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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
负20度
- 保质期：
2年
- 供应商：
钦诚生物
- 规格：
5ug
基本信息
| 启动子: | CaMV 35S promoter |
|---|---|
| 复制子: | pUC ori，pVS1 oriV |
| 终止子: | 3A terminator |
| 质粒分类: | 植物系列质粒；植物编辑质粒；植物Cas9质粒 |
| 质粒标签: | C-SV40 NLS |
| 原核抗性: | 壮观霉素Spe |
| 筛选标记: | 潮霉素Hygro，红色荧光蛋白mCherry |
| 克隆菌株: | DH5a等大肠杆菌 |
| 培养条件: | 37℃，有氧，LB |
| 表达宿主: | 拟南芥等植物 |
| 诱导方式: | 无需诱导，组成型表达 |
| 5'测序引物: | 35S:GACGCACAATCCCACTATCC |
| 3'测序引物: | 3A-R：CCATTTGCATTTTGATGTCC |
质粒简介
pHDE-35S-Cas9-mCherry质粒可以在植物中表达Cas9酶和mCherry红色荧光蛋白，可视化基因编辑操作。For CRISPR/Cas9 mediated gene editing in Arabidopsis; provides a visual screen for Cas9-free plants
质粒图谱
质粒序列
LOCUS Exported 16086 bp ds-DNA circular SYN 27-JAN-2018
FEATURES Location/Qualifiers
source 1..16086
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 1..25
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 271..1197
/codon_start=1
/label=mCherry
/translation="MMSGIVLFVTCHSFPTHNPDNNVRASLSLHTHSCMHAFLHVIAMQ
ISFLTYKYKPTLHYTLHSNQNKKTYTNSKTVPTMDNMAIIKEFMRFKVHMEGSVNGHEF
EIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLS
FPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEA
SSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSH
NEDYTIVEQYERAEGRHSTGGMDELYK"
terminator 1203..1455
/label=MAS terminator
/note="mannopine synthase terminator"
promoter 2602..2785
/function="NOS promoter"
/label=NOS promoter
/note="nopaline synthase promoter"
CDS 2837..3862
/codon_start=1
/label=Hyg
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDIGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPGSW
R"
terminator 3900..4152
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
promoter 4285..4630
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS 4635..8738
/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
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 8751..8771
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
misc_feature 9521..9545
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 10073..10864
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=Spe
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 11108..11696
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 11882..12022
/label=bom
/note="basis of mobility region from pBR322"
rep_origin 12366..12560
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(12626..13693)
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
MRYRNLIEGEASAGS"
CDS complement(14127..14756)
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactagtgaa gtgaagcttg
61 gtctcaacta tgagacctaa gctggcacaa ctatatttcc aacatcacta gctaccatca
121 aaagattgac ttctcatctt actcgattga aaccaaatta acatagggtt tttatttaaa
181 taaaagttta accttctttt taaaaaattg ttcatagtgt catgtcagaa caagagctac
241 aaatcacaca tagcatgcat aagcggagct atgatgagtg gtattgtttt gttcgtcact
301 tgtcactctt ttccaacaca taatcccgac aacaacgtaa gagcatctct ctctctccac
361 acacactcat gcatgcatgc attcttacac gtgattgcca tgcaaatctc ctttctcacc
421 tataaataca aaccaaccct tcactacact cttcactcaa accaaaacaa gaaaacatac
481 acaaatagca aaacggtacc aacaatggat aacatggcca tcatcaagga gttcatgcgc
