| 质粒名称: | pH-ABE8e-spG |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 200775 |
| 存储实验室: | Caixia Gao |
|
载体骨架基本信息 |
|
| 载体名称: | pHUE411 (modified from pCambia) |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 17211bp |
| 载体类型: | Plant Expression, CRISPR |
| 筛选标记: | Hygromycin |
| 载体抗性: | Kanamycin, 50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | nCas9-spG(D10A)-TadA8e\TadA8e-32aa linker-nCas9-spG(D10A)-3NLS |
| 物种来源: | Streptococcus pyogenes for Cas9; E. coli for TadA |
| 基因突变: | D10A, D1135L, S1136W, G1218K, E1219Q, R1335Q, T1337R for Cas9; W23R, H36L, P48A, R51L, L84F, A106V, D108N, A109S, T111R, D11l9N, H122N, H123Y, S146C, F149Y, R152P, E155V, I156F, K157N, T166I, D167N for TadA |
| 启动子: | maize Ubiquitin-1, OsU3 |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | N/A |
|
文献引用方法 |
|
| 材料和方法部分: | pH-ABE8e-spG was a gift from Caixia Gao (Addgene plasmid # 200775 ; http://n2t.net/addgene:200775 ; RRID:Addgene_200775) |
| 参考文献部分: | Tuning plant phenotypes by precise, graded downregulation of gene expression. Xue C, Qiu F, Wang Y, Li B, Zhao KT, Chen K, Gao C. Nat Biotechnol. 2023 Mar 9. doi: 10.1038/s41587-023-01707-w. 10.1038/s41587-023-01707-w PubMed 36894598 |
买家导航
pH-ABE8e-spG载体质粒基本信息
pH-ABE8e-spG质粒图谱载体图谱和pH-ABE8e-spG载体序列质粒序列多克隆位点信息
pH-ABE8e-spG质粒载体简介
pH-ABE8e-spG质粒序列载体序列
LOCUS pH-ABE8e-spG 17211 bp DNA circular SYN 14-APR-2025
DEFINITION For plant adenine base editing in rice plants or monocotyledons
protoplasts.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 17211)
AUTHORS Xue C, Qiu F, Wang Y, Li B, Zhao KT, Chen K, Gao C
TITLE Tuning plant phenotypes by precise, graded downregulation of gene
expression.
JOURNAL Nat Biotechnol. 2023 Mar 9. doi: 10.1038/s41587-023-01707-w.
PUBMED 36894598
REFERENCE 2 (bases 1 to 17211)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Biotechnol. 2023 Mar 9. doi: 10.1038/s41587-023-01707-w."
COMMENT Sequence Label: pH-ABE8e-spG
FEATURES Location/Qualifiers
source 1..17211
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(578..1207)
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
misc_feature 2508..2532
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
primer_bind 2720..2742
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2734..2751
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2735..2751
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2818..3197
/label=OsU3 promoter
/note="Oryza sativa (rice) snRNA U3 promoter"
misc_RNA 3233..3308
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 3612..5604
/label=Ubi promoter
/note="maize polyubiquitin gene promoter"
CDS 10328..10348
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 10361..10381
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 10394..10414
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(11073..11089)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(11073..11089)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(11086..11108)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 11097..11113
