質粒名稱: | pJK1 |
---|---|
出品公司: | Addgene |
目錄編號: | 111722 |
存儲實驗室: | Ian Hickson |
載體骨架基本信息 |
|
載體名稱: | pYES2 |
載體出品公司: | ThermoFisher |
完整質粒大小: | 10092bp |
載體類型: | Yeast Expression |
篩選標記: | URA3 |
載體抗性: | 氨芐霉素 |
生長溫度: | 37 ℃ |
宿主菌: | DH5a |
拷貝數: | 高拷貝 |
插入基因信息 |
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插入基因名稱: | BLM、BS、BS |
物種來源: | H. sapiens (human) |
基因突變: | C-terminal hexahistidine tag was added, a yeast Kozak consensus sequence was included upstream of the coding sequence, and the first five codons of S. cerevisiae TOP2 gene replaced the initiation codon of the BLM cDNA. |
啟動子: | GAL1 |
標簽/融合蛋白: | hexahistidine (C terminal on insert) |
克隆方法: | 限制性內切酶 |
5’克隆位點: | known (unknown if destroyed) |
3’克隆位點: | known (unknown if destroyed) |
5’測序引物 | unknown |
文獻引用方法 |
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材料和方法部分: | pJK1 was a gift from Ian Hickson (Addgene plasmid # 111722 ; //n2t.net/addgene:111722 ; RRID:Addgene_111722) |
參考文獻部分: | The Bloom's syndrome gene product is a 3'-5' DNA helicase. Karow JK, Chakraverty RK, Hickson ID. J Biol Chem. 1997 Dec 5;272(49):30611-4. PubMed 9388193 |
買家導航
pJK1載體質粒基本信息
pJK1質粒圖譜載體圖譜和pJK1載體序列質粒序列多克隆位點信息
pJK1質粒載體簡介
pJK1質粒序列載體序列
LOCUS pJK1 10092 bp DNA circular SYN 14-APR-2025 DEFINITION to express human BLM protein in yeast cells. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10092) AUTHORS Karow JK, Chakraverty RK, Hickson ID TITLE The Bloom's syndrome gene product is a 3'-5' DNA helicase. JOURNAL J Biol Chem. 1997 Dec 5;272(49):30611-4. PUBMED 9388193 REFERENCE 2 (bases 1 to 10092) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1 //gt12365.cn COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol Chem."; date: "1997-12-5"; volume: "272(49)"; pages: "30611-4" COMMENT Sequence Label: pJK1 FEATURES Location/Qualifiers source 1..10092 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 1..442 /gene="S. cerevisiae GAL1" /label=GAL1 promoter /note="inducible promoter, regulated by Gal4" protein_bind 1..118 /label=UAS /bound_moiety="Gal4" /note="upstream activating sequence mediating Gal4-dependent induction" primer_bind 413..436 /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" primer_bind 474..493 /label=T7 /note="T7 promoter, forward primer" promoter 474..492 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 4793..4810 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 4840..5087 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(4856..4874) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind complement(5164..5181) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5335..5923) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5415..5434) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(join(6094..6885,6886..6954)) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6717..6736 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" CDS complement(7050..7853) /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /label=URA3 /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" promoter complement(7854..8069) /gene="S. cerevisiae URA3" /label=URA3 promoter rep_origin 8670..9547 /label=2u ori /note="yeast 2u plasmid origin of replication" rep_origin complement(9558..10071) /direction=LEFT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(9753..9774) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 9965..9984 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 cggattagaa gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc 61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa 121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc 181 tggccccaca aaccttcaaa tgaacgaatc aaattaacaa ccataggatg ataatgcgat 241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt 301 aacagatata taaatgcaaa aactgcataa ccactttaac taatactttc aacattttcg 361 gtttgtatta cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc 421 tctatacttt aacgtcaagg agaaaaaacc ccggatcgga ctactagcag ctgtaatacg 481 actcactata gggaatatta agcttggtac cgagctcgga tccctaacca tgtcaactga 541 accggctgct gttcctcaaa ataatctaca ggagcaacta gaacgtcact cagccagaac 601 acttaataat aaattaagtc tttcaaaacc aaaattttca ggtttcactt ttaaaaagaa 661 aacatcttca gataacaatg tatctgtaac taatgtgtca gtagcaaaaa cacctgtatt 721 aagaaataaa gatgttaatg ttaccgaaga cttttccttc agtgaacctc tacccaacac 781 cacgaatcag caaagggtca aggacttctt taaaaatgct ccagcaggac aggaaacaca 841 gagaggtgga tcaaaatcat tattgccaga tttcttgcag actccgaagg aagttgtatg 901 cactacccaa aacacaccaa ctgtaaagaa atcccgggat actgctctca agaaattaga 961 atttagttct tcaccagatt ctttaagtac catcaatgat tgggatgata tggatgactt 1021 tgatacttct gagacttcaa aatcatttgt tacaccaccc caaagtcact ttgtaagagt 1081 aagcactgct cagaaatcaa aaaagggtaa gagaaacttt tttaaagcac agctttatac 1141 aacaaacaca gtaaagactg acttgcctcc accctcctct gaaagcgagc aaatagattt 1201 gactgaggaa cagaaggatg actcagaatg gttaagcagc gatgtgattt gcatcgatga 1261 tggccccatt gctgaagtgc atataaatga agatgctcag