| 质粒名称: | pH-PABE-7-esgRNA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 115620 |
| 存储实验室: | Caixia Gao |
|
载体骨架基本信息 |
|
| 载体名称: | pCAMBIA1300 |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 植物表达，CRISPR |
| 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: | wtTadA-TadA7.10-nCas9-3*NLS |
| 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: | 限制性内切酶 |
| 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: | pH-PABE-7-esgRNA was a gift from Caixia Gao (Addgene plasmid # 115620 ; http://n2t.net/addgene:115620 ; RRID:Addgene_115620) |
| 参考文献部分: | Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion. Li C, Zong Y, Wang Y, Jin S, Zhang D, Song Q, Zhang R, Gao C. Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z. 10.1186/s13059-018-1443-z PubMed 29807545 |
买家导航
pH-PABE-7-esgRNA载体质粒基本信息
pH-PABE-7-esgRNA质粒图谱载体图谱和pH-PABE-7-esgRNA载体序列质粒序列多克隆位点信息
pH-PABE-7-esgRNA质粒载体简介
pH-PABE-7-esgRNA质粒序列载体序列
LOCUS pH-PABE-7-esgRNA 18869 bp ds-DNA circular SYN 11-JAN-2019
DEFINITION Targeted A to G in rice.
ACCESSION .
VERSION .
KEYWORDS pH-PABE-7-esgRNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 18869)
AUTHORS Li C, Zong Y, Wang Y, Jin S, Zhang D, Song Q, Zhang R, Gao C
TITLE Expanded base editing in rice and wheat using a Cas9-adenosine
deaminase fusion.
JOURNAL Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z.
PUBMED 29807545
REFERENCE 2 (bases 1 to 18869)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jan 11, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..18869
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 261..285
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
primer_bind 473..495
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 487..504
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 488..504
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 571..950
/label=OsU3 promoter
/note="Oryza sativa (rice) snRNA U3 promoter"
terminator 1058..1077
/label=SUP4 terminator
/note="transcription terminator for the S. cerevisiae SUP4
tRNA gene"
promoter 1090..3080
/label=Ubi promoter
/note="maize polyubiquitin gene promoter"
CDS 4288..8388
/codon_start=1
/product="nickase mutant of the Cas9 endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system"
/label=Cas9(D10A)
/note="generates RNA-guided single strand nicks in DNA due
to the D10A mutation in the RuvC catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
RBS 8301..8309
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 8401..8421
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 8434..8454
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 8467..8487
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(9146..9162)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9146..9162)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(9159..9181)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 9170..9186
/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(9194..9224)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9239..9260
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9451..10127
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
primer_bind 10064..10084
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
CDS 10194..11219
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
polyA_signal 11260..11434