541 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
601 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
661 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
721 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
781 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
841 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
901 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
961 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
1021 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
1081 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
1141 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtgagag
1201 ctcttggact cccatgttgg caaaggcaac caaacaaaca atgaatgatc cgctcctgca
1261 tatggggcgg tttgagtatt tcaactgcca tttgggctga attgaagaca tgctcctgtc
1321 agaaattccg tgatcttact caatattcag taatctcggc caatatccta aatgtgcgtg
1381 gctttatctg tctttgtatt gtttcatcaa ttcatgtaac gtttgctttt cttatgaatt
1441 ttcaaataaa ttatcgtatt gagaccgaat tctcgtcgac gagtcagtaa taaacggcgt
1501 caaagtggtt gcagccggca cacacgagtc gtgtttatca actcaaagca caaatacttt
1561 tcctcaacct aaaaataagg caattagcca aaaacaactt tgcgtgtaaa caacgctcaa
1621 tacacgtgtc attttattat tagctattgc ttcaccgcct tagctttctc gtgacctagt
1681 cgtcctcgtc ttttcttctt cttcttctat aaaacaatac ccaaagagct cttcttcttc
1741 acaattcaga tttcaatttc tcaaaatctt aaaaactttc tctcaattct ctctaccgtg
1801 atcaaggtaa atttctgtgt tccttattct ctcaaaatct tcgattttgt tttcgttcga
1861 tcccaatttc gtatatgttc tttggtttag attctgttaa tcttagatcg aagacgattt
1921 tctgggtttt gatcgttaga tatcatctta attctcgatt agggttttca tagatatcat
1981 ccgatttgtt caaataattt gagttttgtc gaataattac tcttcgattt gtgatttcta
2041 tctagatctg gtgttagttt ctagtttgtg cgatcgaatt tgtcgattaa tctgagtttt
2101 tctgattaac agctcgcaat tgacctaact gagtaagcta gcttgttcga gtattatggc
2161 attgggaaaa ctgtttttct tgtaccattt gttgtgcttg taatttactg tgttttttat
2221 tcggttttcg ctatcgaact gtgaaatgga aatggatgga gaagagttaa tgaatgatat
2281 ggtccttttg ttcattctca aattaatatt atttgttttt tctcttattt gttgtgtgtt
2341 gaatttgaaa ttataagaga tatgcaaaca ttttgttttg agtaaaaatg tgtcaaatcg
2401 tggcctctaa tgaccgaagt taatatgagg agtaaaacac tagatcccca aacaagcttg
2461 aaactgaagg cgggaaacga caatctgatc atgagcggag aattaaggga gtcacgttat
2521 gacccccgcc gatgacgcgg gacaagccgt tttacgtttg gaactgacag aaccgcaacg
2581 ttgaaggagc cactcagccg cgggtttctg gagtttaatg agctaagcac atacgtcaga
2641 aaccattatt gcgcgttcaa aagtcgccta aggtcactat cagctagcaa atatttcttg
2701 tcaaaaatgc tccactgacg ttccataaat tcccctcggt atccaattag agtctcatat
2761 tcactctcaa tccaaataat ctgcaccgga tccgctagag gatctcgacc tgcaagatcc
2821 cggggggcaa tgagatatga aaaagcctga actcaccgcg acgtctgtcg agaagtttct
2881 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
2941 tgctttcagc ttcgatatag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
3001 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc
3061 ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc
3121 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
3181 cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc
3241 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
3301 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
