粒序列 pPlatTETgRNA2载体序列
LOCUS pPlatTET-gRNA2 14290 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION All in one vector which contains Cas9 peptide array (linker length:
22aa), antibody-sfGFP-TET1CD, and gRNA expression system..
ACCESSION .
VERSION .
KEYWORDS pPlatTET-gRNA2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14290)
AUTHORS Morita S, Noguchi H, Horii T, Nakabayashi K, Kimura M, Okamura K,
Sakai A, Nakashima H, Hata K, Nakashima K, Hatada I
TITLE Targeted DNA demethylation in vivo using dCas9-peptide repeat and
scFv-TET1 catalytic domain fusions.
JOURNAL Nat Biotechnol. 2016 Aug 29. doi: 10.1038/nbt.3658.
PUBMED 27571369
REFERENCE 2 (bases 1 to 14290)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..14290
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..304
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
intron 582..1598
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1522..1544
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1606..1625
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 1674..1694
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
regulatory 1743..1752
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1749..5852
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=dCas9
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the H840A mutation in the HNH
catalytic domain"
/translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 5856..5882
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 5901..5921
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5928..5948
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6012..6068
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 6135..6191
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 6258..6314
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 6381..6437
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 6504..6560
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 6648..6704
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 7452..7478
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 7569..8279
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
VTAAGITHGMDELYK"
primer_bind complement(7596..7624)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 8219..8239
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
polyA_signal 10606..10687
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(10643..10662)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 10697..10716
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 10804..10824
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 10975..10994
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
protein_bind complement(11101..11125)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
rep_origin complement(11146..11601)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(11283..11304)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 11495..11514
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 11628..11732
/gene="bla"
/label=AmpR promoter
promoter 11734..12091
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(11757..11777)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 11942..12077
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 12004..12023
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 12126..12920
/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(12180..12199)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 12790..12809
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(13108..13127)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 13152..13199
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 13528..14116
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 14017..14036
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
301 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc
361 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg
421 gggggggcgc gcccggcggg ggggggcggg gcgaggggcg gggcggggcg aggcggagag
481 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
541 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
601 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
661 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct
721 tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg
781 agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga
841 gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc
901 tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg
961 gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag
1021 ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg
1081 agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc
1141 cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg
1201 agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga
1261 gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc
1321 aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg
1381 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
1441 gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc
1501 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct
1561 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
1621 ttgtgctgtc tcatcatttt ggcaaagaat tcccccgcga tcgcgccacc atgcccaaga
1681 agaagcgcaa ggtgggacgc gtctgcagga tatcaagctt gcggtaccgc gggcccggga
1741 tcgccaccat ggacaagaag tacagcatcg gcctggccat cggcaccaac tctgtgggct
1801 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
1861 ccgaccggca cagcatcaag aagaacctga tcggcgccct gctgttcgac agcggagaaa
1921 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
1981 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
2041 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
2101 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
2161 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
2221 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
2281 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
2341 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
2401 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
2461 tcggcaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
2521 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
2581 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
2641 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
2701 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
2761 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