/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(join(11121..11127,11128..11145,11146..11151))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11166..11187
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 11378..12054
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
primer_bind 11991..12011
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
CDS 12121..13146
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
polyA_signal 13187..13361
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
misc_feature 13439..13463
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 13888..14682
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
rep_origin 14769..15357
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 15258..15277
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 15511..15528
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 15543..15683
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(15609..15628)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 15769..15791
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
rep_origin 16027..16221
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(join(16287..17211,1..149))
/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"
ORIGIN
1 tcagcggccg cttgccacga ttcaccgggg cgtgcttctg cctcgatgcg ttgccgctgg
61 gcggcctgcg cggccttcaa cttctccacc aggtcatcac ccagcgccgc gccgatttgt
121 accgggccgg atggtttgcg accgctcacg ccgattcctc gggcttgggg gttccagtgc
181 cattgcaggg ccggcaggca acccagccgc ttacgcctgg ccaaccgccc gttcctccac
241 acatggggca ttccacggcg tcggtgcctg gttgttcttg attttccatg ccgcctcctt
301 tagccgctaa aattcatcta ctcatttatt catttgctca tttactctgg tagctgcgcg
361 atgtattcag atagcagctc ggtaatggtc ttgccttggc gtaccgcgta catcttcagc
421 ttggtgtgat cctccgccgg caactgaaag ttgacccgct tcatggctgg cgtgtctgcc
481 aggctggcca acgttgcagc cttgctgctg cgtgcgctcg gacggccggc acttagcgtg
541 tttgtgcttt tgctcatttt ctctttacct cattaactca aatgagtttt gatttaattt
601 cagcggccag cgcctggacc tcgcgggcag cgtcgccctc gggttctgat tcaagaacgg
661 ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg gactcaagaa
721 tgggcagctc gtacccggcc agcgcctcgg caacctcacc gccgatgcgc gtgcctttga
781 tcgcccgcga cacgacaaag gccgcttgta gccttccatc cgtgacctca atgcgctgct
841 taaccagctc caccaggtcg gcggtggccc atatgtcgta agggcttggc tgcaccggaa
901 tcagcacgaa gtcggctgcc ttgatcgcgg acacagccaa gtccgccgcc tggggcgctc
961 cgtcgatcac tacgaagtcg cgccggccga tggccttcac gtcgcggtca atcgtcgggc
1021 ggtcgatgcc gacaacggtt agcggttgat cttcccgcac ggccgcccaa tcgcgggcac
1081 tgccctgggg atcggaatcg actaacagaa catcggcccc ggcgagttgc agggcgcggg
1141 ctagatgggt tgcgatggtc gtcttgcctg acccgccttt ctggttaagt acagcgataa
1201 ccttcatgcg ttccccttgc gtatttgttt atttactcat cgcatcatat acgcagcgac
1261 cgcatgacgc aagctgtttt actcaaatac acatcacctt tttagacggc ggcgctcggt
1321 ttcttcagcg gccaagctgg ccggccaggc cgccagcttg gcatcagaca aaccggccag
1381 gatttcatgc agccgcacgg ttgagacgtg cgcgggcggc tcgaacacgt acccggccgc
1441 gatcatctcc gcctcgatct cttcggtaat gaaaaacggt tcgtcctggc cgtcctggtg
1501 cggtttcatg cttgttcctc ttggcgttca ttctcggcgg ccgccagggc gtcggcctcg