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
misc_feature 11512..11536
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 11961..12755
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
rep_origin 14180..14768
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 14669..14688
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 14922..14939
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 14954..15094
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(15020..15039)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 15180..15202
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
rep_origin 15438..15632
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(15698..16771)
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
CDS complement(17200..17829)
/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 gcgacaagaa aacgccagga aaagggcagg gcggcagcct gtcgcgtaac ttaggacttg
61 tgcgacatgt cgttttcaga agacggctgc actgaacgtc agaagccgac tgcactatag
121 cagcggaggg gttggatcaa agtactttga tcccgagggg aaccctgtgg ttggcatgca
181 catacaaatg gacgaacgga taaacctttt cacgcccttt taaatatccg attattctaa
241 taaacgctct tttctcttag gtttacccgc caatatatcc tgtcaaacac tgatagttta
301 aactgaaggc gggaaacgac aatctgatcc aagctcaagc tgctctagca ttcgccattc
361 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
421 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
481 cgacgttgta aaacgacggc cagtgccaag cttagtaatt catccaggtc accaagttct
541 aggattttca gaactgcaac ttattttatc aaggaatctt taaacatacg aacagatcac
601 ttaaagttct tctgaagcaa cttaaagtta tcaggcatgc atggatcttg gaggaatcag
661 atgtgcagtc agggaccata gcacaagaca ggcgtcttct actggtgcta ccagcaaatg
721 ctggaagccg ggaacactgg gtacgttgga aaccacgtga tgtgaagaag taagataaac
781 tgtaggagaa aagcatttcg tagtgggcca tgaagccttt caggacatgt attgcagtat
841 gggccggccc attacgcaat tggacgacaa caaagactag tattagtacc acctcggcta
901 tccacataga tcaaagctga tttaaaagag ttgtgcagat gatccgtggc gtgagaccct
961 gcagggtctc tgtttaagag ctatgctgga aacagcatag caagtttaaa taaggctagt
1021 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttgttttt tatgtctaag
1081 cttgcatgcc tgcagtgcag cgtgacccgg tcgtgcccct ctctagagat aatgagcatt
1141 gcatgtctaa gttataaaaa attaccacat attttttttg tcacacttgt ttgaagtgca
1201 gtttatctat ctttatacat atatttaaac tttactctac gaataatata atctatagta
1261 ctacaataat atcagtgttt tagagaatca tataaatgaa cagttagaca tggtctaaag
1321 gacaattgag tattttgaca acaggactct acagttttat ctttttagtg tgcatgtgtt
1381 ctcctttttt tttgcaaata gcttcaccta tataatactt catccatttt attagtacat
1441 ccatttaggg tttagggtta atggttttta tagactaatt tttttagtac atctatttta
1501 ttctatttta gcctctaaat taagaaaact aaaactctat tttagttttt ttatttaata
1561 atttagatat aaaatagaat aaaataaagt gactaaaaat taaacaaata ccctttaaga
1621 aattaaaaaa actaaggaaa catttttctt gtttcgagta gataatgcca gcctgttaaa
1681 cgccgtcgac gagtctaacg gacaccaacc agcgaaccag cagcgtcgcg tcgggccaag
1741 cgaagcagac ggcacggcat ctctgtcgct gcctctggac ccctctcgag agttccgctc
1801 caccgttgga cttgctccgc tgtcggcatc cagaaattgc gtggcggagc ggcagacgtg
1861 agccggcacg gcaggcggcc tcctcctcct ctcacggcac cggcagctac gggggattcc
1921 tttcccaccg ctccttcgct ttcccttcct cgcccgccgt aataaataga caccccctcc
1981 acaccctctt tccccaacct cgtgttgttc ggagcgcaca cacacacaac cagatctccc
2041 ccaaatccac ccgtcggcac ctccgcttca aggtacgccg ctcgtcctcc ccccccccct