gaaagtgact ctctgaaaac 1321 tcatttggaa gatgaaagag ataatagcga aaagaagaag aatttggaag aagctgaatt 1381 acattcaact gagaaagttc catgtattga atttgatgat gatgattatg atacggattt 1441 tgttccacct tctccagaag aaattatttc tgcttcttct tcctcttcaa aatgccttag 1501 tacgttaaag gaccttgaca cctctgacag aaaagaggat gttcttagca catcaaaaga 1561 tcttttgtca aaacctgaga aaatgagtat gcaggagctg aatccagaaa ccagcacaga 1621 ctgtgacgct agacagataa gtttacagca gcagcttatt catgtgatgg agcacatctg 1681 taaattaatt gatactattc ctgatgataa actgaaactt ttggattgtg ggaacgaact 1741 gcttcagcag cggaacataa gaaggaaact tctaacggaa gtagatttta ataaaagtga 1801 tgccagtctt cttggctcat tgtggagata caggcctgat tcacttgatg gccctatgga 1861 gggtgattcc tgccctacag ggaattctat gaaggagtta aatttttcac accttccctc 1921 aaattctgtt tctcctgggg actgtttact gactaccacc ctaggaaaga caggattctc 1981 tgccaccagg aagaatcttt ttgaaaggcc tttattcaat acccatttac agaagtcctt 2041 tgtaagtagc aactgggctg aaacaccaag actaggaaaa aaaaatgaaa gctcttattt 2101 cccaggaaat gttctcacaa gcactgctgt gaaagatcag aataaacata ctgcttcaat 2161 aaatgactta gaaagagaaa cccaaccttc ctatgatatt gataattttg acatagatga 2221 ctttgatgat gatgatgact gggaagacat aatgcataat ttagcagcca gcaaatcttc 2281 cacagctgcc tatcaaccca tcaaggaagg tcggccaatt aaatcagtat cagaaagact 2341 ttcctcagcc aagacagact gtcttccagt gtcatctact gctcaaaata taaacttctc 2401 agagtcaatt cagaattata ctgacaagtc agcacaaaat ttagcatcca gaaatctgaa 2461 acatgagcgt ttccaaagtc ttagttttcc tcatacaaag gaaatgatga agatttttca 2521 taaaaaattt ggcctgcata attttagaac taatcagcta gaggcgatca atgctgcact 2581 gcttggtgaa gactgtttta tcctgatgcc gactggaggt ggtaagagtt tgtgttacca 2641 gctccctgcc tgtgtttctc ctggggtcac tgttgtcatt tctcccttga gatcacttat 2701 cgtagatcaa gtccaaaagc tgacttcctt ggatattcca gctacatatc tgacaggtga 2761 taagactgac tcagaagcta caaatattta cctccagtta tcaaaaaaag acccaatcat 2821 aaaactccta tatgtcactc cagaaaagat ctgtgcaagt aacagactca tttctactct 2881 ggagaatctc tatgagagga agctcttggc acgttttgtt attgatgaag cacattgtgt 2941 cagtcagtgg ggacatgatt ttcgtcaaga ttacaaaaga atgaatatgc ttcgccagaa 3001 gtttccttct gttccggtga tggctcttac ggccacagct aatcccaggg tacagaagga 3061 catcctgact cagctgaaga ttctcagacc tcaggtgttt agcatgagct ttaacagaca 3121 taatctgaaa tactatgtat taccgaaaaa gcctaaaaag gtggcatttg attgcctaga 3181 atggatcaga aagcaccacc catatgattc agggataatt tactgcctct ccaggcgaga 3241 atgtgacacc atggctgaca cgttacagag agatgggctc gctgctcttg cttaccatgc 3301 tggcctcagt gattctgcca gagatgaagt gcagcagaag tggattaatc aggatggctg 3361 tcaggttatc tgtgctacaa ttgcatttgg aatggggatt gacaaaccgg acgtgcgatt 3421 tgtgattcat gcatctctcc ctaaatctgt ggagggttac taccaagaat ctggcagagc 3481 tggaagagat ggggaaatat ctcactgcct gcttttctat acctatcatg atgtgaccag 3541 actgaaaaga cttataatga tggaaaaaga tggaaaccat catacaagag aaactcactt 3601 caataatttg tatagcatgg tacattactg tgaaaatata acagaatgca ggagaataca 3661 gcttttggcc tactttggtg aaaatggatt taatcctgat ttttgtaaga aacacccaga 3721 tgtttcttgt gataattgct gtaaaacaaa ggattataaa acaagagatg tgactgacga 3781 tgtgaaaagt attgtaagat ttgttcaaga acatagttca tcacaaggaa tgagaaatat 3841 aaaacatgta ggtccttctg gaagatttac tatgaatatg ctggtcgaca ttttcttggg 3901 gagtaagagt gcaaaaatcc agtcaggtat atttggaaaa ggatctgctt attcacgaca 3961 caatgccgaa agacttttta aaaagctgat acttgacaag attttggatg aagacttata 4021 tatcaatgcc aatgaccagg cgatcgctta tgtgatgctc ggaaataaag cacaaactgt 4081 actaaatggc aatttaaagg tagactttat ggaaacagaa aattccagca gtgtgaaaaa 4141 acaaaaagcg ttagtagcaa aagtgtctca gagggaagag atggttaaaa aatgtcttgg 4201 agaacttaca gaagtctgca aatctctggg gaaagttttt ggtgtccatt acttcaatat 4261 ttttaatacc gtcactctca agaagcttgc agaatcttta tcttctgatc ctgaggtttt 4321 gcttcaaatt gatggtgtta ctgaagacaa actggaaaaa tatggtgcgg aagtgatttc 4381 agtattacag aaatactctg aatggacatc gccagctgaa gacagttccc cagggataag 4441 cctgtccagc agcagaggcc ccggaagaag tgccgctgag gagcttgacg aggaaatacc 4501 cgtatcttcc cactactttg caagtaaaac cagaaatgaa aggaagagga aaaagatgcc 4561 agcctcccaa aggtctaaga ggagaaaaac tgcttccagt ggttccaagg caaagggggg 4621 gtctgccaca tgtagaaaga tatcttccaa aacgaaatcc tccagcatca ttggatccag 4681 ttcagcctca catacttctc aagcgacatc aggagccaat agcaaattgg ggattatggc 4741 tccaccgaag cctataaata gaccgtttct taagccttca tatgcattct cacaccacca 4801 ccaccaccac tgactcgagc atgcatctag agggccgcat catgtaatta gttatgtcac 4861 gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac 4921 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta 4981 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga 5041 aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag ctgcggccct 5101 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 5161 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 5221 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 5281 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 5341 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 5401 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 5461 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 5521 