1561 gtcaatgcgt cctcacggaa ggcaccgcgc cgcctggcct cggtgggcgt cacttcctcg
1621 ctgcgctcaa gtgcgcggta cagggtcgag cgatgcacgc caagcagtgc agccgcctct
1681 ttcacggtgc ggccttcctg gtcgatcagc tcgcgggcgt gcgcgatctg tgccggggtg
1741 agggtagggc gggggccaaa cttcacgcct cgggccttgg cggcctcgcg cccgctccgg
1801 gtgcggtcga tgattaggga acgctcgaac tcggcaatgc cggcgaacac ggtcaacacc
1861 atgcggccgg ccggcgtggt ggtgtcggcc cacggctctg ccaggctacg caggcccgcg
1921 ccggcctcct ggatgcgctc ggcaatgtcc agtaggtcgc gggtgctgcg ggccaggcgg
1981 tctagcctgg tcactgtcac aacgtcgcca gggcgtaggt ggtcaagcat cctggccagc
2041 tccgggcggt cgcgcctggt gccggtgatc ttctcggaaa acagcttggt gcagccggcc
2101 gcgtgcagtt cggcccgttg gttggtcaag tcctggtcgt cggtgctgac gcgggcatag
2161 cccagcaggc cagcggcggc gctcttgttc atggcgtaat gtctccggtt ctagtcgcaa
2221 gtattctact ttatgcgact aaaacacgcg acaagaaaac gccaggaaaa gggcagggcg
2281 gcagcctgtc gcgtaactta ggacttgtgc gacatgtcgt tttcagaaga cggctgcact
2341 gaacgtcaga agccgactgc actatagcag cggaggggtt ggatcaaagt actttgatcc
2401 cgaggggaac cctgtggttg gcatgcacat acaaatggac gaacggataa accttttcac
2461 gcccttttaa atatccgatt attctaataa acgctctttt ctcttaggtt tacccgccaa
2521 tatatcctgt caaacactga tagtttaaac tgaaggcggg aaacgacaat ctgatccaag
2581 ctcaagctgc tctagcattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
2641 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
2701 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctt
2761 agtaattcat ccaggtctcc aagttctagg attttcagaa ctgcaactta ttttatcaag
2821 gaatctttaa acatacgaac agatcactta aagttcttct gaagcaactt aaagttatca
2881 ggcatgcatg gatcttggag gaatcagatg tgcagtcagg gaccatagca caagacaggc
2941 gtcttctact ggtgctacca gcaaatgctg gaagccggga acactgggta cgttggaaac
3001 cacgtgatgt gaagaagtaa gataaactgt aggagaaaag catttcgtag tgggccatga
3061 agcctttcag gacatgtatt gcagtatggg ccggcccatt acgcaattgg acgacaacaa
3121 agactagtat tagtaccacc tcggctatcc acatagatca aagctgattt aaaagagttg
3181 tgcagatgat ccgtggcgaa ccgcaggtga gctgcattaa tcacctgcgc aagttttaga
3241 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
3301 gtcggtgctt ttttttttcg ttttgcattg agttttctcc gtcgcatgtt tgcagtttta
3361 ttttccgttt tgcattgaaa tttctccgtc tcatgtttgc agcgtgttca aaaagtacgc
3421 agctgtattt cacttattta cggcgccaca ttttcatgcc gtttgtgcca actatcccga
3481 gctagtgaat acagcttggc ttcacacaac actggtgacc cgctgacctg ctcgtacctc
3541 gtaccgtcgt acggcacagc atttggaatt aaagggtgtg atcgatactg cttgctgcta
3601 agcttgcatg cctgcagtgc agcgtgaccc ggtcgtgccc ctctctagag ataatgagca
3661 ttgcatgtct aagttataaa aaattaccac atattttttt tgtcacactt gtttgaagtg
3721 cagtttatct atctttatac atatatttaa actttactct acgaataata taatctatag
3781 tactacaata atatcagtgt tttagagaat catataaatg aacagttaga catggtctaa
3841 aggacaattg agtattttga caacaggact ctacagtttt atctttttag tgtgcatgtg
3901 ttctcctttt tttttgcaaa tagcttcacc tatataatac ttcatccatt ttattagtac
3961 atccatttag ggtttagggt taatggtttt tatagactaa tttttttagt acatctattt
4021 tattctattt tagcctctaa attaagaaaa ctaaaactct attttagttt ttttatttaa
4081 taatttagat ataaaataga ataaaataaa gtgactaaaa attaaacaaa taccctttaa
4141 gaaattaaaa aaactaagga aacatttttc ttgtttcgag tagataatgc cagcctgtta