2821 gctacgccgg ctacatcgat ggcggagcca gccaggaaga gttctacaag ttcatcaagc
2881 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
2941 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
3001 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
3061 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
3121 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
3181 tcgaggaagt ggtggacaag ggcgccagcg cccagagctt catcgagcgg atgaccaact
3241 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
3301 tcaccgtgta caacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
3361 ccttcctgag cggcgagcag aaaaaagcca tcgtggacct gctgttcaag accaaccgga
3421 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
3481 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
3541 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
3601 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
3661 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
3721 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
3781 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
3841 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
3901 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
3961 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
4021 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
4081 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
4141 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
4201 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
4261 tggacgctat cgtgcctcag agctttctga aggacgactc catcgataac aaagtgctga
4321 ctcggagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
4381 agatgaagaa ctactggcgc cagctgctga atgccaagct gattacccag aggaagttcg
4441 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
4501 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
4561 ggatgaacac taagtacgac gagaacgaca aactgatccg ggaagtgaaa gtgatcaccc
4621 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
4681 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
4741 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
4801 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
4861 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
4921 ggaagcggcc tctgatcgag acaaacggcg aaacaggcga gatcgtgtgg gataagggcc
4981 gggactttgc caccgtgcgg aaagtgctgt ctatgcccca agtgaatatc gtgaaaaaga
5041 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgaca
5101 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca
5161 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
5221 agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc
5281 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
5341 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
5401 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
5461 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
5521 ttgtggaaca gcacaaacac tacctggacg agatcatcga gcagatcagc gagttctcca
5581 agagagtgat cctggccgac gctaatctgg acaaggtgct gagcgcctac aacaagcaca
5641 gagacaagcc tatcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
5701 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
5761 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
5821 cacggatcga cctgtctcag ctgggaggcg acgcctatcc ctatgacgtg cccgattatg
5881 ccagcctggg cagcggctcc cccaagaaaa aacgcaaggt ggaagatcct aagaaaaagc
5941 ggaaagtgga cggcattggt agtgggagca acggcagcag cggatccaac ggtccgactg
6001 acgccgcgga agaggagctt ctgagcaaaa actatcacct cgaaaacgag gttgcgcgac
6061 tgaagaaagg aagcgggtcc ggtggaagtg gctccggatc tggaggttct ggcagcggag
6121 gtagcggcag tggcgaagag ctccttagta agaactatca tctggaaaat gaggtagcgc
6181 gcttaaagaa agggtcggga agtggcggca gcggaagtgg gagtggaggg agcggttctg
6241 gcggttccgg cagtggagag gagttgctgt ctaagaacta ccacttagaa aacgaagtcg
6301 cacggctaaa aaaaggttcc ggctccggcg gctccggttc tggaagcggg ggctcgggat
6361 caggtggatc tggatcagga gaggaattgc tttccaaaaa ctaccacctt gagaatgagg
6421 tggccaggtt aaagaagggg agcggctcgg ggggtagtgg atcggggtcg ggcgggtcag
6481 gaagcggtgg tagcggatct ggggaggagc tgctctcgaa gaattaccat ttggagaacg
6541 aagtggcgag actaaagaag ggaagcggta gtggtggttc agggtctggt tcaggtggca
6601 gtgggtctgg gggctcaggg tccgggacgg ccggcctcgg aagcggagct actaacttca
6661 gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctagtacc atgggccccg
6721 acatcgtgat gacccagagc cccagcagcc tgagcgccag cgtgggcgac cgcgtgacca
6781 tcacctgccg cagcagcacc ggcgccgtga ccaccagcaa ctacgccagc tgggtgcagg
6841 agaagcccgg caagctgttc aagggcctga tcggcggcac caacaaccgc gcccccggcg
6901 tgcccagccg cttcagcggc agcctgatcg gcgacaaggc caccctgacc atcagcagcc
6961 tgcagcccga ggacttcgcc acctacttct gcgccctgtg gtacagcaac cactgggtgt
7021 tcggccaggg caccaaggtg gagctgaagc gcggcggcgg cggcagcggc ggcggcggca
7081 gcggcggcgg cggcagcagc ggcggcggca gcgaggtgaa gctgctggag agcggcggcg
7141 gcctggtgca gcccggcggc agcctgaagc tgagctgcgc cgtgagcggc ttcagcctga
7201 ccgactacgg cgtgaactgg gtgcgccagg cccccggccg cggcctggag tggatcggcg
7261 tgatctgggg cgacggcatc accgactaca acagcgccct gaaggaccgc ttcatcatca
7321 gcaaggacaa cggcaagaac accgtgtacc tgcagatgag caaggtgcgc agcgacgaca
7381 ccgccctgta ctactgcgtg accggcctgt tcgactactg gggccagggc accctggtga
7441 ccgtgagcag ctacccatac gatgttccag attacgctgg tggaggcgga ggttctgggg
7501 gaggaggtag tggcggtggt ggttcaggag gcggcggaag cttggatcca ggtggaggtg
7561 gaagcggtag caaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag
7621 atggtgatgt taatgggcac aaattttctg tccgtggaga gggtgaaggt gatgctacaa
7681 acggaaaact cacccttaaa tttatttgca ctactggaaa actacctgtt ccgtggccaa
7741 cacttgtcac tactctgacc tatggtgttc aatgcttttc ccgttatccg gatcacatga
7801 aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa cgcactatat
7861 ctttcaaaga tgacgggacc tacaagacgc gtgctgaagt caagtttgaa ggtgataccc
7921 ttgttaatcg tatcgagtta aagggtattg attttaaaga agatggaaac attcttggac
7981 acaaactcga gtacaacttt aactcacaca atgtatacat cacggcagac aaacaaaaga
8041 atggaatcaa agctaacttc aaaattcgcc acaacgttga agatggttcc gttcaactag
8101 cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc
8161 attacctgtc gacacaatct gtcctttcga aagatcccaa cgaaaagcgt gaccacatgg
8221 tccttcttga gtttgtaact gctgctggga ttacacatgg catggatgag ctctacaaag
8281 gtggaggtcg gaccggtggc ggtggcggag gggctagcag atccgaactg cccacctgca
8341 gctgtcttga tcgagttata caaaaagaca aaggcccata ttatacacac cttggggcag
8401 gaccaagtgt tgctgctgtc agggaaatca tggagaatag gtatggtcaa aaaggaaacg
8461 caataaggat agaaatagta gtgtacaccg gtaaagaagg gaaaagctct catgggtgtc
