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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃
- 保质期：
6个月
- 英文名：
pET-Cas9-NLS-6xHis
- 库存：
大量
- 供应商：
钰博生物
- 规格：
1ug/5ug
pET-Cas9-NLS-6xHis
载体基本信息
| 出品公司: | Ybscience |
|---|---|
| 载体名称: | pET-Cas9-NLS-6xHis |
| 质粒类型: | CRISPR/Cas9载体；细菌基因敲除载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 插入位点: | 限制性内切酶，多克隆位点 |
| 启动子: | lac/T7 |
| 载体大小: | 9265 bp |
| 5' 测序引物及序列: | T7 forward：5’TAATACGACTCACTATAGGG 3’ |
| 3' 测序引物及序列: | T7 Terminal：5’GCTAGTTATTGCTCAGCGG 3’ |
| 载体标签: | NLS 、 His tag (C-ter） |
| 载体抗性: | Ampicillin |
| 克隆菌株: | Mach1 |
| 宿主细胞（系）: | 细菌，如大肠杆菌 |
| 备注: | |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 诱导型 |
| 病毒/非病毒: | 非病毒 |
载体质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS Exported 9265 bp ds-DNA circular SYN 26-JUL-2016
DEFINITION Bacterial vector for regulated expression of S. pyogenes Cas9 with a
C-terminal nuclear localization signal and 6xHis tag.
ACCESSION .
VERSION .
KEYWORDS pET-Cas9-NLS-6xHis
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9265)
AUTHORS Zuris JA, Thompson DB, Shu Y, Guilinger JP, Bessen JL, Hu JH, Maeder
ML, Joung JK, Chen ZY, Liu DR.
TITLE Cationic lipid-mediated delivery of proteins enables efficient
protein-based genome editing in vitro and in vivo.
JOURNAL Nat. Biotechnol. 2015;33:73-80.
PUBMED 25357182
REFERENCE 2 (bases 1 to 9265)
AUTHORS Liu Lab / Addgene #62933
TITLE Direct Submission
JOURNAL Exported Tuesday, July 26, 2016 from SnapGene Viewer 3.1.4
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9265
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
protein_bind 3..27
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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)."
RBS 42..64
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 71..4174
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/note="Cas9"
/note="generates RNA-guided double strand breaks in DNA"
/translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 4175..4195
/codon_start=1
/product="nuclear localization signal of SV40 large T
antigen"
/note="SV40 NLS"
/translation="PKKKRKV"
CDS 4205..4222
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 4307..4354
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 4391..4846
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(4961..5821)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5822..5914)
/gene="bla"
/note="AmpR promoter"
rep_origin 5995..6583
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7013..7204)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(7776..8858)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/note="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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
promoter complement(8859..8936)
/gene="lacI"
/note="lacI promoter"
promoter 9249..2
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
61 gagatatacc atggataaga aatactcaat aggcttagat atcggcacaa atagcgtcgg
121 atgggcggtg atcactgatg aatataaggt tccgtctaaa aagttcaagg ttctgggaaa
181 tacagaccgc cacagtatca aaaaaaatct tataggggct cttttatttg acagtggaga
241 gacagcggaa gcgactcgtc tcaaacggac agctcgtaga aggtatacac gtcggaagaa
301 tcgtatttgt tatctacagg agattttttc aaatgagatg gcgaaagtag atgatagttt
361 ctttcatcga cttgaagagt cttttttggt ggaagaagac aagaagcatg aacgtcatcc
421 tatttttgga aatatagtag atgaagttgc ttatcatgag aaatatccaa ctatctatca
481 tctgcgaaaa aaattggtag attctactga taaagcggat ttgcgcttaa tctatttggc
541 cttagcgcat atgattaagt ttcgtggtca ttttttgatt gagggagatt taaatcctga
601 taatagtgat gtggacaaac tatttatcca gttggtacaa acctacaatc aattatttga
661 agaaaaccct attaacgcaa gtggagtaga tgctaaagcg attctttctg cacgattgag
721 taaatcaaga cgattagaaa atctcattgc tcagctcccc ggtgagaaga aaaatggctt
781 atttgggaat ctcattgctt tgtcattggg tttgacccct aattttaaat caaattttga
841 tttggcagaa gatgctaaat tacagctttc aaaagatact tacgatgatg atttagataa
901 tttattggcg caaattggag atcaatatgc tgatttgttt ttggcagcta agaatttatc
961 agatgctatt ttactttcag atatcctaag agtaaatact gaaataacta aggctcccct
1021 atcagcttca atgattaaac gctacgatga acatcatcaa gacttgactc ttttaaaagc
1081 tttagttcga caacaacttc cagaaaagta taaagaaatc ttttttgatc aatcaaaaaa
1141 cggatatgca ggttatattg atgggggagc tagccaagaa gaattttata aatttatcaa
1201 accaatttta gaaaaaatgg atggtactga ggaattattg gtgaaactaa atcgtgaaga
1261 tttgctgcgc aagcaacgga cctttgacaa cggctctatt ccccatcaaa ttcacttggg
1321 tgagctgcat gctattttga gaagacaaga agacttttat ccatttttaa aagacaatcg
1381 tgagaagatt gaaaaaatct tgacttttcg aattccttat tatgttggtc cattggcgcg
1441 tggcaatagt cgttttgcat ggatgactcg gaagtctgaa gaaacaatta ccccatggaa