3361 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
3421 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
3481 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
3541 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
3601 gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta
3661 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
3721 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc
3781 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
3841 tcgtccggga tcttggaggt gatgtaacat gatcacaagc tgatcccccg aatttccccg
3901 atcgttcaaa catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga
3961 tgattatcat ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca
4021 tgacgttatt tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg
4081 cgatagaaaa caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta
4141 tgttactaga tcgggaattg atcccccctc gacagcttgc atgccagctt gggctgcagg
4201 tcgacgcgtc aacatggtgg agcacgacac tctcgtctac tccaagaata tcaaagatac
4261 agtctcagaa gaccaaaggg ctattgagac ttttcaacaa agggtaatat cgggaaacct
4321 cctcggattc cattgcccag ctatctgtca cttcatcaaa aggacagtag aaaaggaagg
4381 tggcacctac aaatgccatc attgcgataa aggaaaggct atcgttcaag atgcctctgc
4441 cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt
4501 tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga
4561 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt
4621 ggagaggaca cgatatggac aagaagtact ccattgggct cgatatcggc acaaacagcg
4681 tcggctgggc cgtcattacg gacgagtaca aggtgccgag caaaaaattc aaagttctgg
4741 gcaataccga tcgccacagc ataaagaaga acctcattgg cgccctcctg ttcgactccg
4801 gggagacggc cgaagccacg cggctcaaaa gaacagcacg gcgcagatat acccgcagaa
4861 agaatcggat ctgctacctg caggagatct ttagtaatga gatggctaag gtggatgact
4921 ctttcttcca taggctggag gagtcctttt tggtggagga ggataaaaag cacgagcgcc
4981 acccaatctt tggcaatatc gtggacgagg tggcgtacca tgaaaagtac ccaaccatat
5041 atcatctgag gaagaagctt gtagacagta ctgataaggc tgacttgcgg ttgatctatc
5101 tcgcgctggc gcatatgatc aaatttcggg gacacttcct catcgagggg gacctgaacc
5161 cagacaacag cgatgtcgac aaactcttta tccaactggt tcagacttac aatcagcttt
5221 tcgaagagaa cccgatcaac gcatccggag ttgacgccaa agcaatcctg agcgctaggc
5281 tgtccaaatc ccggcggctc gaaaacctca tcgcacagct ccctggggag aagaagaacg
5341 gcctgtttgg taatcttatc gccctgtcac tcgggctgac ccccaacttt aaatctaact
5401 tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga cacctacgat gatgatctcg
5461 acaatctgct ggcccagatc ggcgaccagt acgcagacct ttttttggcg gcaaagaacc
5521 tgtcagacgc cattctgctg agtgatattc tgcgagtgaa cacggagatc accaaagctc
5581 cgctgagcgc tagtatgatc aagcgctatg atgagcacca ccaagacttg actttgctga
5641 aggcccttgt cagacagcaa ctgcctgaga agtacaagga aattttcttc gatcagtcta
5701 aaaatggcta cgccggatac attgacggcg gagcaagcca ggaggaattt tacaaattta
5761 ttaagcccat cttggaaaaa atggacggca ccgaggagct gctggtaaag cttaacagag
5821 aagatctgtt gcgcaaacag cgcactttcg acaatggaag catcccccac cagattcacc
5881 tgggcgaact gcacgctatc ctcaggcggc aagaggattt ctaccccttt ttgaaagata
5941 acagggaaaa gattgagaaa atcctcacat ttcggatacc ctactatgta ggccccctcg
6001 cccggggaaa ttccagattc gcgtggatga ctcgcaaatc agaagagacc atcactccct
6061 ggaacttcga ggaagtcgtg gataaggggg cctctgccca gtccttcatc gaaaggatga