4201 aacgccgtcg acgagtctaa cggacaccaa ccagcgaacc agcagcgtcg cgtcgggcca
4261 agcgaagcag acggcacggc atctctgtcg ctgcctctgg acccctctcg agagttccgc
4321 tccaccgttg gacttgctcc gctgtcggca tccagaaatt gcgtggcgga gcggcagacg
4381 tgagccggca cggcaggcgg cctcctcctc ctctcacggc accggcagct acgggggatt
4441 cctttcccac cgctccttcg ctttcccttc ctcgcccgcc gtaataaata gacaccccct
4501 ccacaccctc tttccccaac ctcgtgttgt tcggagcgca cacacacaca accagatctc
4561 ccccaaatcc acccgtcggc acctccgctt caaggtacgc cgctcgtcct cccccccccc
4621 ccctctctac cttctctaga tcggcgttcc ggtccatggt tagggcccgg tagttctact
4681 tctgttcatg tttgtgttag atccgtgttt gtgttagatc cgtgctgcta gcgttcgtac
4741 acggatgcga cctgtacgtc agacacgttc tgattgctaa cttgccagtg tttctctttg
4801 gggaatcctg ggatggctct agccgttccg cagacgggat cgatttcatg attttttttg
4861 tttcgttgca tagggtttgg tttgcccttt tcctttattt caatatatgc cgtgcacttg
4921 tttgtcgggt catcttttca tgcttttttt tgtcttggtt gtgatgatgt ggtctggttg
4981 ggcggtcgtt ctagatcgga gtagaattct gtttcaaact acctggtgga tttattaatt
5041 ttggatctgt atgtgtgtgc catacatatt catagttacg aattgaagat gatggatgga
5101 aatatcgatc taggataggt atacatgttg atgcgggttt tactgatgca tatacagaga
5161 tgctttttgt tcgcttggtt gtgatgatgt ggtgtggttg ggcggtcgtt cattcgttct
5221 agatcggagt agaatactgt ttcaaactac ctggtgtatt tattaatttt ggaactgtat
5281 gtgtgtgtca tacatcttca tagttacgag tttaagatgg atggaaatat cgatctagga
5341 taggtataca tgttgatgtg ggttttactg atgcatatac atgatggcat atgcagcatc
5401 tattcatatg ctctaacctt gagtacctat ctattataat aaacaagtat gttttataat
5461 tattttgatc ttgatatact tggatgatgg catatgcagc agctatatgt ggattttttt
5521 agccctgcct tcatacgcta tttatttgct tggtactgtt tcttttgtcg atgctcaccc
5581 tgttgtttgg tgttacttct gcagccctag gatgagcgaa gttgaattta gccatgaata
5641 ttggatgcgc catgcactta cacttgcaaa gagagcccgc gatgaacgcg aagttcctgt
5701 tggagcagtt cttgttctta acaaccgcgt tatcggagaa ggatggaacc gcgcaatcgg
5761 acttcatgat cctacagcac atgcagaaat catggcactt cgccagggag gacttgttat
5821 gcagaactat cgccttatcg atgcaacact ttatgttaca tttgaacctt gcgttatgtg
5881 cgcaggagca atgatccata gccgcatcgg acgcgttgtg ttcggagtac gtaacagcaa
5941 acgcggagca gcaggaagcc ttatgaacgt tcttaactat cctggaatga accatcgcgt
6001 tgaaatcaca gaaggaatcc ttgcagatga atgcgcagca cttctttgcg atttctaccg
6061 tatgcctcgc caggtgttca atgcgcagaa gaaggcccaa agcagcatca actcaggtgg
6121 tagctccggc ggctcgtcag gttccgaaac acctggaaca agcgaaagcg caacacctga
6181 atctagtgga ggaagttctg gtgggtccct taaggacaag aagtactcga tcggcctcgc
6241 catcgggacg aactcagttg gctgggccgt gatcaccgac gagtacaagg tgccctctaa
6301 gaagttcaag gtcctgggga acaccgaccg ccattccatc aagaagaacc tcatcggcgc
6361 tctcctgttc gacagcgggg agaccgctga ggctacgagg ctcaagagaa ccgctaggcg
6421 ccggtacacg agaaggaaga acaggatctg ctacctccaa gagattttct ccaacgagat
6481 ggccaaggtt gacgattcat tcttccaccg cctggaggag tctttcctcg tggaggagga
6541 taagaagcac gagcggcatc ccatcttcgg caacatcgtg gacgaggttg cctaccacga
6601 gaagtaccct acgatctacc atctgcggaa gaagctcgtg gactccaccg ataaggcgga
6661 cctcagactg atctacctcg ctctggccca catgatcaag ttccgcggcc atttcctgat
6721 cgagggggat ctcaacccag acaacagcga tgttgacaag ctgttcatcc aactcgtgca
