| 质粒名称: | pLCRISPR-CMV |
|---|---|
| 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 |
|
| 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 |
|
| 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 |
|
| 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
pLCRISPR-CMV载体 pLCRISPRCMV质粒基本信息
pLCRISPR-CMV质粒图谱 pLCRISPRCMV载体图谱和pLCRISPR-CMV载体序列 pLCRISPRCMV质粒序列多克隆位点信息
pLCRISPR-CMV质粒 pLCRISPRCMV载体简介
pLCRISPR-CMV质粒序列 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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pLCRISPR-CMV质粒 pLCRISPRCMV载体应用举例
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