| 质粒名称: | xCas9(3.7)-BE3 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 108380 |
| 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(3.7)-BE3,rAPOBEC1-XTEN-nxCas9(3.7)-UGI-NLS |
| 物种来源: | Synthetic |
| 插入大小: | 5133 bp |
| 基因突变: | A262T, R324L, 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 | M13 |
|
文献引用方法 |
|
| 材料和方法部分: | xCas9(3.7)-BE3 was a gift from David Liu (Addgene plasmid # 108380 ; http://n2t.net/addgene:108380 ; RRID:Addgene_108380) |
| 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(3.7)-BE3载体质粒基本信息
xCas9(3.7)-BE3质粒图谱载体图谱和xCas9(3.7)-BE3载体序列质粒序列多克隆位点信息
-BE3质粒图谱.png)
xCas9(3.7)-BE3质粒载体简介
xCas9(3.7)-BE3质粒序列载体序列
LOCUS xCas9(3.7)-BE3 8532 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Mammalian expression vector for xCas9(3.7)-BE3.
ACCESSION .
VERSION .
KEYWORDS xCas9(3.7)-BE3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8532)
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 8532)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8532
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 74..453
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 454..657
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 608..628
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 699..718
/label=T7
/note="T7 promoter, forward primer"
promoter 699..717
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 731..1417
/codon_start=1
/product="cytidine deaminase (C to U editing enzyme) from
rat"
/label=APOBEC-1
/note="can use ssDNA as a substrate (Komor et al., 2016)"
/translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
LWATGLK"
CDS 1466..5566
/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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5579..5827
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 5840..5860
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5869..5886
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(5909..5926)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5915..6139
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(6210..6226)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6210..6226)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6223..6245)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6234..6250
/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(6258..6288)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6303..6324
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6441..6458)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6612..7200)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6692..6711)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7371..8231)
/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 7994..8013
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8232..8336)
/gene="bla"
/label=AmpR promoter
primer_bind complement(8415..8434)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
61 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
661 ttagtgaacc gtcagatccg ctagagatcc gcggccgcta atacgactca ctatagggag
721 agccgccacc atgagctcag agactggccc agtggctgtg gaccccacat tgagacggcg
781 gatcgagccc catgagtttg aggtattctt cgatccgaga gagctccgca aggagacctg
841 cctgctttac gaaattaatt gggggggccg gcactccatt tggcgacata catcacagaa
901 cactaacaag cacgtcgaag tcaacttcat cgagaagttc acgacagaaa gatatttctg
961 tccgaacaca aggtgcagca ttacctggtt tctcagctgg agcccatgcg gcgaatgtag
1021 tagggccatc actgaattcc tgtcaaggta tccccacgtc actctgttta tttacatcgc
1081 aaggctgtac caccacgctg acccccgcaa tcgacaaggc ctgcgggatt tgatctcttc
1141 aggtgtgact atccaaatta tgactgagca ggagtcagga tactgctgga gaaactttgt
1201 gaattatagc ccgagtaatg aagcccactg gcctaggtat ccccatctgt gggtacgact
1261 gtacgttctt gaactgtact gcatcatact gggcctgcct ccttgtctca acattctgag
1321 aaggaagcag ccacagctga cattctttac catcgctctt cagtcttgtc attaccagcg
1381 actgccccca cacattctct gggccaccgg gttgaaaagc ggcagcgaga ctcccgggac
1441 ctcagagtcc gccacacccg aaagtgacaa gaagtactcc attgggctcg ctatcggcac
1501 aaacagcgtc ggctgggccg tcattacgga cgagtacaag gtgccgagca aaaaattcaa