8521 caattgctaa gtgggtttta agaagaagca gtgatgaaga aaaagttctt tgtttggtcc
8581 ggcagcgtac aggccaccac tgtccaactg ctgtgatggt ggtgctcatc atggtgtggg
8641 atggcatccc tcttccaatg gccgaccggc tatacacaga gctcacagag aatctaaagt
8701 catacaatgg gcaccctacc gacagaagat gcaccctcaa tgaaaatcgt acctgtacat
8761 gtcaaggaat tgatccagag acttgtggag cttcattctc ttttggctgt tcatggagta
8821 tgtactttaa tggctgtaag tttggtagaa gcccaagccc cagaagattt agaattgatc
8881 caagctctcc cttacatgaa aaaaaccttg aagataactt acagagtttg gctacacgat
8941 tagctccaat ttataagcag tatgctccag tagcttacca aaatcaggtg gaatatgaaa
9001 atgttgcccg agaatgtcgg cttggcagca aggaaggtcg tcccttctct ggggtcactg
9061 cttgcctgga cttctgtgct catccccaca gggacattca caacatgaat aatggaagca
9121 ctgtggtttg taccttaact cgagaagata accgctcttt gggtgttatt cctcaagatg
9181 agcagctcca tgtgctacct ctttataagc tttcagacac agatgagttt ggctccaagg
9241 aaggaatgga agccaagatc aaatctgggg ccatcgaggt cctggcaccc cgccgcaaaa
9301 aaagaacgtg tttcactcag cctgttcccc gttctggaaa gaagagggct gcgatgatga
9361 cagaggttct tgcacataag ataagggcag tggaaaagaa acctattccc cgaatcaagc
9421 ggaagaataa ctcaacaaca acaaacaaca gtaagccttc gtcactgcca accttaggga
9481 gtaacactga gaccgtgcaa cctgaagtaa aaagtgaaac cgaaccccat tttatcttaa
9541 aaagttcaga caacactaaa acttattcgc tgatgccatc cgctcctcac ccagtgaaag
9601 aggcatctcc aggcttctcc tggtccccga agactgcttc agccacacca gctccactga
9661 agaatgacgc aacagcctca tgcgggtttt cagaaagaag cagcactccc cactgtacga
9721 tgccttcggg aagactcagt ggtgccaatg cagctgctgc tgatggccct ggcatttcac
9781 agcttggcga agtggctcct ctccccaccc tgtctgctcc tgtgatggag cccctcatta
9841 attctgagcc ttccactggt gtgactgagc cgctaacgcc tcatcagcca aaccaccagc
9901 cctccttcct cacctctcct caagaccttg cctcttctcc aatggaagaa gatgagcagc
9961 attctgaagc agatgagcct ccatcagacg aacccctatc tgatgacccc ctgtcacctg
10021 ctgaggagaa attgccccac attgatgagt attggtcaga cagtgagcac atctttttgg
10081 atgcaaatat tggtggggtg gccatcgcac ctgctcacgg ctcggttttg attgagtgtg
10141 cccggcgaga gctgcacgct accactcctg ttgagcaccc caaccgtaat catccaaccc
10201 gcctctccct tgtcttttac cagcacaaaa acctaaataa gccccaacat ggttttgaac
10261 taaacaagat taagtttgag gctaaagaag ctaagaataa gaaaatgaag gcctcagagc
10321 aaaaagacca ggcagctaat gaaggtccag aacagtcctc tgaagtaaat gaattgaacc
10381 aaattccttc tcataaagca ttaacattaa cccatgacaa tgttgtcacc gtgtcccctt
10441 atgctctcac acacgttgcg gggccctata accattgggt ctgagcggcc gcgactctag
10501 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
10561 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
10621 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
10681 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttggcg cgcctgtaca
10741 aaaaagcagg ctttaaagga accaattcag tcgactggat ccggtaccaa ggtcgggcag
10801 gaagagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
10861 gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
10921 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
10981 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttaagttaa
11041 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttctag
11101 acccagcttt cttgtacaaa gttggcatta ggcgcgccaa ggcgtaaatt gtaagcgtta
11161 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
11221 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
11281 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
11341 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
11401 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
11461 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
11521 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
11581 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
11641 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
11701 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
11761 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
11821 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
11881 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
11941 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
12001 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
12061 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
12121 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
12181 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
12241 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
12301 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
12361 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
12421 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
12481 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
12541 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
12601 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
12661 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
12721 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
12781 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
12841 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
12901 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
12961 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
13021 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
13081 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
13141 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
13201 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
13261 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
13321 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
13381 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
13441 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
13501 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
13561 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
13621 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
13681 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
13741 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
13801 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
13861 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
13921 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
13981 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
14041 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
14101 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
14161 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
14221 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
14281 tggagttccg
//
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pPlatTET-gRNA2质粒 pPlatTETgRNA2载体应用举例
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