2101 ctctaccttc tctagatcgg cgttccggtc catggttagg gcccggtagt tctacttctg
2161 ttcatgtttg tgttagatcc gtgtttgtgt tagatccgtg ctgctagcgt tcgtacacgg
2221 atgcgacctg tacgtcagac acgttctgat tgctaacttg ccagtgtttc tctttgggga
2281 atcctgggat ggctctagcc gttccgcaga cgggatcgat ttcatgattt tttttgtttc
2341 gttgcatagg gtttggtttg cccttttcct ttatttcaat atatgccgtg cacttgtttg
2401 tcgggtcatc ttttcatgct tttttttgtc ttggttgtga tgatgtggtc tggttgggcg
2461 gtcgttctag atcggagtag aattctgttt caaactacct ggtggattta ttaattttgg
2521 atctgtatgt gtgtgccata catattcata gttacgaatt gaagatgatg gatggaaata
2581 tcgatctagg ataggtatac atgttgatgc gggttttact gatgcatata cagagatgct
2641 ttttgttcgc ttggttgtga tgatgtggtg tggttgggcg gtcgttcatt cgttctagat
2701 cggagtagaa tactgtttca aactacctgg tgtatttatt aattttggaa ctgtatgtgt
2761 gtgtcataca tcttcatagt tacgagttta agatggatgg aaatatcgat ctaggatagg
2821 tatacatgtt gatgtgggtt ttactgatgc atatacatga tggcatatgc agcatctatt
2881 catatgctct aaccttgagt acctatctat tataataaac aagtatgttt tataattatt
2941 ttgatcttga tatacttgga tgatggcata tgcagcagct atatgtggat ttttttagcc
3001 ctgccttcat acgctattta tttgcttggt actgtttctt ttgtcgatgc tcaccctgtt
3061 gtttggtgtt acttctgcag ccctaggcct atgtccgagg tggagttctc tcacgagtat
3121 tggatgaggc acgctcttac acttgctaag agagcttggg acgaaagaga agtgccagtt
3181 ggcgccgttc ttgtgcataa taatagggtg atcggcgagg gttggaatag accaattgga
3241 aggcatgatc caacagctca cgcagagatt atggctctca gacaaggcgg cctcgttatg
3301 cagaactaca ggctcattga cgctacactc tacgtgacac tcgaaccttg cgttatgtgc
3361 gccggagcta tgattcattc taggattggc agggtcgtgt ttggagctag ggacgctaaa
3421 acaggagccg ccggatctct tatggacgtg ttgcatcatc caggcatgaa ccatagggtg
3481 gagattacag agggcattct tgcagacgag tgcgctgctc ttctttccga tttcttcagg
3541 atgagaaggc aggagattaa ggcccagaag aaggctcagt cttctacaga tagcggagga
3601 tcttccggag gatctagcgg ctccgagaca ccaggaacat ccgaaagcgc tacaccagaa
3661 tctagcggag gctcttccgg aggatcttct gaagtggagt tctcccacga gtattggatg
3721 aggcacgctc ttacacttgc taaaagggct agggacgaaa gggaagttcc agttggagct
3781 gttctcgtgc tcaataacag ggtgattggc gagggttgga atagagccat tggactccat
3841 gatccaacag ctcacgcaga gattatggct cttagacaag gcggcctcgt tatgcagaat
3901 tacagactca tcgacgccac actctacgtt accttcgaac cttgcgttat gtgcgccgga
3961 gctatgatcc attctaggat tggcagggtc gtgttcggcg ttagaaacgc taagacagga
4021 gctgcaggct ctcttatgga cgttcttcat tacccaggca tgaatcatag agtggagatc
4081 acagaaggca ttcttgcaga cgagtgcgca gctctccttt gctatttctt caggatgccg
4141 aggcaagttt tcaacgctca gaagaaggcc cagtcttcta cagattccgg cggatcttct
4201 ggaggatcta gcggctccga gacaccagga acatccgaat ccgctacacc agagtcttct
4261 ggaggatcta gcggaggatc tcttaaggac aagaagtact cgatcggcct cgccattggg
4321 actaactctg ttggctgggc cgtgatcacc gacgagtaca aggtgccctc aaagaagttc
4381 aaggtcctgg gcaacaccga tcggcattcc atcaagaaga atctcattgg cgctctcctg
4441 ttcgacagcg gcgagacggc tgaggctacg cggctcaagc gcaccgcccg caggcggtac
4501 acgcgcagga agaatcgcat ctgctacctg caggagattt tctccaacga gatggcgaag
4561 gttgacgatt ctttcttcca caggctggag gagtcattcc tcgtggagga ggataagaag
4621 cacgagcggc atccaatctt cggcaacatt gtcgacgagg ttgcctacca cgagaagtac
4681 cctacgatct accatctgcg gaagaagctc gtggactcca cagataaggc ggacctccgc
4741 ctgatctacc tcgctctggc ccacatgatt aagttcaggg gccatttcct gatcgagggg
4801 gatctcaacc cggacaatag cgatgttgac aagctgttca tccagctcgt gcagacgtac