6121 ctaactttga taaaaatctg cctaacgaaa aggtgcttcc taaacactct ctgctgtacg
6181 agtacttcac agtttataac gagctcacca aggtcaaata cgtcacagaa gggatgagaa
6241 agccagcatt cctgtctgga gagcagaaga aagctatcgt ggacctcctc ttcaagacga
6301 accggaaagt taccgtgaaa cagctcaaag aagactattt caaaaagatt gaatgtttcg
6361 actctgttga aatcagcgga gtggaggatc gcttcaacgc atccctggga acgtatcacg
6421 atctcctgaa aatcattaaa gacaaggact tcctggacaa tgaggagaac gaggacattc
6481 ttgaggacat tgtcctcacc cttacgttgt ttgaagatag ggagatgatt gaagaacgct
6541 tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa acagctcaag aggcgccgat
6601 atacaggatg ggggcggctg tcaagaaaac tgatcaatgg gatccgagac aagcagagtg
6661 gaaagacaat cctggatttt cttaagtccg atggatttgc caaccggaac ttcatgcagt
6721 tgatccatga tgactctctc acctttaagg aggacatcca gaaagcacaa gtttctggcc
6781 agggggacag tcttcacgag cacatcgcta atcttgcagg tagcccagct atcaaaaagg
6841 gaatactgca gaccgttaag gtcgtggatg aactcgtcaa agtaatggga aggcataagc
6901 ccgagaatat cgttatcgag atggcccgag agaaccaaac tacccagaag ggacagaaga
6961 acagtaggga aaggatgaag aggattgaag agggtataaa agaactgggg tcccaaatcc
7021 ttaaggaaca cccagttgaa aacacccagc ttcagaatga gaagctctac ctgtactacc
7081 tgcagaacgg cagggacatg tacgtggatc aggaactgga catcaatcgg ctctccgact
7141 acgacgtgga tcatatcgtg ccccagtctt ttctcaaaga tgattctatt gataataaag
7201 tgttgacaag atccgataaa aatagaggga agagtgataa cgtcccctca gaagaagttg
7261 tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc caaactgatc acacaacgga
7321 agttcgataa tctgactaag gctgaacgag gtggcctgtc tgagttggat aaagccggct
7381 tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa gcacgtggcc caaattctcg
7441 attcacgcat gaacaccaag tacgatgaaa atgacaaact gattcgagag gtgaaagtta
7501 ttactctgaa gtctaagctg gtctcagatt tcagaaagga ctttcagttt tataaggtga
7561 gagagatcaa caattaccac catgcgcatg atgcctacct gaatgcagtg gtaggcactg
7621 cacttatcaa aaaatatccc aagcttgaat ctgaatttgt ttacggagac tataaagtgt
7681 acgatgttag gaaaatgatc gcaaagtctg agcaggaaat aggcaaggcc accgctaagt
7741 acttctttta cagcaatatt atgaattttt tcaagaccga gattacactg gccaatggag
7801 agattcggaa gcgaccactt atcgaaacaa acggagaaac aggagaaatc gtgtgggaca
7861 agggtaggga tttcgcgaca gtccggaagg tcctgtccat gccgcaggtg aacatcgtta
7921 aaaagaccga agtacagacc ggaggcttct ccaaggaaag tatcctcccg aaaaggaaca
7981 gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa gaaatacggc ggattcgatt
8041 ctcctacagt cgcttacagt gtactggttg tggccaaagt ggagaaaggg aagtctaaaa
8101 aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat ggagcgatca agcttcgaaa
8161 aaaaccccat cgactttctc gaggcgaaag gatataaaga ggtcaaaaaa gacctcatca
8221 ttaagcttcc caagtactct ctctttgagc ttgaaaacgg ccggaaacga atgctcgcta
8281 gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc ctctaaatac gttaatttct
8341 tgtatctggc cagccactat gaaaagctca aagggtctcc cgaagataat gagcagaagc
8401 agctgttcgt ggaacaacac aaacactacc ttgatgagat catcgagcaa ataagcgaat
8461 tctccaaaag agtgatcctc gccgacgcta acctcgataa ggtgctttct gcttacaata
8521 agcacaggga taagcccatc agggagcagg cagaaaacat tatccacttg tttactctga
8581 ccaacttggg cgcgcctgca gccttcaagt acttcgacac caccatagac agaaagcggt
8641 acacctctac aaaggaggtc ctggacgcca cactgattca tcagtcaatt acggggctct