1501 ttttgaagaa gttgtcgata aaggtgcttc agctcaatca tttattgaac gcatgacaaa
1561 ctttgataaa aatcttccaa atgaaaaagt actaccaaaa catagtttgc tttatgagta
1621 ttttacggtt tataacgaat tgacaaaggt caaatatgtt actgaaggaa tgcgaaaacc
1681 agcatttctt tcaggtgaac agaagaaagc cattgttgat ttactcttca aaacaaatcg
1741 aaaagtaacc gttaagcaat taaaagaaga ttatttcaaa aaaatagaat gttttgatag
1801 tgttgaaatt tcaggagttg aagatagatt taatgcttca ttaggtacct accatgattt
1861 gctaaaaatt attaaagata aagatttttt ggataatgaa gaaaatgaag atatcttaga
1921 ggatattgtt ttaacattga ccttatttga agatagggag atgattgagg aaagacttaa
1981 aacatatgct cacctctttg atgataaggt gatgaaacag cttaaacgtc gccgttatac
2041 tggttgggga cgtttgtctc gaaaattgat taatggtatt agggataagc aatctggcaa
2101 aacaatatta gattttttga aatcagatgg ttttgccaat cgcaatttta tgcagctgat
2161 ccatgatgat agtttgacat ttaaagaaga cattcaaaaa gcacaagtgt ctggacaagg
2221 cgatagttta catgaacata ttgcaaattt agctggtagc cctgctatta aaaaaggtat
2281 tttacagact gtaaaagttg ttgatgaatt ggtcaaagta atggggcggc ataagccaga
2341 aaatatcgtt attgaaatgg cacgtgaaaa tcagacaact caaaagggcc agaaaaattc
2401 gcgagagcgt atgaaacgaa tcgaagaagg tatcaaagaa ttaggaagtc agattcttaa
2461 agagcatcct gttgaaaata ctcaattgca aaatgaaaag ctctatctct attatctcca
2521 aaatggaaga gacatgtatg tggaccaaga attagatatt aatcgtttaa gtgattatga
2581 tgtcgatcac attgttccac aaagtttcct taaagacgat tcaatagaca ataaggtctt
2641 aacgcgttct gataaaaatc gtggtaaatc ggataacgtt ccaagtgaag aagtagtcaa
2701 aaagatgaaa aactattgga gacaacttct aaacgccaag ttaatcactc aacgtaagtt
2761 tgataattta acgaaagctg aacgtggagg tttgagtgaa cttgataaag ctggttttat
2821 caaacgccaa ttggttgaaa ctcgccaaat cactaagcat gtggcacaaa ttttggatag
2881 tcgcatgaat actaaatacg atgaaaatga taaacttatt cgagaggtta aagtgattac
2941 cttaaaatct aaattagttt ctgacttccg aaaagatttc caattctata aagtacgtga
3001 gattaacaat taccatcatg cccatgatgc gtatctaaat gccgtcgttg gaactgcttt
3061 gattaagaaa tatccaaaac ttgaatcgga gtttgtctat ggtgattata aagtttatga
3121 tgttcgtaaa atgattgcta agtctgagca agaaataggc aaagcaaccg caaaatattt
3181 cttttactct aatatcatga acttcttcaa aacagaaatt acacttgcaa atggagagat
3241 tcgcaaacgc cctctaatcg aaactaatgg ggaaactgga gaaattgtct gggataaagg
3301 gcgagatttt gccacagtgc gcaaagtatt gtccatgccc caagtcaata ttgtcaagaa
3361 aacagaagta cagacaggcg gattctccaa ggagtcaatt ttaccaaaaa gaaattcgga
3421 caagcttatt gctcgtaaaa aagactggga tccaaaaaaa tatggtggtt ttgatagtcc
3481 aacggtagct tattcagtcc tagtggttgc taaggtggaa aaagggaaat cgaagaagtt
3541 aaaatccgtt aaagagttac tagggatcac aattatggaa agaagttcct ttgaaaaaaa
3601 tccgattgac tttttagaag ctaaaggata taaggaagtt aaaaaagact taatcattaa
3661 actacctaaa tatagtcttt ttgagttaga aaacggtcgt aaacggatgc tggctagtgc
3721 cggagaatta caaaaaggaa atgagctggc tctgccaagc aaatatgtga attttttata
3781 tttagctagt cattatgaaa agttgaaggg tagtccagaa gataacgaac aaaaacaatt
3841 gtttgtggag cagcataagc attatttaga tgagattatt gagcaaatca gtgaattttc
3901 taagcgtgtt attttagcag atgccaattt agataaagtt cttagtgcat ataacaaaca
3961 tagagacaaa ccaatacgtg aacaagcaga aaatattatt catttattta cgttgacgaa
4021 tcttggagct cccgctgctt ttaaatattt tgatacaaca attgatcgta aacgatatac
4081 gtctacaaaa gaagttttag atgccactct tatccatcaa tccatcactg gtctttatga
4141 aacacgcatt gatttgagtc agctaggagg tgaccccaag aagaagagga aggtgatgga
4201 taagcatcac caccaccatc actaatgata atttgaacgc cagcacatgg actcgtctac
4261 tagcgcagct taattaacct aggctgctgc caccgctgag caataactag cataacccct
4321 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
4381 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
4441 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
4501 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
4561 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
4621 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
4681 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
4741 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
4801 caaaaattta acgcgaattt taacaaaata ttaacgttta caatttctgg cggcacgatg
4861 gcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
4921 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
4981 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
5041 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
5101 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
5161 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
5221 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
5281 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
5341 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
5401 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
5461 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
5521 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
5581 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