4861 aaccagctct tcgaggagaa ccccattaat gcgtcaggcg tcgacgcgaa ggctatcctg
4921 tccgctaggc tctcgaagtc tcggcgcctc gagaacctga tcgcccagct gccgggcgag
4981 aagaagaacg gcctgttcgg gaatctcatt gcgctcagcc tggggctcac gcccaacttc
5041 aagtcgaatt tcgatctcgc tgaggacgcc aagctgcagc tctccaagga cacatacgac
5101 gatgacctgg ataacctcct ggcccagatc ggcgatcagt acgcggacct gttcctcgct
5161 gccaagaatc tgtcggacgc catcctcctg tctgatattc tcagggtgaa caccgagatt
5221 acgaaggctc cgctctcagc ctccatgatc aagcgctacg acgagcacca tcaggatctg
5281 accctcctga aggcgctggt caggcagcag ctccccgaga agtacaagga gatcttcttc
5341 gatcagtcga agaacggcta cgctgggtac attgacggcg gggcctctca ggaggagttc
5401 tacaagttca tcaagccgat tctggagaag atggacggca cggaggagct gctggtgaag
5461 ctcaatcgcg aggacctcct gaggaagcag cggacattcg ataacggcag catcccacac
5521 cagattcatc tcggggagct gcacgctatc ctgaggaggc aggaggactt ctaccctttc
5581 ctcaaggata accgcgagaa gatcgagaag attctgactt tcaggatccc gtactacgtc
5641 ggcccactcg ctaggggcaa ctcccgcttc gcttggatga cccgcaagtc agaggagacg
5701 atcacgccgt ggaacttcga ggaggtggtc gacaagggcg ctagcgctca gtcgttcatc
5761 gagaggatga cgaatttcga caagaacctg ccaaatgaga aggtgctccc taagcactcg
5821 ctcctgtacg agtacttcac agtctacaac gagctgacta aggtgaagta tgtgaccgag
5881 ggcatgagga agccggcttt cctgtctggg gagcagaaga aggccatcgt ggacctcctg
5941 ttcaagacca accggaaggt cacggttaag cagctcaagg aggactactt caagaagatt
6001 gagtgcttcg attcggtcga gatctctggc gttgaggacc gcttcaacgc ctccctgggg
6061 acctaccacg atctcctgaa gatcattaag gataaggact tcctggacaa cgaggagaat
6121 gaggatatcc tcgaggacat tgtgctgaca ctcactctgt tcgaggaccg ggagatgatc
6181 gaggagcgcc tgaagactta cgcccatctc ttcgatgaca aggtcatgaa gcagctcaag
6241 aggaggaggt acaccggctg ggggaggctg agcaggaagc tcatcaacgg cattcgggac
6301 aagcagtccg ggaagacgat cctcgacttc ctgaagagcg atggcttcgc gaaccgcaat
6361 ttcatgcagc tgattcacga tgacagcctc acattcaagg aggatatcca gaaggctcag
6421 gtgagcggcc agggggactc gctgcacgag catatcgcga acctcgctgg ctcgccagct
6481 atcaagaagg ggattctgca gaccgtgaag gttgtggacg agctggtgaa ggtcatgggc
6541 aggcacaagc ctgagaacat cgtcattgag atggcccggg agaatcagac cacgcagaag
6601 ggccagaaga actcacgcga gaggatgaag aggatcgagg agggcattaa ggagctgggg
6661 tcccagatcc tcaaggagca cccggtggag aacacgcagc tgcagaatga gaagctctac
6721 ctgtactacc tccagaatgg ccgcgatatg tatgtggacc aggagctgga tattaacagg
6781 ctcagcgatt acgacgtcga tcatatcgtt ccacagtcat tcctgaagga tgactccatt
6841 gacaacaagg tcctcaccag gtcggacaag aaccggggca agtctgataa tgttccttca
6901 gaggaggtcg ttaagaagat gaagaactac tggcgccagc tcctgaatgc caagctgatc
6961 acgcagcgga agttcgataa cctcacaaag gctgagaggg gcgggctctc tgagctggac
7021 aaggcgggct tcatcaagag gcagctggtc gagacacggc agatcactaa gcacgttgcg
7081 cagattctcg actcacggat gaacactaag tacgatgaga atgacaagct gatccgcgag
7141 gtgaaggtca tcaccctgaa gtcaaagctc gtctccgact tcaggaagga tttccagttc
7201 tacaaggttc gggagatcaa caattaccac catgcccatg acgcgtacct gaacgcggtg
7261 gtcggcacag ctctgatcaa gaagtaccca aagctcgaga gcgagttcgt gtacggggac
7321 tacaaggttt acgatgtgag gaagatgatc gccaagtcgg agcaggagat tggcaaggct
7381 accgccaagt acttcttcta ctctaacatt atgaatttct tcaagacaga gatcactctg
7441 gccaatggcg agatccggaa gcgccccctc atcgagacga acggcgagac gggggagatc
7501 gtgtgggaca agggcaggga tttcgcgacc gtcaggaagg ttctctccat gccacaagtg
7561 aatatcgtca agaagacaga ggtccagact ggcgggttct ctaaggagtc aattctgcct
7621 aagcggaaca gcgacaagct catcgcccgc aagaaggact gggatccgaa gaagtacggc