1561 agttctgggc aataccgatc gccacagcat aaagaagaac ctcattggcg ccctcctgtt
1621 cgactccggg gagacggccg aagccacgcg gctcaaaaga acagcacggc gcagatatac
1681 ccgcagaaag aatcggatct gctacctgca ggagatcttt agtaatgaga tggctaaggt
1741 ggatgactct ttcttccata ggctggagga gtcctttttg gtggaggagg ataaaaagca
1801 cgagcgccac ccaatctttg gcaatatcgt ggacgaggtg gcgtaccatg aaaagtaccc
1861 aaccatatat catctgagga agaagcttgt agacagtact gataaggctg acttgcggtt
1921 gatctatctc gcgctggcgc atatgatcaa atttcgggga cacttcctca tcgaggggga
1981 cctgaaccca gacaacagcg atgtcgacaa actctttatc caactggttc agacttacaa
2041 tcagcttttc gaagagaacc cgatcaacgc atccggagtt gacgccaaag caatcctgag
2101 cgctaggctg tccaaatccc ggcggctcga aaacctcatc gcacagctcc ctggggagaa
2161 gaagaacggc ctgtttggta atcttatcgc cctgtccctc gggctgaccc ccaactttaa
2221 atctaacttc gacctggccg aagataccaa gcttcaactg agcaaagaca cctacgatga
2281 tgatctcgac aatctgctgg cccagatcgg cgaccagtac gcagaccttt ttttggcggc
2341 aaagaacctg tcagacgcca ttctgctgag tgatattctg cgagtgaaca cggagatcac
2401 caaagctccg ctgagcgcta gtatgatcaa gctctatgat gagcaccacc aagacttgac
2461 tttgctgaag gcccttgtca gacagcaact gcctgagaag tacaaggaaa ttttcttcga
2521 tcagtctaaa aatggctacg ccggatacat tgacggcgga gcaagccagg aggaatttta
2581 caaatttatt aagcccatct tggaaaaaat ggacggcacc gaggagctgc tggtaaagct
2641 taacagagaa gatctgttgc gcaaacagcg cactttcgac aatggaatca tcccccacca
2701 gattcacctg ggcgaactgc acgctatcct caggcggcaa gaggatttct accccttttt
2761 gaaagataac agggaaaaga ttgagaaaat cctcacattt cggataccct actatgtagg
2821 ccccctcgcc cggggaaatt ccagattcgc gtggatgact cgcaaatcag aagagaccat
2881 cactccctgg aacttcgaga aagtcgtgga taagggggcc tctgcccagt ccttcatcga
2941 aaggatgact aactttgata aaaatctgcc taacgaaaag gtgcttccta aacactctct
3001 gctgtacgag tacttcacag tttataacga gctcaccaag gtcaaatacg tcacagaagg
3061 gatgagaaag ccagcattcc tgtctggaga tcagaagaaa gctattgtgg acctcctctt
3121 caagacgaac cggaaagtta ccgtgaaaca gctcaaagaa gactatttca aaaagattga
3181 atgtttcgac tctgttgaaa tcagcggagt ggaggatcgc ttcaacgcat ccctgggaac
3241 gtatcacgat ctcctgaaaa tcattaaaga caaggacttc ctggacaatg aggagaacga
3301 ggacattctt gaggacattg tcctcaccct tacgttgttt gaagataggg agatgattga
3361 agaacgcttg aaaacttacg ctcatctctt cgacgacaaa gtcatgaagc agctcaagag
3421 gcgccgatat acaggatggg ggcggctgtc aagaaaactg atcaatggga tccgagacaa
3481 gcagagtgga aagacaatcc tggattttct taagtccgat ggatttgcca accggaactt
3541 cattcagttg atccatgatg actctctcac ctttaaggag gacatccaga aagcacaagt
3601 ttctggccag ggggacagtc ttcacgagca catcgctaat cttgcaggta gcccagctat
3661 caaaaaggga atactgcaga ccgttaaggt cgtggatgaa ctcgtcaaag taatgggaag
3721 gcataagccc gagaatatcg ttatcgagat ggcccgagag aaccaaacca cccagaaggg
3781 acagaagaac agtagggaaa ggatgaagag gattgaagag ggtataaaag aactggggtc
3841 ccaaatcctt aaggaacacc cagttgaaaa cacccagctt cagaatgaga agctctacct
3901 gtactacctg cagaacggca gggacatgta cgtggatcag gaactggaca tcaatcggct
3961 ctccgactac gacgtggatc atatcgtgcc ccagtctttt ctcaaagatg attctattga
4021 taataaagtg ttgacaagat ccgataaaaa cagagggaag agtgataacg tcccctcaga
4081 agaagttgtc aagaaaatga aaaattattg gcggcagctg ctgaacgcca aactgatcac
4141 acaacggaag ttcgataatc tgactaaggc tgaacgaggt ggcctgtctg agttggataa
4201 agccggtttc atcaaaaggc agcttgttga gacacgccag atcaccaagc acgtggccca
4261 aattctcgat tcacgcatga acaccaagta cgatgaaaat gacaaactga ttcgagaggt
4321 gaaagttatt actctgaagt ctaagctggt ctcagatttc agaaaggact ttcagtttta
4381 taaggtgaga gagatcaaca attaccacca tgcgcatgat gcctacctga atgcagtggt
4441 aggcactgca cttatcaaaa aatatcccaa gcttgaatct gaatttgttt acggagacta
4501 taaagtgtac gatgttagga aaatgatcgc aaagtctgag caggaaatag gcaaggccac
4561 cgctaagtac ttcttttaca gcaatattat gaattttttc aagaccgaga ttacactggc
4621 caatggagag attcggaagc gaccacttat cgaaacaaac ggagaaacag gagaaatcgt
4681 gtgggacaag ggtagggatt tcgcgacagt ccggaaggtc ctgtccatgc cgcaggtgaa
4741 catcgttaaa aagaccgaag tacagaccgg aggcttctcc aaggaaagta tcctcccgaa
4801 aaggaacagc gacaagctga tcgcacgcaa aaaagattgg gaccccaaga aatacggcgg