8701 atgaaacaag aatcgacctc tctcagctcg gtggagacag cagggctgac cccaagaaga
8761 agaggaaggt gtgatcgatc caggcctccc agctttcgtc cgtatcatcg gtttcgacaa
8821 cgttcgtcaa gttcaatgca tcagtttcat tgcccacaca ccagaatcct actaagtttg
8881 agtattatgg cattggaaaa gctgttttct tctatcattt gttctgcttg taatttactg
8941 tgttctttca gtttttgttt tcggacatca aaatgcaaat ggatggataa gagttaataa
9001 atgatatggt ccttttgttc attctcaaat tattattatc tgttgttttt actttaatgg
9061 gttgaattta agtaagaaag gaactaacag tgtgatatta aggtgcaatg ttagacatat
9121 aaaacagtct ttcacctctc tttggttatg tcttgaattg gtttgtttct tcacttatct
9181 gtgtaatcaa gtttactatg agtctatgat caagtaatta tgcaatcaag ttaagtacag
9241 tataggcttt ttgtgtcgag ggggtaccga gtcgaggaat tcactggccg tcgttttaca
9301 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
9361 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
9421 cagcctgaat ggcgggtacc gagctcgaat tcaattcggc gttaattcag tacattaaaa
9481 acgtccgcaa tgtgttatta agttgtctaa gcgtcaattt gtttacacca caatatatcc
9541 tgccaccagc cagccaacag ctccccgacc ggcagctcgg cacaaaatca ccactcgata
9601 caggcagccc atcagtccgg gacggcgtca gcgggagagc cgttgtaagg cggcagactt
9661 tgctcatgtt accgatgcta ttcggaagaa cggcaactaa gctgccgggt ttgaaacacg
9721 gatgatctcg cggagggtag catgttgatt gtaacgatga cagagcgttg ctgcctgtga
9781 tcaattcggg cacgaaccca gtggacataa gcctcgttcg gttcgtaagc tgtaatgcaa
9841 gtagcgtaac tgccgtcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
9901 ggcgcagtgg cggttttcat ggcttcttgt tatgacatgt ttttttgggg tacagtctat
9961 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
10021 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca tcatggggga
10081 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
10141 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
10201 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
10261 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
10321 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
10381 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
10441 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
10501 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
10561 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
10621 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
10681 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
10741 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
10801 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
10861 ataatgtcta gctagaaatt cgttcaagcc gacgccgctt cgccggcgtt aactcaagcg
10921 attagatgca ctaagcacat aattgctcac agccaaacta tcaggtcaag tctgctttta
10981 ttatttttaa gcgtgcataa taagccctac acaaattggg agatatatca tgcatgacca
11041 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
11101 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
11161 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
11221 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
11281 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
11341 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
11401 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
11461 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
11521 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
