质粒名称: | pLCRISPR-CMV |
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出品公司: | 爱迪基因 |
目录编号: | 102609 |
存储实验室: | Nicolas Manel |
载体骨架基本信息 |
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载体名称: | pLCRISPR |
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: | 哺乳动物表达,慢病毒 |
筛选标记: | -- |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 30 ℃ |
插入基因信息 |
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插入基因名称: | -- |
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 |
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材料和方法部分: | pLCRISPR-CMV was a gift from Nicolas Manel (Addgene plasmid # 102609 ; http://n2t.net/addgene:102609 ; RRID:Addgene_102609) |
参考文献部分: | Transmission of innate immune signaling by packaging of cGAMP in viral particles. Gentili M, Kowal J, Tkach M, Satoh T, Lahaye X, Conrad C, Boyron M, Lombard B, Durand S, Kroemer G, Loew D, Dalod M, Thery C, Manel N. Science. 2015 Sep 11;349(6253):1232-6. doi: 10.1126/science.aab3628. Epub 2015 Jul 30. 10.1126/science.aab3628 PubMed 26229115 |
买家导航
pLCRISPRCMV载体基本信息
pLCRISPRCMV载体图谱质粒图谱和多克隆位点信息
pLCRISPRCMV载体简介
pLCRISPRCMV载体序列
LOCUS pLCRISPR-CMV 13834 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Control lentivector expressing Cas9 and gRNA scaffold. ACCESSION . VERSION . KEYWORDS pLCRISPR-CMV SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13834) AUTHORS Gentili M, Kowal J, Tkach M, Satoh T, Lahaye X, Conrad C, Boyron M, Lombard B, Durand S, Kroemer G, Loew D, Dalod M, Thery C, Manel N TITLE Transmission of innate immune signaling by packaging of cGAMP in viral particles. JOURNAL Science. 2015 Sep 11;349(6253):1232-6. doi: 10.1126/science.aab3628. Epub 2015 Jul 30. PUBMED 26229115 REFERENCE 2 (bases 1 to 13834) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13834 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(17..36) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 208..587 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 589..787 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" LTR 805..985 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1032..1157 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1650..1883 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." promoter 2367..2607 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 2367..2387 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 2538..2557 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 2634..2709 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 2743..2860 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 2955..3258 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3259..3462 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" regulatory 3516..3525 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3525..3590 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 3597..3617 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3642..7742 /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 7752..7799 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 7809..7865 /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 7872..8471 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(7872..7891) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 8368..8388 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 8496..9084 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(8549..8569) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(8967..8978) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(9087..9103) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9088..9104) /label=pBluescriptKS /note="For pBluescript vector" LTR 9609..9789 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(9815..9832) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 9821..10045 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 10091..10519 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(10178..10197) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 10388..10409 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(10528..10548) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 10533..10862 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 10713..10848 /label=SV40 ori /note="SV40 origin of replication" primer_bind 10775..10794 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 10910..10957 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 10976..11350 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 11480..11601 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(11517..11536) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 11571..11590 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(11650..11666) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(11650..11666) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(11663..11685) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 11674..11690 /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(11698..11728) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11743..11764 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(11881..11898) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(12052..12640) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(12132..12151) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(12811..13671) /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 13434..13453 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(13672..13776) /gene="bla" /label=AmpR promoter ORIGIN 1 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc 61 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc 121 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc 181 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat 241 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg 301 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt 361 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac 421 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg 481 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact 541 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt 601 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc 661 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc 721 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata 781 taagcagcgc gttttgcctg tactgggtct ctctggttag accagatctg agcctgggag 841 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt 901 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt 961 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga 1021 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg 1081 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag 1141 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc 1201 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg 1261 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa 1321 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata 1381 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag 1441 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg 1501 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa 1561 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg 1621 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca 1681 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg 1741 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg 1801 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga 1861 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc 1921 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac 1981 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta 2041 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa 2101 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc 2161 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg 2221 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg 2281 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc 2341 attcgattag tgaacggatc ggcactgagg gcctatttcc catgattcct tcatatttgc 2401 atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga 2461 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa 2521 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct 2581 tggctttata tatcttgtgg aaaggacgaa acaccgggtc ttcgagaaga cctgttttag 2641 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg 2701 agtcggtgct ttttttagac aaatggcagt attcatccac aattttaaaa gaaaaggggg 2761 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac 2821 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag 2881 cagagatcca gtttggttaa ttaatagtaa tcaattacgg ggtcattagt tcatagccca 2941 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac 3001 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact 3061 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa 3121 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg 3181 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta 3241 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg 3301 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg 3361 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg 3421 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag 3481 atcccgcggg ctagcggatc cccaccggtc gtacggccac catggactat aaggaccacg 3541 acggagacta caaggatcat gatattgatt acaaagacga tgacgataag atggccccaa 3601 agaagaagcg gaaggtcggt atccacggag tcccagcagc cgacaagaag tacagcatcg 3661 gcctggacat cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc 3721 ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga 3781 tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg 3841 ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca 3901 acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg 3961 aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct 4021 accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca 4081 aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact 4141 tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc 4201 tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg 4261 ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc 4321 agctgcccgg cgagaagaag aatggcctgt