7681 gggttcgaca gccccactgt ggcctactcg gtcctggttg tggcgaaggt tgagaagggc
7741 aagtccaaga agctcaagag cgtgaaggag ctgctgggga tcacgattat ggagcgctcc
7801 agcttcgaga agaacccgat cgatttcctg gaggcgaagg gctacaagga ggtgaagaag
7861 gacctgatca ttaagctccc caagtactca ctcttcgagc tggagaacgg caggaagcgg
7921 atgctggctt ccgctggcga gctgcagaag gggaacgagc tggctctgcc gtccaagtat
7981 gtgaacttcc tctacctggc ctcccactac gagaagctca agggcagccc cgaggacaac
8041 gagcagaagc agctgttcgt cgagcagcac aagcattacc tcgacgagat cattgagcag
8101 atttccgagt tctccaagcg cgtgatcctg gccgacgcga atctggataa ggtcctctcc
8161 gcgtacaaca agcaccgcga caagccaatc agggagcagg ctgagaatat cattcatctc
8221 ttcaccctga cgaacctcgg cgcccctgct gctttcaagt acttcgacac aactatcgat
8281 cgcaagaggt acacaagcac taaggaggtc ctggacgcga ccctcatcca ccagtcgatt
8341 accggcctct acgagacgcg catcgacctg tctcagctcg ggggcgactc cggcggcagc
8401 ccaaagaaga agcggaaggt gtctggaggt tctcctaaga aaaagagaaa agtgtccggc
8461 ggctccccga agaagaagcg caaggtgtag gagctcagag ctttcgttcg tatcatcggt
8521 ttcgacaacg ttcgtcaagt tcaatgcatc agtttcattg cgcacacacc agaatcctac
8581 tgagtttgag tattatggca ttgggaaaac tgtttttctt gtaccatttg ttgtgcttgt
8641 aatttactgt gttttttatt cggttttcgc tatcgaactg tgaaatggaa atggatggag
8701 aagagttaat gaatgatatg gtccttttgt tcattctcaa attaatatta tttgtttttt
8761 ctcttatttg ttgtgtgttg aatttgaaat tataagagat atgcaaacat tttgttttga
8821 gtaaaaatgt gtcaaatcgt ggcctctaat gaccgaagtt aatatgagga gtaaaacact
8881 tgtagttgta ccattatgct tattcactag gcaacaaata tattttcaga cctagaaaag
8941 ctgcaaatgt tactgaatac aagtatgtcc tcttgtgttt tagacattta tgaactttcc
9001 tttatgtaat tttccagaat ccttgtcaga ttctaatcat tgctttataa ttatagttat
9061 actcatggat ttgtagttga gtatgaaaat attttttaat gcattttatg acttgccaat
9121 tgattgacaa cgaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc
9181 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
9241 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
9301 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
9361 gctagagcag cttgccaaca tggtggagca cgacactctc gtctactcca agaatatcaa
9421 agatacagtc tcagaagacc aaagggctat tgagactttt caacaaaggg taatatcggg
9481 aaacctcctc ggattccatt gcccagctat ctgtcacttc atcaaaagga cagtagaaaa
9541 ggaaggtggc acctacaaat gccatcattg cgataaagga aaggctatcg ttcaagatgc
9601 ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga
9661 agacgttcca accacgtctt caaagcaagt ggattgatgt gaacatggtg gagcacgaca
9721 ctctcgtcta ctccaagaat atcaaagata cagtctcaga agaccaaagg gctattgaga
9781 cttttcaaca aagggtaata tcgggaaacc tcctcggatt ccattgccca gctatctgtc
9841 acttcatcaa aaggacagta gaaaaggaag gtggcaccta caaatgccat cattgcgata
9901 aaggaaaggc tatcgttcaa gatgcctctg ccgacagtgg tcccaaagat ggacccccac
9961 ccacgaggag catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt
10021 gatgtgatat ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc
10081 cttcctctat ataaggaagt tcatttcatt tggagaggac acgctgaaat caccagtctc
10141 tctctacaaa tctatctctc tcgagctttc gcagatccgg ggggcaatga gatatgaaaa
10201 agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct
10261 ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag
10321 ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg
10381 tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggagt
10441 ttagcgagag cctgacctat tgcatctccc gccgttcaca gggtgtcacg ttgcaagacc