11581 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
11641 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
11701 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
11761 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
11821 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
11881 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
11941 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
12001 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
12061 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
12121 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagggtgcct tgatgtgggc
12181 gccggcggtc gagtggcgac ggcgcggctt gtccgcgccc tggtagattg cctggccgta
12241 ggccagccat ttttgagcgg ccagcggccg cgataggccg acgcgaagcg gcggggcgta
12301 gggagcgcag cgaccgaagg gtaggcgctt tttgcagctc ttcggctgtg cgctggccag
12361 acagttatgc acaggccagg cgggttttaa gagttttaat aagttttaaa gagttttagg
12421 cggaaaaatc gccttttttc tcttttatat cagtcactta catgtgtgac cggttcccaa
12481 tgtacggctt tgggttccca atgtacgggt tccggttccc aatgtacggc tttgggttcc
12541 caatgtacgt gctatccaca ggaaagagac cttttcgacc tttttcccct gctagggcaa
12601 tttgccctag catctgctcc gtacattagg aaccggcgga tgcttcgccc tcgatcaggt
12661 tgcggtagcg catgactagg atcgggccag cctgccccgc ctcctccttc aaatcgtact
12721 ccggcaggtc atttgacccg atcagcttgc gcacggtgaa acagaacttc ttgaactctc
12781 cggcgctgcc actgcgttcg tagatcgtct tgaacaacca tctggcttct gccttgcctg
12841 cggcgcggcg tgccaggcgg tagagaaaac ggccgatgcc gggatcgatc aaaaagtaat
12901 cggggtgaac cgtcagcacg tccgggttct tgccttctgt gatctcgcgg tacatccaat
12961 cagctagctc gatctcgatg tactccggcc gcccggtttc gctctttacg atcttgtagc
13021 ggctaatcaa ggcttcaccc tcggataccg tcaccaggcg gccgttcttg gccttcttcg
13081 tacgctgcat ggcaacgtgc gtggtgttta accgaatgca ggtttctacc aggtcgtctt
13141 tctgctttcc gccatcggct cgccggcaga acttgagtac gtccgcaacg tgtggacgga
13201 acacgcggcc gggcttgtct cccttccctt cccggtatcg gttcatggat tcggttagat
13261 gggaaaccgc catcagtacc aggtcgtaat cccacacact ggccatgccg gccggccctg
13321 cggaaacctc tacgtgcccg tctggaagct cgtagcggat cacctcgcca gctcgtcggt
13381 cacgcttcga cagacggaaa acggccacgt ccatgatgct gcgactatcg cgggtgccca
13441 cgtcatagag catcggaacg aaaaaatctg gttgctcgtc gcccttgggc ggcttcctaa
13501 tcgacggcgc accggctgcc ggcggttgcc gggattcttt gcggattcga tcagcggccg
13561 cttgccacga ttcaccgggg cgtgcttctg cctcgatgcg ttgccgctgg gcggcctgcg
13621 cggccttcaa cttctccacc aggtcatcac ccagcgccgc gccgatttgt accgggccgg
13681 atggtttgcg accgtcacgc cgattcctcg ggcttggggg ttccagtgcc attgcagggc
13741 cggcagacaa cccagccgct tacgcctggc caaccgcccg ttcctccaca catggggcat
13801 tccacggcgt cggtgcctgg ttgttcttga ttttccatgc cgcctccttt agccgctaaa
13861 attcatctac tcatttattc atttgctcat ttactctggt agctgcgcga tgtattcaga
13921 tagcagctcg gtaatggtct tgccttggcg taccgcgtac atcttcagct tggtgtgatc
13981 ctccgccggc aactgaaagt tgacccgctt catggctggc gtgtctgcca ggctggccaa
14041 cgttgcagcc ttgctgctgc gtgcgctcgg acggccggca cttagcgtgt ttgtgctttt
14101 gctcattttc tctttacctc attaactcaa atgagttttg atttaatttc agcggccagc
14161 gcctggacct cgcgggcagc gtcgccctcg ggttctgatt caagaacggt tgtgccggcg
14221 gcggcagtgc ctgggtagct cacgcgctgc gtgatacggg actcaagaat gggcagctcg