6781 gacctacaac caactcttcg aggagaaccc gatcaacgcc tctggcgtgg acgcgaaggc
6841 tatcctgtcc gcgaggctct cgaagtccag gaggctggag aacctgatcg ctcagctccc
6901 aggcgagaag aagaacggcc tgttcgggaa cctcatcgct ctcagcctgg ggctcacccc
6961 gaacttcaag tcgaacttcg atctcgctga ggacgccaag ctgcaactct ccaaggacac
7021 ctacgacgat gacctcgata acctcctggc ccagatcggc gatcaatacg cggacctgtt
7081 cctcgctgcc aagaacctgt cggacgccat cctcctgtca gatatcctcc gcgtgaacac
7141 cgagatcacg aaggctccac tctctgcctc catgatcaag cgctacgacg agcaccatca
7201 ggatctgacc ctcctgaagg cgctggtccg ccaacagctc ccggagaagt acaaggagat
7261 tttcttcgat cagtcgaaga acggctacgc tgggtacatc gacggcgggg cctcacaaga
7321 ggagttctac aagttcatca agccaatcct ggagaagatg gacggcacgg aggagctcct
7381 ggtgaagctc aacagggagg acctcctgcg gaagcagaga accttcgata acggcagcat
7441 cccccaccaa atccatctcg gggagctgca cgccatcctg agaaggcaag aggacttcta
7501 ccctttcctc aaggataacc gggagaagat cgagaagatc ctgaccttca gaatcccata
7561 ctacgtcggc cctctcgcgc gggggaactc aagattcgct tggatgaccc gcaagtctga
7621 ggagaccatc acgccgtgga acttcgagga ggtggtggac aagggcgcta gcgctcagtc
7681 gttcatcgag aggatgacca acttcgacaa gaacctgccc aacgagaagg tgctccctaa
7741 gcactcgctc ctgtacgagt acttcaccgt ctacaacgag ctcacgaagg tgaagtacgt
7801 caccgagggc atgcgcaagc cagcgttcct gtccggggag cagaagaagg ctatcgtgga
7861 cctcctgttc aagaccaacc ggaaggtcac ggttaagcaa ctcaaggagg actacttcaa
7921 gaagatcgag tgcttcgatt cggtcgagat cagcggcgtt gaggaccgct tcaacgccag
7981 cctcgggacc taccacgatc tcctgaagat catcaaggat aaggacttcc tggacaacga
8041 ggagaacgag gatatcctgg aggacatcgt gctgaccctc acgctgttcg aggacaggga
8101 gatgatcgag gagcgcctga agacgtacgc ccatctcttc gatgacaagg tcatgaagca
8161 actcaagcgc cggagataca ccggctgggg gaggctgtcc cgcaagctca tcaacggcat
8221 ccgggacaag cagtccggga agaccatcct cgacttcctc aagagcgatg gcttcgccaa
8281 caggaacttc atgcaactga tccacgatga cagcctcacc ttcaaggagg atatccaaaa
8341 ggctcaagtg agcggccagg gggactcgct gcacgagcat atcgcgaacc tcgctggctc
8401 ccccgcgatc aagaagggca tcctccagac cgtgaaggtt gtggacgagc tcgtgaaggt
8461 catgggccgg cacaagcctg agaacatcgt catcgagatg gccagagaga accaaaccac
8521 gcagaagggg caaaagaact ctagggagcg catgaagcgc atcgaggagg gcatcaagga
8581 gctggggtcc caaatcctca aggagcaccc agtggagaac acccaactgc agaacgagaa
8641 gctctacctg tactacctcc agaacggcag ggatatgtac gtggaccaag agctggatat
8701 caaccgcctc agcgattacg acgtcgatca tatcgttccc cagtctttcc tgaaggatga
8761 ctccatcgac aacaaggtcc tcaccaggtc ggacaagaac cgcggcaagt cagataacgt
8821 tccatctgag gaggtcgtta agaagatgaa gaactactgg aggcagctcc tgaacgccaa
8881 gctgatcacg caaaggaagt tcgacaacct caccaaggct gagagaggcg ggctctcaga
8941 gctggacaag gccggcttca tcaagcggca gctggtcgag accagacaaa tcacgaagca
9001 cgttgcgcaa atcctcgact ctcggatgaa cacgaagtac gatgagaacg acaagctgat
9061 cagggaggtt aaggtgatca ccctgaagtc taagctcgtc tccgacttca ggaaggattt
9121 ccagttctac aaggttcgcg agatcaacaa ctaccaccat gcccatgacg cttacctcaa
9181 cgctgtggtc ggcaccgctc tgatcaagaa gtacccaaag ctggagtccg agttcgtgta
9241 cggggactac aaggtttacg atgtgcgcaa gatgatcgcc aagtcggagc aagagatcgg
9301 caaggctacc gccaagtact tcttctactc aaacatcatg aacttcttca agaccgagat
