| 质粒名称: | pCS2-3xFLAG-NLS-SpCas9-NLS |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 51307 |
| 存储实验室: | Yonglong Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pCS2+ |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4095 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | CRISPR;热带爪蟾表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | 3xFLAG-NLS-SpCas9-NLS |
| 物种来源: | -- |
| 插入大小: | 4272 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: |
3x标志(插入N端子)
NLS(插入N端子)
NLS(插入的C端子)
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | XhoI (未破坏) |
| 5' 测序引物 | SP6 |
| 3' 测序引物 | T3 |
|
文献引用方法 |
|
| 材料和方法部分: | pCS2-3xFLAG-NLS-SpCas9-NLS was a gift from Yonglong Chen (Addgene plasmid # 51307 ; http://n2t.net/addgene:51307 ; RRID:Addgene_51307) |
| 参考文献部分: | Efficient RNA/Cas9-mediated genome editing in Xenopus tropicalis. Guo X, Zhang T, Hu Z, Zhang Y, Shi Z, Wang Q, Cui Y, Wang F, Zhao H, Chen Y. Development. 2014 Jan 8. 10.1242/dev.099853 PubMed 24401372 |
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pCS2-3xFLAG-NLS-SpCas9-NLS载体质粒基本信息
pCS2-3xFLAG-NLS-SpCas9-NLS质粒图谱载体图谱和pCS2-3xFLAG-NLS-SpCas9-NLS载体序列质粒序列多克隆位点信息

pCS2-3xFLAG-NLS-SpCas9-NLS质粒载体简介
pCS2-3xFLAG-NLS-SpCas9-NLS质粒序列载体序列
LOCUS pCS2-3xFLAG-NLS-SpCas9-NLS 8357 bp ds-DNA circular SYN 04-JUN-2019
DEFINITION Cas9 expression plasmid.
ACCESSION .
VERSION .
KEYWORDS pCS2-3xFLAG-NLS-SpCas9-NLS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8357)
AUTHORS Guo X, Zhang T, Hu Z, Zhang Y, Shi Z, Wang Q, Cui Y, Wang F, Zhao H,
Chen Y
TITLE Efficient RNA/Cas9-mediated genome editing in Xenopus tropicalis.
JOURNAL Development. 2014 Jan 8.
PUBMED 24401372
REFERENCE 2 (bases 1 to 8357)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jun 4, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8357
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 112..1089
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
promoter 1124..1142
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1124..1141
/label=SP6
/note="SP6 promoter, forward primer"
CDS 1192..1257
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 1264..1284
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1309..5409
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5410..5457
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
promoter complement(5476..5493)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 5498..5632
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5522..5541)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 5576..5595
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(5745..5765)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(5745..5763)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5784..5800)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5784..5800)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5797..5819)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5808..5824
/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(5832..5862)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5877..5898
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6015..6032)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6186..6774)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6266..6285)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6945..7805)
/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 7568..7587
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7806..7910)
/gene="bla"
/label=AmpR promoter
rep_origin complement(7936..34)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(8073..8094)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 8285..8304
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
61 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagtcgac cgccaattca
121 atatggcgta tatggactca tgccaattca atatggtgga tctggacctg tgccaattca
181 atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc agccaattca
241 atatggcgga cttggcacca tgccaattca atatggcgga cctggcactg tgccaactgg
301 ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc
361 cgccatattg aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc
421 aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata
481 ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat
541 agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag
601 ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg
661 gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg
721 cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc
781 tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc
841 cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag
901 tacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat
961 ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt
1021 ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt
1081 ttagggaacc gccattctgc ctggggacgt cggagcaagc ttgatttagg tgacactata
1141 gaatacaagc tacttgttct ttttgcagga tcccatcgat tcgaattcat ggactataag
1201 gaccacgacg gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg
1261 gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac
1321 agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
1381 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
1441 aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag
1501 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
1561 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
1621 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
1681 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
1741 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
1801 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
1861 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
1921 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg
1981 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gcaacctgat tgccctgagc
2041 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
2101 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
2161 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
2221 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac
2281 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
2341 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc
2401 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
2461 accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc
2521 gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
2581 caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
2641 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
2701 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
2761 gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
2821 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc
2881 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
2941 aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
3001 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
3061 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
3121 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
3181 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
3241 aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag
3301 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
3361 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
3421 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
3481 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
3541 gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga
3601 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
3661 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
3721 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
3781 caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc
3841 tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc
3901 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag
3961 ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga
4021 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
4081 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
4141 aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
4201 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
4261 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa
4321 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
4381 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
4441 ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca
4501 aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa
4561 gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
4621 agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac
4681 tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg
4741 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg
4801 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
4861 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
4921 ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa
4981 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
5041 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac
5101 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
5161 aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag
5221 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
5281 tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc
5341 accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
5401 ggaggcgaca agcgtcctgc tgctactaag aaagctggtc aagctaagaa aaagaaatga
5461 ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat
5521 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
5581 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
5641 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc
5701 ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta
5761 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
5821 acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga
5881 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
5941 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
6001 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
6061 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
6121 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
6181 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
6241 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
6301 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
6361 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
6421 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
6481 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
6541 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
6601 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
6661 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
6721 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
6781 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
6841 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
6901 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
6961 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
7021 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
7081 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
7141 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
7201 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
7261 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
7321 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
7381 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
7441 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
7501 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
7561 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
7621 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
7681 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
7741 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
7801 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
7861 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
7921 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
7981 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
8041 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
8101 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
8161 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
8221 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
8281 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
8341 tgcgcgtaac caccaca
//
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pCS2-3xFLAG-NLS-SpCas9-NLS质粒载体应用举例
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