4861 attcgattct cctacagtcg cttacagtgt actggttgtg gctaaagtgg agaaagggaa
4921 gtctaaaaaa ctcaaaagcg tcaaggaact gctgggcatc acaatcatgg agcgatcaag
4981 cttcgaaaaa aaccccatcg actttctcga ggcgaaagga tataaagagg tcaaaaaaga
5041 cctcatcatt aagcttccca agtactctct ctttgagctt gaaaacggcc ggaaacgaat
5101 gctcgctagt gcgggcgtgc tgcagaaagg taacgagctg gcactgccct ctaaatacgt
5161 taatttcttg tatctggcca gccactatga aaagctcaaa gggtctcccg aagataatga
5221 gcagaagcag ctgttcgtgg aacaacacaa acactacctt gatgagatca tcgagcaaat
5281 aagcgaattc tccaaaagag tgatcctcgc cgacgctaac ctcgataagg tgctttctgc
5341 ttacaataag cacagggata agcccatcag ggagcaggca gaaaacatta tccacttgtt
5401 tactctgacc aacttgggcg cgcctgcagc cttcaagtac ttcgacacta ccatagacag
5461 aaagcggtac acctctacaa aggaggtcct ggacgccaca ctgattcatc agtcaattac
5521 ggggctctat gaaacaagaa tcgacctctc tcagctcggt ggagactctg gtggttctac
5581 taatctgtca gatattattg aaaaggagac cggtaagcaa ctggttatcc aggaatccat
5641 cctcatgctc ccagaggagg tggaagaagt cattgggaac aagccggaaa gcgatatact
5701 cgtgcacacc gcctacgacg agagcaccga cgagaatgtc atgcttctga ctagcgacgc
5761 ccctgaatac aagccttggg ctctggtcat acaggatagc aacggtgaga acaagattaa
5821 gatgctctct ggtggttctc ccaagaagaa gaggaaagtc taaccggtca tcatcaccat
5881 caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
5941 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
6001 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
6061 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
6121 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc gataccgtcg
6181 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
6241 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctagggtgcc
6301 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
6361 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
6421 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
6481 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
6541 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
6601 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
6661 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
6721 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
6781 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
6841 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
6901 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
6961 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
7021 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
7081 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
7141 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
7201 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
7261 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
7321 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
7381 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
7441 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
7501 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
7561 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
7621 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
7681 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
7741 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
7801 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
7861 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
7921 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
7981 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
8041 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
8101 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
8161 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
8221 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
8281 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
8341 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatctcc cgatccccta
8401 gggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
8461 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
8521 ggcttgaccg ac
//
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xCas9(3.7)-BE3质粒载体应用举例
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