9361 cacgctggcc aacggcgaga tccggaagag accgctcatc gagaccaacg gcgagacggg
9421 ggagatcgtg tgggacaagg gcagggattt cgcgaccgtc cgcaaggttc tctccatgcc
9481 ccaggtgaac atcgtcaaga agaccgaggt ccaaacgggc gggttctcaa aggagtctat
9541 cctgcctaag cggaacagcg acaagctcat cgccagaaag aaggactggg acccaaagaa
9601 gtacggcggg ttcctctggc ctaccgtggc ctactcggtc ctggttgtgg cgaaggttga
9661 gaagggcaag tccaagaagc tcaagagcgt gaaggagctc ctggggatca ccatcatgga
9721 gaggtccagc ttcgagaaga acccaatcga cttcctggag gccaagggct acaaggaggt
9781 gaagaaggac ctgatcatca agctcccgaa gtactctctc ttcgagctgg agaacggcag
9841 gaagagaatg ctggcttccg ctaagcagct ccagaagggg aacgagctcg cgctgccaag
9901 caagtacgtg aacttcctct acctggcttc ccactacgag aagctcaagg gcagcccgga
9961 ggacaacgag caaaagcagc tgttcgtcga gcagcacaag cattacctcg acgagatcat
10021 cgagcaaatc tccgagttca gcaagcgcgt gatcctcgcc gacgcgaacc tggataaggt
10081 cctctccgcc tacaacaagc accgggacaa gcccatcaga gagcaagcgg agaacatcat
10141 ccatctcttc accctgacga acctcggcgc tcctgctgct ttcaagtact tcgacaccac
10201 gatcgatcgg aagcaataca ggtccacgaa ggaggtcctg gacgcgaccc tcatccacca
10261 gtcgatcacc ggcctgtacg agacgaggat cgacctctca caactcggcg gggattccgg
10321 cggcagccca aagaagaagc ggaaggtgtc tggaggttct cctaagaaaa agagaaaagt
10381 gtccggcggc tccccgaaga agaagcgcaa ggtgtaggag ctcagagctt tcgttcgtat
10441 catcggtttc gacaacgttc gtcaagttca atgcatcagt ttcattgcgc acacaccaga
10501 atcctactga gtttgagtat tatggcattg ggaaaactgt ttttcttgta ccatttgttg
10561 tgcttgtaat ttactgtgtt ttttattcgg ttttcgctat cgaactgtga aatggaaatg
10621 gatggagaag agttaatgaa tgatatggtc cttttgttca ttctcaaatt aatattattt
10681 gttttttctc ttatttgttg tgtgttgaat ttgaaattat aagagatatg caaacatttt
10741 gttttgagta aaaatgtgtc aaatcgtggc ctctaatgac cgaagttaat atgaggagta
10801 aaacacttgt agttgtacca ttatgcttat tcactaggca acaaatatat tttcagacct
10861 agaaaagctg caaatgttac tgaatacaag tatgtcctct tgtgttttag acatttatga
10921 actttccttt atgtaatttt ccagaatcct tgtcagattc taatcattgc tttataatta
10981 tagttatact catggatttg tagttgagta tgaaaatatt ttttaatgca ttttatgact
11041 tgccaattga ttgacaacga attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt
11101 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
11161 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
11221 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
11281 cgtattggct agagcagctt gccaacatgg tggagcacga cactctcgtc tactccaaga
11341 atatcaaaga tacagtctca gaagaccaaa gggctattga gacttttcaa caaagggtaa
11401 tatcgggaaa cctcctcgga ttccattgcc cagctatctg tcacttcatc aaaaggacag
11461 tagaaaagga aggtggcacc tacaaatgcc atcattgcga taaaggaaag gctatcgttc
11521 aagatgcctc tgccgacagt ggtcccaaag atggaccccc acccacgagg agcatcgtgg
11581 aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga ttgatgtgaa catggtggag
11641 cacgacactc tcgtctactc caagaatatc aaagatacag tctcagaaga ccaaagggct
11701 attgagactt ttcaacaaag ggtaatatcg ggaaacctcc tcggattcca ttgcccagct
11761 atctgtcact tcatcaaaag gacagtagaa aaggaaggtg gcacctacaa atgccatcat
11821 tgcgataaag gaaaggctat cgttcaagat gcctctgccg acagtggtcc caaagatgga
11881 cccccaccca cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa
11941 gtggattgat gtgatatctc cactgacgta agggatgacg cacaatccca ctatccttcg
12001 caagaccctt cctctatata aggaagttca tttcatttgg agaggacacg ctgaaatcac
12061 cagtctctct ctacaaatct atctctctcg agctttcgca gatccggggg gcaatgagat
12121 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
12181 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
12241 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
12301 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
12361 ggggagttta gcgagagcct gacctattgc atctcccgcc gttcacaggg tgtcacgttg
12421 caagacctgc ctgaaaccga actgcccgct gttctacaac cggtcgcgga ggctatggat
12481 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
12541 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
12601 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
12661 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
12721 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
12781 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
12841 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgcca
12901 cgactccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
12961 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
13021 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
13081 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
13141 aaatagagta gatgccgacc gggatctgtc gatcgacaag ctcgagtttc tccataataa
13201 tgtgtgagta gttcccagat aagggaatta gggttcctat agggtttcgc tcatgtgttg
13261 agcatataag aaacccttag tatgtatttg tatttgtaaa atacttctat caataaaatt
13321 tctaattcct aaaaccaaaa tccagtacta aaatccagat cccccgaatt aattcggcgt
13381 taattcagta cattaaaaac gtccgcaatg tgttattaag ttgtctaagc gtcaatttgt
13441 ttacaccaca atatatcctg ccaccagcca gccaacagct ccccgaccgg cagctcggca
13501 caaaatcacc actcgataca ggcagcccat cagtccggga cggcgtcagc gggagagccg
13561 ttgtaaggcg gcagactttg ctcatgttac cgatgctatt cggaagaacg gcaactaagc
13621 tgccgggttt gaaacacgga tgatctcgcg gagggtagca tgttgattgt aacgatgaca
13681 gagcgttgct gcctgtgatc accgcggttt caaaatcggc tccgtcgata ctatgttata
13741 cgccaacttt gaaaacaact ttgaaaaagc tgttttctgg tatttaaggt tttagaatgc
13801 aaggaacagt gaattggagt tcgtcttgtt ataattagct tcttggggta tctttaaata
13861 ctgtagaaaa gaggaaggaa ataataaatg gctaaaatga gaatatcacc ggaattgaaa
13921 aaactgatcg aaaaataccg ctgcgtaaaa gatacggaag gaatgtctcc tgctaaggta
13981 tataagctgg tgggagaaaa tgaaaaccta tatttaaaaa tgacggacag ccggtataaa
14041 gggaccacct atgatgtgga acgggaaaag gacatgatgc tatggctgga aggaaagctg
14101 cctgttccaa aggtcctgca ctttgaacgg catgatggct ggagcaatct gctcatgagt
14161 gaggccgatg gcgtcctttg ctcggaagag tatgaagatg aacaaagccc tgaaaagatt
14221 atcgagctgt atgcggagtg catcaggctc tttcactcca tcgacatatc ggattgtccc
14281 tatacgaata gcttagacag ccgcttagcc gaattggatt acttactgaa taacgatctg
14341 gccgatgtgg attgcgaaaa ctgggaagaa gacactccat ttaaagatcc gcgcgagctg
14401 tatgattttt taaagacgga aaagcccgaa gaggaacttg tcttttccca cggcgacctg
14461 ggagacagca acatctttgt gaaagatggc aaagtaagtg gctttattga tcttgggaga
14521 agcggcaggg cggacaagtg gtatgacatt gccttctgcg tccggtcgat cagggaggat