5641 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
5701 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
5761 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
5821 tactcttcct ttttcaatca tgattgaagc atttatcagg gttattgtct catgagcgga
5881 tacatatttg aatgtattta gaaaaataaa caaataggtc atgaccaaaa tcccttaacg
5941 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
6001 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
6061 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
6121 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
6181 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
6241 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
6301 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
6361 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
6421 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
6481 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
6541 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
6601 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
6661 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
6721 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
6781 ttttctcctt acgcatctgt gcggtatttc acaccgcata tatggtgcac tctcagtaca
6841 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
6901 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
6961 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
7021 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
7081 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
7141 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
7201 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
7261 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt
7321 gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc
7381 aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg
7441 cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg
7501 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
7561 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
7621 gccagcctag ccgggtcctc aacgacagga gcacgatcat gctagtcatg ccccgcgccc
7681 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
7741 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
7801 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
7861 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
7921 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
7981 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
8041 tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
8101 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
8161 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
8221 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
8281 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
8341 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
8401 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
8461 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
8521 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
8581 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
8641 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
8701 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
8761 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
8821 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
8881 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
8941 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
9001 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
9061 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
9121 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
9181 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatcg atctcgatcc
9241 cgcgaaatta atacgactca ctata
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文献和实验相关实验
Expression and Purification of Recombinant -Defensins and -Defensin Precursors in Escherichia coli
Recombinant expression of α-defensins can be obtained at efficient levels in Escherichia coli. Amplified α-defensin or pro-α-defensin coding cDNA sequences are cloned directionally between EcoRI and SalI sites of the pET-28a expression vector
到 Cas9(D10A) 的 N 末端，并将核定位信号 (NLS) 肽添加到 Cas9(D10A) 的 C 末端，构建了 APOBEC1-XTEN-Cas9(D10A) 植物基因组单碱基编辑系统。利用该系统对水稻 NRT1.1B 和 SLR1 进行编辑，结果表明该系统能在靶位点产生预期的 C→T(G→A) 碱基替换，NRT1.1B 和 SLR1 在靶位点产生预期突变的效率分别为 2.7% 和 13.3%。夏兰琴和赵云德合作团队也利用 APOBEC1、Cas9 变体 (nCas9-D10A) 和 UGI
改进大家都一样， 1） 优化密码子，让Cas9蛋白可以在哺乳动物系统中很好的表达。 2） 加了核定位序列(NLS)，这样可以把Cas9送到细胞核内进行基因组编辑。 3） 当然，同时需要表达一个guide RNA，把Cas9定位到靶DNA处。 用这个技术，就可以很方便高效廉价地在细胞上做基因编辑：包括敲除、修改、插入。 六：进一步用Nickase增加编辑特异性 因为Cas9介导的靶向主要依赖于guide RNA和目标DNA20左右的碱基的配对，大家开始重视脱靶
技术资料暂无技术资料 索取技术资料
pET-Cas9-NLS-6xHis
¥800 - 1200