10501 tgcctgaaac cgaactgccc gctgttctac aaccggtcgc ggaggctatg gatgcgatcg
10561 ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc
10621 aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc
10681 aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc
10741 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca
10801 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg
10861 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg
10921 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccacgactcc
10981 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt
11041 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga
11101 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag
11161 aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aagaaataga
11221 gtagatgccg accgggatct gtcgatcgac aagctcgagt ttctccataa taatgtgtga
11281 gtagttccca gataagggaa ttagggttcc tatagggttt cgctcatgtg ttgagcatat
11341 aagaaaccct tagtatgtat ttgtatttgt aaaatacttc tatcaataaa atttctaatt
11401 cctaaaacca aaatccagta ctaaaatcca gatcccccga attaattcgg cgttaattca
11461 gtacattaaa aacgtccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc
11521 acaatatatc ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc
11581 accactcgat acaggcagcc catcagtccg ggacggcgtc agcgggagag ccgttgtaag
11641 gcggcagact ttgctcatgt taccgatgct attcggaaga acggcaacta agctgccggg
11701 tttgaaacac ggatgatctc gcggagggta gcatgttgat tgtaacgatg acagagcgtt
11761 gctgcctgtg atcaccgcgg tttcaaaatc ggctccgtcg atactatgtt atacgccaac
11821 tttgaaaaca actttgaaaa agctgttttc tggtatttaa ggttttagaa tgcaaggaac
11881 agtgaattgg agttcgtctt gttataatta gcttcttggg gtatctttaa atactgtaga
11941 aaagaggaag gaaataataa atggctaaaa tgagaatatc accggaattg aaaaaactga
12001 tcgaaaaata ccgctgcgta aaagatacgg aaggaatgtc tcctgctaag gtatataagc
12061 tggtgggaga aaatgaaaac ctatatttaa aaatgacgga cagccggtat aaagggacca
12121 cctatgatgt ggaacgggaa aaggacatga tgctatggct ggaaggaaag ctgcctgttc
12181 caaaggtcct gcactttgaa cggcatgatg gctggagcaa tctgctcatg agtgaggccg
12241 atggcgtcct ttgctcggaa gagtatgaag atgaacaaag ccctgaaaag attatcgagc
12301 tgtatgcgga gtgcatcagg ctctttcact ccatcgacat atcggattgt ccctatacga
12361 atagcttaga cagccgctta gccgaattgg attacttact gaataacgat ctggccgatg
12421 tggattgcga aaactgggaa gaagacactc catttaaaga tccgcgcgag ctgtatgatt
12481 ttttaaagac ggaaaagccc gaagaggaac ttgtcttttc ccacggcgac ctgggagaca
12541 gcaacatctt tgtgaaagat ggcaaagtaa gtggctttat tgatcttggg agaagcggca
12601 gggcggacaa gtggtatgac attgccttct gcgtccggtc gatcagggag gatatcgggg
12661 aagaacagta tgtcgagcta ttttttgact tactggggat caagcctgat tgggagaaaa
12721 taaaatatta tattttactg gatgaattgt tttagtacct agaatgcatg accaaaatcc
12781 cttaacgctg atgaatcccc taatgatttt ggtaaaaatc attaagttaa ggtggataca
12841 catcttgtca tatgatcaaa tggtttcgcg aaaaatcaat aatcagacaa caagatgtgc
12901 gaactcgata ttttacacga ctctctttac caattctgcc ccgaattaca cttaaaacga
12961 ctcaacagct taacgttggc ttgccacgca ttacttgact gtaaaactct cactcttacc
13021 gaacttggcc gtaacctgcc aaccaaagcg agaacaaaac ataacatcaa acgaatcgac
13081 cgattgttag gtaatcgtca cctccacaaa gagcgactcg ctgtataccg ttggcatgct
13141 agctttatct gttcgggcaa tacgatgccc attgtacttg ttgactggtc tgatattcgt
13201 gagcaaaaac gacttatggt attgcgagct tcagtcgcac tacacggtcg ttctgttact
13261 ctttatgaga aagcgttccc gctttcagag caatgttcaa agaaagctca tgaccaattt