14581 atcggggaag aacagtatgt cgagctattt tttgacttac tggggatcaa gcctgattgg
14641 gagaaaataa aatattatat tttactggat gaattgtttt agtacctaga atgcatgacc
14701 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
14761 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
14821 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
14881 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
14941 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
15001 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
15061 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
15121 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
15181 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
15241 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
15301 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
15361 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
15421 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
15481 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
15541 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
15601 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
15661 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
15721 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
15781 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagggtgcc ttgatgtggg
15841 cgccggcggt cgagtggcga cggcgcggct tgtccgcgcc ctggtagatt gcctggccgt
15901 aggccagcca tttttgagcg gccagcggcc gcgataggcc gacgcgaagc ggcggggcgt
15961 agggagcgca gcgaccgaag ggtaggcgct ttttgcagct cttcggctgt gcgctggcca
16021 gacagttatg cacaggccag gcgggtttta agagttttaa taagttttaa agagttttag
16081 gcggaaaaat cgcctttttt ctcttttata tcagtcactt acatgtgtga ccggttccca
16141 atgtacggct ttgggttccc aatgtacggg ttccggttcc caatgtacgg ctttgggttc
16201 ccaatgtacg tgctatccac aggaaacaga ccttttcgac ctttttcccc tgctagggca
16261 atttgcccta gcatctgctc cgtacattag gaaccggcgg atgcttcgcc ctcgatcagg
16321 ttgcggtagc gcatgactag gatcgggcca gcctgccccg cctcctcctt caaatcgtac
16381 tccggcaggt catttgaccc gatcagcttg cgcacggtga aacagaactt cttgaactct
16441 ccggcgctgc cactgcgttc gtagatcgtc ttgaacaacc atctggcttc tgccttgcct
16501 gcggcgcggc gtgccaggcg gtagagaaaa cggccgatgc cgggatcgat caaaaagtaa
16561 tcggggtgaa ccgtcagcac gtccgggttc ttgccttctg tgatctcgcg gtacatccaa
16621 tcagctagct cgatctcgat gtactccggc cgcccggttt cgctctttac gatcttgtag
16681 cggctaatca aggcttcacc ctcggatacc gtcaccaggc ggccgttctt ggccttcttc
16741 gtacgctgca tggcaacgtg cgtggtgttt aaccgaatgc aggtttctac caggtcgtct
16801 ttctgctttc cgccatcggc tcgccggcag aacttgagta cgtccgcaac gtgtggacgg
16861 aacacgcggc cgggcttgtc tcccttccct tcccggtatc ggttcatgga ttcggttaga
16921 tgggaaaccg ccatcagtac caggtcgtaa tcccacacac tggccatgcc ggccggccct
16981 gcggaaacct ctacgtgccc gtctggaagc tcgtagcgga tcacctcgcc agctcgtcgg
17041 tcacgcttcg acagacggaa aacggccacg tccatgatgc tgcgactatc gcgggtgccc
17101 acgtcataga gcatcggaac gaaaaaatct ggttgctcgt cgcccttggg cggcttccta
17161 atcgacggcg caccggctgc cggcggttgc cgggattctt tgcggattcg a
//
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pH-ABE8e-spG质粒载体应用举例
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