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 5581 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 5641 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 5701 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 5761 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 5821 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 5881 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 5941 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 6001 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 6061 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 6121 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 6181 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 6241 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 6301 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 6361 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 6421 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 6481 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 6541 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 6601 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 6661 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 6721 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 6781 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 6841 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 6901 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 6961 cctttttcaa tgggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt 7021 atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat 7081 gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca 7141 aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt 7201 ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc 7261 aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca 7321 aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg 7381 gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct 7441 tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta 7501 atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta 7561 ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc 7621 tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt 7681 agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga 7741 acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca 7801 gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat 7861 cttcgtttcc tgcaggtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat 7921 gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt 7981 tcttccttct gttcggagat taccgaatca aaaaaatttc aaggaaaccg aaatcaaaaa 8041 aaagaataaa aaaaaaatga tgaattgaaa agctagctta tcgatgataa gctgtcaaac 8101 atgagaatta attccacgga ctatagacta tacctagtat actccgtcta ctgtacgata 8161 cacttccgct caggtccttg tcctttaacg aggccttacc actcttttgt tactctattg 8221 atccagctca gcaaaggcag tgtgatctaa gattctatct tcgcgatgta gtaaaactag 8281 ctagaccgag aaagagacta gaaatgcaaa aggcacttct acaatggctg ccatcattat 8341 tatccgatgt gacgctgcag cttctcaatg atattcgaat acgctttgag gagatacagc 8401 ctaatatccg acaaactgtt ttacagattt acgatcgtac ttgttaccca tcattgaatt 8461 ttgaacatcc gaacctggga gttttccctg aaacagatag tatatttgaa cctgtataat 8521 aatatatagt ctagcgcttt acggaagaca atgtatgtat ttcggttcct ggagaaacta 8581 ttgcatctat tgcataggta atcttgcacg tcgcatcccc ggttcatttt ctgcgtttcc 8641 atcttgcact tcaatagcat atctttgtta acgaagcatc tgtgcttcat tttgtagaac 8701 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag 8761 aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt 8821 aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt 8881 tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt 8941 tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac 9001 tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt 9061 ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg 9121 actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa 9181 aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt 9241 gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct 9301 ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa 9361 tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt 9421 agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg 9481 atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat 9541 tcgcaatggg aagctccacc ccggttgata atcagaaaag ccccaaaaac aggaagattg 9601 tataagcaaa tatttaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt 9661 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa 9721 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa 9781 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac 9841 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga 9901 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa 9961 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc 10021 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tggggatcga 10081 tccactagtc ca //
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pJK1質粒載體應用舉例
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