13321 ctagccgacc ttgcgagcat tctaccgagt aacaccacac cgctcattgt cagtgatgct
13381 ggctttaaag tgccatggta taaatccgtt gagaagctgg gttggtactg gttaagtcga
13441 gtaagaggaa aagtacaata tgcagaccta ggagcggaaa actggaaacc tatcagcaac
13501 ttacatgata tgtcatctag tcactcaaag actttaggct ataagaggct gactaaaagc
13561 aatccaatct catgccaaat tctattgtat aaatctcgct ctaaaggccg aaaaaatcag
13621 cgctcgacac ggactcattg tcaccacccg tcacctaaaa tctactcagc gtcggcaaag
13681 gagccatggg ttctagcaac taacttacct gttgaaattc gaacacccaa acaacttgtt
13741 aatatctatt cgaagcgaat gcagattgaa gaaaccttcc gagacttgaa aagtcctgcc
13801 tacggactag gcctacgcca tagccgaacg agcagctcag agcgttttga tatcatgctg
13861 ctaatcgccc tgatgcttca actaacatgt tggcttgcgg gcgttcatgc tcagaaacaa
13921 ggttgggaca agcacttcca ggctaacaca gtcagaaatc gaaacgtact ctcaacagtt
13981 cgcttaggca tggaagtttt gcggcattct ggctacacaa taacaaggga agacttactc
14041 gtggctgcaa ccctactagc tcaaaattta ttcacacatg gttacgcttt ggggaaatta
14101 tgaggggatc tctcagccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
14161 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
14221 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
14281 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
14341 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
14401 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
14461 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
14521 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
14581 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
14641 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
14701 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
14761 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
14821 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
14881 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
14941 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
15001 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
15061 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
15121 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
15181 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagggtgc
15241 cttgatgtgg gcgccggcgg tcgagtggcg acggcgcggc ttgtccgcgc cctggtagat
15301 tgcctggccg taggccagcc atttttgagc ggccagcggc cgcgataggc cgacgcgaag
15361 cggcggggcg tagggagcgc agcgaccgaa gggtaggcgc tttttgcagc tcttcggctg
15421 tgcgctggcc agacagttat gcacaggcca ggcgggtttt aagagtttta ataagtttta
15481 aagagtttta ggcggaaaaa tcgccttttt tctcttttat atcagtcact tacatgtgtg
15541 accggttccc aatgtacggc tttgggttcc caatgtacgg gttccggttc ccaatgtacg
15601 gctttgggtt cccaatgtac gtgctatcca caggaaacag accttttcga cctttttccc
15661 ctgctagggc aatttgccct agcatctgct ccgtacatta ggaaccggcg gatgcttcgc
15721 cctcgatcag gttgcggtag cgcatgacta ggatcgggcc agcctgcccc gcctcctcct
15781 tcaaatcgta ctccggcagg tcatttgacc cgatcagctt gcgcacggtg aaacagaact
15841 tcttgaactc tccggcgctg ccactgcgtt cgtagatcgt cttgaacaac catctggctt
15901 ctgccttgcc tgcggcgcgg cgtgccaggc ggtagagaaa acggccgatg ccgggatcga
15961 tcaaaaagta atcggggtga accgtcagca cgtccgggtt cttgccttct gtgatctcgc
16021 ggtacatcca atcagctagc tcgatctcga tgtactccgg ccgcccggtt tcgctcttta
16081 cgatcttgta gcggctaatc aaggcttcac cctcggatac cgtcaccagg cggccgttct
16141 tggccttctt cgtacgctgc atggcaacgt gcgtggtgtt taaccgaatg caggtttcta
16201 ccaggtcgtc tttctgcttt ccgccatcgg ctcgccggca gaacttgagt acgtccgcaa
