质粒名称: | pAG426GPD-ccdB |
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出品公司: | 爱迪基因 |
目录编号: | 14156 |
存储实验室: | Susan Lindquist |
载体骨架基本信息 |
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载体名称: | pRS426 |
载体出品公司: | -- |
载体抗性: | 氯霉素和氨苄青霉素 |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 酵母表达;Gateway Destination |
筛选标记: | URA3 |
高拷贝/低拷贝: | -- |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | -- |
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: | Gateway克隆 |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | GPDPro_F |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pAG426GPD-ccdB was a gift from Susan Lindquist (Addgene plasmid # 14156 ; http://n2t.net/addgene:14156 ; RRID:Addgene_14156) |
买家导航
pAG426GPDccdB载体基本信息
pAG426GPDccdB载体图谱质粒图谱和多克隆位点信息
pAG426GPDccdB载体简介
pAG426GPDccdB载体序列
LOCUS pAG426GPD-ccdB 8306 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pAG426GPD-ccdB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8306) TITLE S cerevisiae Advanced Gateway Destination Vectors REFERENCE 2 (bases 1 to 8306) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8306 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(10..28) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" rep_origin 69..1411 /label=2u ori /note="yeast 2u plasmid origin of replication" protein_bind 401..448 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." promoter 1438..1542 /gene="bla" /label=AmpR promoter CDS 1543..2403 /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 complement(1761..1780) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2574..3162 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3063..3082 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3316..3333 /label=L4440 /note="L4440 vector, forward primer" protein_bind 3450..3471 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3486..3516 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3524..3540 /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)." primer_bind 3529..3551 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 3548..3564 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 3548..3564 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 3583..3603 /label=T3 /note="T3 promoter, forward primer" promoter 3585..3603 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 3628..4271 /gene="S. cerevisiae TDH3" /label=GAP promoter /note="promoter for glyceraldehyde-3-phosphate dehydrogenase" primer_bind 4165..4188 /label=GPDpro-F /note="S. cerevisiae GPD promoter, forward primer" primer_bind 4277..4293 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 4277..4293 /label=pBluescriptSK /note="For pBluescript vector" protein_bind 4301..4425 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 4450..4480 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" CDS 4534..5193 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(4600..4619) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 5535..5840 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 5719..5738 /label=ccdB-fwd /note="ccdB gene, forward primer" protein_bind complement(5881..6005) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(6038..6054) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6039..6055) /label=pBluescriptKS /note="For pBluescript vector" terminator 6057..6304 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(6073..6091) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind complement(6322..6341) /label=T7 /note="T7 promoter, forward primer" promoter complement(6323..6341) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6351..6368) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(6351..6367) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6360..6382) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 6508..6963 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6595..6614) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6805..6826 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(7094..7897) /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(7898..8113) /gene="S. cerevisiae URA3" /label=URA3 promoter primer_bind complement(8137..8156) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8256..8278 /label=pGEX 3' /note="pGEX vectors, reverse primer" ORIGIN 1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 61 cttagtatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg 121 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg 181 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac 241 gcatataagt acgcatttaa gcataaacac gcactatgcc gttcttctca tgtatatata 301 tatacaggca acacgcagat ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc 361 gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt 421 cctattctct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa 481 gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa aatattgcga 541 ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat 601 ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct 661 acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta 721 ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag 781 agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc 841 actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct 901 ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt 961 caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg 1021 acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa 1081 gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa 1141 aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt ctgttctgta 1201 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt 1261 acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg catttctgtt 1321 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt 1381 gttctacaaa atgaagcaca gatgcttcgt tcaggtggca cttttcgggg aaatgtgcgc 1441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 1501 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc 1561 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa 1621 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa 1681 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg 1741 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa 1801 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc 1861 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc 1921 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta 1981 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag 2041 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca 2101 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata 2161 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc 2221 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca 2281 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca 2341 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg 2401 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa 2461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 2521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 2581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 2641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 2701 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac 2761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 2821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 2881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 2941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 3001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 3061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 3121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 3181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc 3241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc 3301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa 3361 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac 3421 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc 3481 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa 3541 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa 3601 agggaacaaa agctggagct cagtttatca ttatcaatac tcgccatttc aaagaatacg 3661 taaataatta atagtagtga ttttcctaac tttatttagt caaaaaatta gccttttaat 3721 tctgctgtaa cccgtacatg cccaaaatag ggggcgggtt acacagaata tataacatcg 3781 taggtgtctg ggtgaacagt ttattcctgg catccactaa atataatgga gcccgctttt 3841 taagctggca tccagaaaaa aaaagaatcc cagcaccaaa atattgtttt cttcaccaac 3901 catcagttca taggtccatt ctcttagcgc aactacagag aacaggggca caaacaggca 3961 aaaaacgggc acaacctcaa tggagtgatg caacctgcct ggagtaaatg atgacacaag 4021 gcaattgacc cacgcatgta tctatctcat tttcttacac cttctattac cttctgctct 4081 ctctgatttg gaaaaagctg aaaaaaaagg ttgaaaccag ttccctgaaa ttattcccct 4141 acttgactaa taagtatata aagacggtag gtattgattg taattctgta aatctatttc 4201 ttaaacttct taaattctac ttttatagtt agtctttttt ttagttttaa aacaccagaa 4261 cttagtttcg acggattcta gaactagtgg atcccccatc acaagtttgt acaaaaaagc 4321 tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa 4381 acagactaca taatactgta aaacacaaca tatccagtca ctatggcggc cgcattaggc 4441 accccaggct ttacacttta tgcttccggc tcgtataatg tgtggatttt gagttaggat 4501 ccgtcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc 4561 accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct 4621 caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag 4681 aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct 4741 catccggaat tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac 4801 ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac 4861 cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa 4921 aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc 4981 tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc 5041 gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt 5101 caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa 5161 cagtactgcg atgagtggca gggcggggcg taaacgcgtg gatccggctt actaaaagcc 5221 agataacagt atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg 5281 tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca gtgacagttg 5341 acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag 5401 cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca 5461 ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa 5521 caggggctgg tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc 5581 tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc 5641 tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata 5701 tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta 5761 tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc 5821 tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtct gcaggtcgac 5881 catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc 5941 taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa 6001 gtggtgatgg gctgcaggaa ttcgatatca agcttatcga taccgtcgac ctcgagtcat 6061 gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa 6121 ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt 6181 attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc 6241 atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat 6301 ttgcggccgg tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc 6361 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca 6421 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa 6481 cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg 6541 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 6601 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 6661 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 6721 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 6781 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 6841 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 6901 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa 6961 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagggta 7021 ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt 7081 ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt 7141 ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa 7201 caataataat gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca 7261 atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt 7321 catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct 7381 tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca 7441 attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca 7501 aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg 7561 ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt 7621 ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg 7681 tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg 7741 gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac 7801 acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat 7861 gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg 7921 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac 7981 atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg 8041 agattaccga atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa 8101 atgatgaatt gaaaaggtgg tatggtgcac tctcagtaca atctgctctg atgccgcata 8161 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct 8221 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt 8281 ttcaccgtca tcaccgaaac gcgcga //
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pAG426GPDccdB载体质粒应用举例
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