| 质粒名称: | xCas9 3.6 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 108384 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 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: | -- |
| 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: | xCas9 3.6 |
| 物种来源: | Synthetic |
| 插入大小: | 4140 bp |
| 基因突变: | E108G, S217A, A262T, S409I, E480K, E543D, M694I, E1219V |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | NLS（插入件上的N端子） |
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | CMV Forward |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | xCas9 3.6 was a gift from David Liu (Addgene plasmid # 108384 ; http://n2t.net/addgene:108384 ; RRID:Addgene_108384) |
| 参考文献部分: | Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Hu JH, Miller SM, Geurts MH, Tang W, Chen L, Sun N, Zeina CM, Gao X, Rees HA, Lin Z, Liu DR. Nature. 2018 Feb 28. pii: nature26155. doi: 10.1038/nature26155. 10.1038/nature26155 PubMed 29512652 |
买家导航
xCas9 3.6载体 xCas93.6质粒基本信息
xCas9 3.6质粒图谱 xCas93.6载体图谱和xCas9 3.6载体序列 xCas93.6质粒序列多克隆位点信息
xCas9 3.6质粒 xCas93.6载体简介
xCas9 3.6质粒序列 xCas93.6载体序列
LOCUS xCas9 3.6 9553 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Mammalian expression vector for xCas9 3.6.
ACCESSION .
VERSION .
KEYWORDS xCas9 3.6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9553)
AUTHORS Hu JH, Miller SM, Geurts MH, Tang W, Chen L, Sun N, Zeina CM, Gao X,
Rees HA, Lin Z, Liu DR
TITLE Evolved Cas9 variants with broad PAM compatibility and high DNA
specificity.
JOURNAL Nature. 2018 Feb 28. pii: nature26155. doi: 10.1038/nature26155.
PUBMED 29512652
REFERENCE 2 (bases 1 to 9553)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9553
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 50..429
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 430..633
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 584..604
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 630..654
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 742..751
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 748..4851
/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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVGEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLAKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFI
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 4864..4884
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(4939..4958)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4983..5031
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5233..5661
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5320..5339)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5530..5551
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5670..5690)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 5675..6004
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5855..5990
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5917..5936
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 6071..6865
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(6125..6144)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6735..6754
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 7041..7162
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7078..7097)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7132..7151
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(7211..7227)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7211..7227)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7224..7246)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7235..7251
/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(7259..7289)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7304..7325
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7442..7459)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7613..8201)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7693..7712)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8372..9232)
/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 8995..9014
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9233..9337)
/gene="bla"
/label=AmpR promoter
primer_bind complement(9412..9431)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
721 actctagagg atcgaaccct tgccaccatg gacaagaagt actccattgg gctcgatatc
781 ggcacaaaca gcgtcggctg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa
841 ttcaaagttc tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc
901 ctgttcgact ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga
961 tatacccgca gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct
1021 aaggtggatg actctttctt ccataggctg gaggagtcct ttttggtggg ggaggataaa
1081 aagcacgagc gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag
1141 tacccaacca tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg
1201 cggttgatct atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag
1261 ggggacctga acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact
1321 tacaatcagc ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc
1381 ctgagcgcta ggctggccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg
1441 gagaagaaga acggcctgtt tggtaatctt atcgccctgt ccctcgggct gacccccaac
1501 tttaaatcta acttcgacct ggccgaagat accaagcttc aactgagcaa agacacctac
1561 gatgatgatc tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg
1621 gcggcaaaga acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag
1681 atcaccaaag ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac
1741 ttgactttgc tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc
1801 ttcgatcagt ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa
1861 ttttacaaat ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta
1921 aagcttaaca gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aatcatcccc
1981 caccagattc acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc
2041 tttttgaaag ataacaggga aaagattgag aaaatcctca catttcggat accctactat
2101 gtaggccccc tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag
2161 accatcactc cctggaactt cgagaaagtc gtggataagg gggcctctgc ccagtccttc
2221 atcgaaagga tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac
2281 tctctgctgt acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca
2341 gaagggatga gaaagccagc attcctgtct ggagatcaga agaaagctat tgtggacctc
2401 ctcttcaaga cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag
2461 attgaatgtt tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg
2521 ggaacgtatc acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag
2581 aacgaggaca ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg
2641 attgaagaac gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaagcagctc
2701 aaaaggcgcc gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga
2761 gacaagcaga gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg
2821 aacttcattc agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca
2881 caagtttctg gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca
2941 gctatcaaaa agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg
3001 ggaaggcata agcccgagaa tatcgttatc gagatggccc gagagaacca aaccacccag
3061 aagggacaga agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg
3121 gggtcccaaa tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc
3181 tacctgtact acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat
3241 cggctctccg actacgacgt ggaccatatc gtgccccagt cttttctcaa agatgattct
3301 attgataata aagtgttgac aagatccgat aaaaacagag ggaagagtga taacgtcccc
3361 tcagaagaag ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg
3421 atcacacaac ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg
3481 gataaagccg gtttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg
3541 gcccaaattc tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga
3601 gaggtgaaag ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag
3661 ttttataagg tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca
3721 gtggtaggca ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga
3781 gactataaag tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag
3841 gccaccgcta agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca
3901 ctggccaatg gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa
3961 atcgtgtggg acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag
4021 gtgaacatcg ttaaaaagac cgaagtacag accggaggtt tctccaagga aagtatcctc
4081 ccgaaaagga acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac
4141 ggcggattcg attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa
4201 gggaagtcta aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga
4261 tcaagcttcg aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa
4321 aaagacctca tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa
4381 cgaatgctcg ctagtgcggg cgtgctgcag aaaggtaacg agctggcact gccctctaaa
4441 tacgttaatt tcttgtatct ggccagccac tatgaaaagc tcaaggggtc tcccgaagat
4501 aatgagcaga agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag
4561 caaataagcg aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt
4621 tctgcttaca ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac
4681 ttgtttactc tgaccaactt gggcgcgcct gcagccttca agtacttcga cactaccata
4741 gacagaaagc ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca
4801 attacggggc tttatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct
4861 gaccccaaga agaagaggaa ggtgtgaaag ggttcgatcc ctaccggtta gtaatgagtt
4921 taaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
4981 gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg
5041 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
5101 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
5161 tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta
5221 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
5281 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
5341 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
5401 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
5461 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
5521 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
5581 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
5641 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
5701 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
5761 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
5821 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
5881 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
5941 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
6001 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
6061 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
6121 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
6181 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
6241 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
6301 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
6361 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
6421 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
6481 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
6541 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
6601 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
6661 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
6721 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
6781 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
6841 tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag
6901 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
6961 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
7021 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
7081 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
7141 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
7201 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
7261 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
7321 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
7381 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
7441 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
7501 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
7561 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
7621 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
7681 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
7741 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
7801 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
7861 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
7921 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
7981 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
8041 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
8101 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
8161 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
8221 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
8281 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
8341 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
8401 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
8461 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
8521 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
8581 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
8641 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
8701 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
8761 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
8821 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
8881 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
8941 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
9001 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
9061 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
9121 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
9181 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
9241 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
9301 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
9361 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
9421 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
9481 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
9541 gaatctgctt agg
//
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xCas9 3.6质粒 xCas93.6载体应用举例
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