tcggcaacct gattgccctg agcctgggcc 4381 tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca 4441 aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg 4501 acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag 4561 tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc 4621 accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca 4681 aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca 4741 gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg 4801 aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg 4861 gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag 4921 atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca 4981 tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa 5041 agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg 5101 cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc 5161 tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga 5221 aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca 5281 tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact 5341 acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca 5401 acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg 5461 acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg 5521 acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga 5581 tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca 5641 acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct 5701 tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca 5761 tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg 5821 ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg 5881 tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc 5941 agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca 6001 tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga 6061 acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac 6121 tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag agctttctga 6181 aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg 6241 acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga 6301 acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc 6361 tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca 6421 caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca 6481 agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga 6541 aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct 6601 acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt 6661 tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg 6721 aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga 6781 ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg 6841 aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga 6901 gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag 6961 agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc 7021 ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca 7081 aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca 7141 tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca 7201 aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa 7261 acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc 7321 tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct 7381 cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg 7441 agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg 7501 acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga 7561 atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg 7621 acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga 7681 tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg 7741 acgcggccgc caagcgtcct gctgctacta agaaagctgg tcaagctaag aaaaagaaag 7801 gaagcggagc tactaacttc agcctgctga agcaggctgg agacgtggag gagaaccctg 7861 gacctggatc catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc 7921 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg 7981 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg 8041 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga 8101 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg 8161 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc 8221 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca 8281 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc 8341 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca 8401 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc 8461 ccggtgccta agaattcgat atcaagctta tcgataatca acctctggat tacaaaattt 8521 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg 8581 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt 8641 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg 8701 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc 8761 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg 8821 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt 8881 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc 8941 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg 9001 gcctgctgcc ggctctgcgg cctcttccgc atcttcgcct tcgccctcag acgagtcgga 9061 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgagaccta gaaaaacatg 9121 gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac 9181 aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt 9241 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa 9301 ttcactccca acgaagtcaa gatatccttg atctgtggat ctaccacaca caaggctact 9361 tccctgattg gcagaactac acaccagggc cagggatcag atatccactg acctttggat 9421 ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat gaaggagaga 9481 acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat 9541 tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg 9601 actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag 9661 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc 9721 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa 9781 tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag 9841 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact 9901 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt 9961 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagataa tagcaggcat 10021 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg 10081 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 10141 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 10201 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 10261 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 10321 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 10381 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 10441 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 10501 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc 10561 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 10621 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 10681 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 10741 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 10801 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 10861 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta 10921 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc 10981 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt 11041 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt 11101 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac 11161 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt 11221 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg 11281 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga 11341 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg 11401 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 11461 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa 11521 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 11581 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc 11641 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 11701 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 11761 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 11821 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 11881 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 11941 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 12001 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 12061 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 12121 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 12181 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 12241 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 12301 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 12361 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 12421 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 12481 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 12541 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 12601 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 12661 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 12721 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 12781 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 12841 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 12901 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 12961 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 13021 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 13081 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 13141 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 13201 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 13261 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 13321 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 13381 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 13441 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 13501 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 13561 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 13621 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 13681 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 13741 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 13801 tgacgtcgac ggatcgggag atctcccgat cccc //
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pLCRISPRCMV载体质粒应用举例
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