14281 tacccggcca gcgcctcggc aacctcaccg ccgatgcgcg tgcctttgat cgcccgcgac
14341 acgacaaagg ccgcttgtag ccttccatcc gtgacctcaa tgcgctgctt aaccagctcc
14401 accaggtcgg cggtggccca tatgtcgtaa gggcttggct gcaccggaat cagcacgaag
14461 tcggctgcct tgatcgcgga cacagccaag tccgccgcct ggggcgctcc gtcgatcact
14521 acgaagtcgc gccggccgat ggccttcacg tcgcggtcaa tcgtcgggcg gtcgatgccg
14581 acaacggtta gcggttgatc ttcccgcacg gccgcccaat cgcgggcact gccctgggga
14641 tcggaatcga ctaacagaac atcggccccg gcgagttgca gggcgcgggc tagatgggtt
14701 gcgatggtcg tcttgcctga cccgcctttc tggttaagta cagcgataac cttcatgcgt
14761 tccccttgcg tatttgttta tttactcatc gcatcatata cgcagcgacc gcatgacgca
14821 agctgtttta ctcaaataca catcaccttt ttagacggcg gcgctcggtt tcttcagcgg
14881 ccaagctggc cggccaggcc gccagcttgg catcagacaa accggccagg atttcatgca
14941 gccgcacggt tgagacgtgc gcgggcggct cgaacacgta cccggccgcg atcatctccg
15001 cctcgatctc ttcggtaatg aaaaacggtt cgtcctggcc gtcctggtgc ggtttcatgc
15061 ttgttcctct tggcgttcat tctcggcggc cgccagggcg tcggcctcgg tcaatgcgtc
15121 ctcacggaag gcaccgcgcc gcctggcctc ggtgggcgtc acttcctcgc tgcgctcaag
15181 tgcgcggtac agggtcgagc gatgcacgcc aagcagtgca gccgcctctt tcacggtgcg
15241 gccttcctgg tcgatcagct cgcgggcgtg cgcgatctgt gccggggtga gggtagggcg
15301 ggggccaaac ttcacgcctc gggccttggc ggcctcgcgc ccgctccggg tgcggtcgat
15361 gattagggaa cgctcgaact cggcaatgcc ggcgaacacg gtcaacacca tgcggccggc
15421 cggcgtggtg gtgtcggccc acggctctgc caggctacgc aggcccgcgc cggcctcctg
15481 gatgcgctcg gcaatgtcca gtaggtcgcg ggtgctgcgg gccaggcggt ctagcctggt
15541 cactgtcaca acgtcgccag ggcgtaggtg gtcaagcatc ctggccagct ccgggcggtc
15601 gcgcctggtg ccggtgatct tctcggaaaa cagcttggtg cagccggccg cgtgcagttc
15661 ggcccgttgg ttggtcaagt cctggtcgtc ggtgctgacg cgggcatagc ccagcaggcc
15721 agcggcggcg ctcttgttca tggcgtaatg tctccggttc tagtcgcaag tattctactt
15781 tatgcgacta aaacacgcga caagaaaacg ccaggaaaag ggcagggcgg cagcctgtcg
15841 cgtaacttag gacttgtgcg acatgtcgtt ttcagaagac ggctgcactg aacgtcagaa
15901 gccgactgca ctatagcagc ggaggggttg gatcaaagta ctttaaagta ctttaaagta
15961 ctttaaagta ctttgatccc gaggggaacc ctgtggttgg catgcacata caaatggacg
16021 aacggataaa ccttttcacg cccttttaaa tatccgatta ttctaataaa cgctcttttc
16081 tcttag
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文献和实验相关实验
诺奖得主 Science 发文：基因编辑诞生 10 年，未来将如何改变世界？
前言 簇状规则间隔短回文重复（CRISPR）基因组编辑的出现，以及计算和成像能力的进步，使得编辑细胞和生物体中的 DNA 片段或特定的碱基对成为可能。从遗传病到农业实践和产品，CRISPR 工具箱及其应用深刻地影响了生物学研究。 通过 CRISPR-Cas9，我们不仅可以诊断人类疾病，甚至可以根据个人遗传学预测个体易感性，还可以根据这些信息设计治疗策略。同样，我们既可以识别并快速改变负责植物性状的基因，又可以优化农业研究和植物育种。 2012 年 6 月，Jennifer
-guidedengineerednuclease) 核糖核蛋白 (RGENribonucleoproteins，RGENRNPs) 编辑系统。瞬时表达 CRISPR/Cas9 编辑系统的操作流程和原理是:将 CRISPR/Cas9 质粒 DNA(TECCDNA) 或其转录的 RNA(TECCRNA) 通过基因枪法直接转入植物愈伤组织，因是环状质粒或 RNA，故不易被整合进植物基因组中;在完成切割使命后，质粒 DNA 或 RNA 会被细胞内源核酸酶分解，从而实现全程 DNA-free 的基因组编辑。研究人员对该系统在小麦基因组中的定向修饰特性
，从而诱导 CRISPR 结构通过这些空隙进入细胞中。在电脉冲撤离后，细胞恢复原有状态，这种物理学的方式并不影响细胞内部任何代谢及生理变化。BTX 电转和 CRISPR外源物质如何高效的进入目的细胞是进行表达修饰实验（CRISPR、基因编辑、基因工程）成功的关键。BTX 电穿孔系统由于它的易用性、重复性、高效率和低毒性，已经成为将 CRISPR 结构导入细胞（如哺乳动物细胞、细菌、酵母、植物、寄生虫等）的重要手段。使用 BTX 系统可进行多种类型转染；体外培养--贴壁细胞、悬浮细胞及原代细胞·在体·在卵
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pHDE-35s-Cas9-mCherry植物编辑质粒
¥1000