16261 cgtgtggacg gaacacgcgg ccgggcttgt ctcccttccc ttcccggtat cggttcatgg
16321 attcggttag atgggaaacc gccatcagta ccaggtcgta atcccacaca ctggccatgc
16381 cggccggccc tgcggaaacc tctacgtgcc cgtctggaag ctcgtagcgg atcacctcgc
16441 cagctcgtcg gtcacgcttc gacagacgga aaacggccac gtccatgatg ctgcgactat
16501 cgcgggtgcc cacgtcatag agcatcggaa cgaaaaaatc tggttgctcg tcgcccttgg
16561 gcggcttcct aatcgacggc gcaccggctg ccggcggttg ccgggattct ttgcggattc
16621 gatcagcggc cgcttgccac gattcaccgg ggcgtgcttc tgcctcgatg cgttgccgct
16681 gggcggcctg cgcggccttc aacttctcca ccaggtcatc acccagcgcc gcgccgattt
16741 gtaccgggcc ggatggtttg cgaccgctca cgccgattcc tcgggcttgg gggttccagt
16801 gccattgcag ggccggcagg caacccagcc gcttacgcct ggccaaccgc ccgttcctcc
16861 acacatgggg cattccacgg cgtcggtgcc tggttgttct tgattttcca tgccgcctcc
16921 tttagccgct aaaattcatc tactcattta ttcatttgct catttactct ggtagctgcg
16981 cgatgtattc agatagcagc tcggtaatgg tcttgccttg gcgtaccgcg tacatcttca
17041 gcttggtgtg atcctccgcc ggcaactgaa agttgacccg cttcatggct ggcgtgtctg
17101 ccaggctggc caacgttgca gccttgctgc tgcgtgcgct cggacggccg gcacttagcg
17161 tgtttgtgct tttgctcatt ttctctttac ctcattaact caaatgagtt ttgatttaat
17221 ttcagcggcc agcgcctgga cctcgcgggc agcgtcgccc tcgggttctg attcaagaac
17281 ggttgtgccg gcggcggcag tgcctgggta gctcacgcgc tgcgtgatac gggactcaag
17341 aatgggcagc tcgtacccgg ccagcgcctc ggcaacctca ccgccgatgc gcgtgccttt
17401 gatcgcccgc gacacgacaa aggccgcttg tagccttcca tccgtgacct caatgcgctg
17461 cttaaccagc tccaccaggt cggcggtggc ccatatgtcg taagggcttg gctgcaccgg
17521 aatcagcacg aagtcggctg ccttgatcgc ggacacagcc aagtccgccg cctggggcgc
17581 tccgtcgatc actacgaagt cgcgccggcc gatggccttc acgtcgcggt caatcgtcgg
17641 gcggtcgatg ccgacaacgg ttagcggttg atcttcccgc acggccgccc aatcgcgggc
17701 actgccctgg ggatcggaat cgactaacag aacatcggcc ccggcgagtt gcagggcgcg
17761 ggctagatgg gttgcgatgg tcgtcttgcc tgacccgcct ttctggttaa gtacagcgat
17821 aaccttcatg cgttcccctt gcgtatttgt ttatttactc atcgcatcat atacgcagcg
17881 accgcatgac gcaagctgtt ttactcaaat acacatcacc tttttagacg gcggcgctcg
17941 gtttcttcag cggccaagct ggccggccag gccgccagct tggcatcaga caaaccggcc
18001 aggatttcat gcagccgcac ggttgagacg tgcgcgggcg gctcgaacac gtacccggcc
18061 gcgatcatct ccgcctcgat ctcttcggta atgaaaaacg gttcgtcctg gccgtcctgg
18121 tgcggtttca tgcttgttcc tcttggcgtt cattctcggc ggccgccagg gcgtcggcct
18181 cggtcaatgc gtcctcacgg aaggcaccgc gccgcctggc ctcggtgggc gtcacttcct
18241 cgctgcgctc aagtgcgcgg tacagggtcg agcgatgcac gccaagcagt gcagccgcct
18301 ctttcacggt gcggccttcc tggtcgatca gctcgcgggc gtgcgcgatc tgtgccgggg
18361 tgagggtagg gcgggggcca aacttcacgc ctcgggcctt ggcggcctcg cgcccgctcc
18421 gggtgcggtc gatgattagg gaacgctcga actcggcaat gccggcgaac acggtcaaca
18481 ccatgcggcc ggccggcgtg gtggtgtcgg cccacggctc tgccaggcta cgcaggcccg
18541 cgccggcctc ctggatgcgc tcggcaatgt ccagtaggtc gcgggtgctg cgggccaggc
18601 ggtctagcct ggtcactgtc acaacgtcgc cagggcgtag gtggtcaagc atcctggcca
18661 gctccgggcg gtcgcgcctg gtgccggtga tcttctcgga aaacagcttg gtgcagccgg
18721 ccgcgtgcag ttcggcccgt tggttggtca agtcctggtc gtcggtgctg acgcgggcat
18781 agcccagcag gccagcggcg gcgctcttgt tcatggcgta atgtctccgg ttctagtcgc
18841 aagtattcta ctttatgcga ctaaaacac
//
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pH-PABE-7-esgRNA质粒载体应用举例
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