质粒名称: | pSL1521 (pSPIN, pSC101* backbone) |
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出品公司: | Addgene |
目录编号: | 160729 |
存储实验室: | Samuel H. Sternberg |
载体骨架基本信息 |
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载体名称: | pSC101* |
载体出品公司: | Clontech |
完整质粒大小: | 12724bp |
载体类型: | 细菌表达 |
载体抗性: | Spectinomycin, 50 μg/mL |
生长温度: | 30℃ |
宿主菌: | DH5a |
拷贝数: | 低拷贝 |
插入基因信息 |
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插入基因名称: | VchCAST proteins, crRNA, and donor DNA |
物种来源: | V. cholerae |
启动子: | J23119 |
克隆方法: | Gibson Cloning |
5’测序引物 | CGCAGATCAGTTGGAAGAAT |
3’测序引物 | GCTAGTTATTGCTCAGCGG |
文献引用方法 |
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材料和方法部分: | pSL1521 (pSPIN, pSC101* backbone) was a gift from Samuel H. Sternberg (Addgene plasmid # 160729 ; http://n2t.net/addgene:160729 ; RRID:Addgene_160729) |
参考文献部分: | CRISPR RNA-guided integrases for high-efficiency, multiplexed bacterial genome engineering. Vo PLH, Ronda C, Klompe SE, Chen EE, Acree C, Wang HH, Sternberg SH. Nat Biotechnol. 2020 Nov 23. pii: 10.1038/s41587-020-00745-y. doi: 10.1038/s41587-020-00745-y. 10.1038/s41587-020-00745-y PubMed 33230293 |
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pSL1521载体质粒基本信息
pSL1521质粒图谱载体图谱和pSL1521载体序列质粒序列多克隆位点信息
pSL1521质粒载体简介
pSL1521质粒序列载体序列
LOCUS pSL1521 (pSPIN, pSC101* backbone) 12724 bp DNA circular SYN 25-AUG-2024 DEFINITION Single-plasmid V. cholerae CAST system. Encodes all proteins, crRNA, donor DNA; non-targeting crRNA has BsaI sites for cloning. Temperature-sensitive pSC101* backbone can be cured by 37 (o)C incubation.. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12724) AUTHORS Vo PLH, Ronda C, Klompe SE, Chen EE, Acree C, Wang HH, Sternberg SH TITLE CRISPR RNA-guided integrases for high-efficiency, multiplexed bacterial genome engineering. JOURNAL Nat Biotechnol. 2020 Nov 23. pii: 10.1038/s41587-020-00745-y. doi: 10.1038/s41587-020-00745-y. PUBMED 33230293 REFERENCE 2 (bases 1 to 12724) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.biofeng.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Biotechnol. 2020 Nov 23. pii: 10.1038/s41587-020-00745-y. doi: 10.1038/s41587-020-00745-y." FEATURES Location/Qualifiers source 1..12724 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 241..332 /gene="bla" /label=AmpR promoter CDS 333..1124 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" primer_bind 1286..1303 /label=L4440 /note="L4440 vector, forward primer" promoter 1310..1344 /label=J23119 promoter /note="bacterial promoter (Registry of Standard Biological Parts BBa_J23119)" CDS 8326..8337 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(9772..9790) /label=T7 Term /note="T7 terminator, reverse primer" terminator 9786..9833 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS 9988..10647 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(10054..10073) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" primer_bind complement(10847..10864) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(10847..10863) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(10856..10878) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 11470..11692 /label=pSC101 ori /note="low-copy replication origin that requires the Rep101 protein" CDS 11740..12690 /codon_start=1 /gene="rep101 encoding an A56V mutation" /product="temperature-sensitive version of the RepA protein needed for replication with the pSC101 origin (Armstrong et al., 1984)" /label=Rep101(Ts) /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI" ORIGIN 1 tcaagactaa caaacaaaag tagaacaact gttcaccgtt acatatcaaa gggaaaactg 61 tccataccca tggattcttc gtctgtttct actggtattg gcacaaacct gatatcgatg 121 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 181 attcaaatat gtatccgctc atgagacaat aaccctggta gcgaccgagt gagctagcta 241 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat 301 aaatgcttca ataatattga aaaaggaaga gtatgaggga agcggtgatc gccgaagtat 361 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg 421 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt 481 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc 541 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca 601 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg 661 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg 721 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg 781 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct 841 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt 901 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg 961 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg 1021 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg 1081 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataatgtcct gtcagcagac 1141 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 1201 taggtgaaga tccgcctgca tggataaccg tattaccgcc tttgagtgag ctgataccgc 1261 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagctt tgacagctag 1321 ctcagtccta ggtataatac tagcgtcgac gtggagatat accatgggtg aactgccgag 1381 taggtagctg ataacgagac ctctgtcttg tcagctaggg tggtctcgtg aactgccgag 1441 taggtagctg ataacggatc cgaattcgag cgaaggagat atacatatgt ttttgcaaag 1501 acctaaacct tacagcgatg aaagtttaga aagtttcttt atccgagtgg ctaacaaaaa 1561 tggctacggt gatgtccatc gcttcctaga agccactaaa cgattccttc aagacattga 1621 ccataatggc tatcaaacct ttccgactga tataactcgg ataaacccat actcagctaa 1681 aaacagttcc agcgcacgaa ctgcgtcatt cctgaaactt gcacaattga catttaatga 1741 accgccagag ctacttgggt tggcaattaa cagaacaaac atgaaatact cgccgtcaac 1801 tagcgcggtt gttagaggtg cagaagtctt tcctcgcagt ttactacgga cgcactccat 1861 cccctgctgt cctttgtgtc tgcgagaaaa tggctacgcc tcctaccttt ggcactttca 1921 ggggtacgaa tactgccaca gccataacgt acctttaatt accacttgta gctgtggtaa 1981 ggagtttgac taccgagtat ctgggttaaa gggcatttgc tgcaaatgca aggagcctat 2041 caccttaacc agcagggaga acggtcatga ggcagcgtgt actgtttcaa actggcttgc 2101 tggccatgaa tctaaacctc tgccaaatct tcctaaaagc taccgatggg gtttagttca 2161 ttggtggatg ggtattaaag atagcgagtt cgatcacttt tcgttcgttc aatttttctc 2221 aaactggcca aggtcattcc actcgataat cgaagatgaa gtagagttca accttgagca 2281 tgctgttgtc agcacgtctg aattacgact aaaagatctt cttggtcgat tgtttttcgg 2341 ttcaattcgg ttacctgagc ggaatcttca acacaatatc atccttggtg agcttctctg 2401 ctatttagaa aatcgtttat ggcaagacaa gggattaatc gccaacctca aaatgaacgc 2461 gttagaggcg actgtaatgt taaattgtag cctcgatcag attgcatcaa tggttgaaca 2521 acgcatcttg aagccaaatc gaaaaagcaa gcccaacagc cctcttgatg ttaccgatta 2581 tctatttcat ttcggcgata ttttctgtct ttggttagct gagttccaaa gcgatgagtt 2641 taaccgttcg ttttatgtgt cgaggtggta aatgcaaact ctgaaagaac taatcgcatc 2701 caatcctgac gacttaacaa ctgaacttaa gagagcattt cgtccactca caccgcatat 2761 tgcaattgat ggtaatgaac ttgacgcact gacgatatta gtcaatttaa ccgataagac 2821 tgatgatcag aaagacctgc tcgatcgagc caaatgcaag caaaaacttc gagatgaaaa 2881 atggtgggct agctgcataa attgcgttaa ttacagacaa agccataacc caaaattccc 2941 ggatatacgt tctgaaggcg tgattcgaac ccaagccctg ggtgaattac cgagtttcct 3001 actctcgtcc tcaaaaatcc caccatacca ttggtcatat agccatgatt caaaatacgt 3061 caacaaaagc gcattcctca ccaatgagtt ttgttgggat ggtgagatct catgtttggg 3121 tgagcttctt aaagatgcag atcacccact ttggaatact ttgaaaaagt taggttgttc 3181 tcaaaaaact tgcaaagcaa tggcaaaaca actagctgat attactctca cgactatcaa 3241 tgtcacactc gcaccaaatt acctgactca aatatctctc cccgatagtg atacatccta 3301 catttcactc tctcctgttg catcgctatc gatgcaaagc cactttcatc agaggctcca 3361 agatgaaaat cggcatagcg cgataacgcg gtttagccga actaccaaca tgggggtcac 3421 tgcgatgaca tgtggcggcg catttagaat gttaaagtct ggcgctaagt tttccagccc 3481 ccctcatcac cgattaaaca gtaaacgaag ttggttgacg tcagagcatg ttcagtcatt 3541 aaaacagtac cagcgcctca acaaaagcct catacctgaa aactctcgga ttgcactccg 3601 tagaaaatac aaaatcgagc ttcaaaatat ggtcagatct tggtttgcaa tgcaagacca 3661 tacccttgat tcgaacatac ttatccaaca tttgaatcat gacctatctt acttaggagc 3721 cacaaaacgt tttgcatacg atccagcgat gaccaagctc tttactgagc ttttgaaacg 3781 agagttatca aattcaatca ataatggtga gcaacacact aatggatcgt ttttagtcct 3841 accgaatatc agggtttgtg gcgcaacagc tttaagctcc ccggtaacgg tggggattcc 3901 atcacttaca gctttctttg gcttcgttca cgcatttgaa cggaatataa atcgcaccac 3961 ctcatcgttt cgtgttgaat cctttgcgat atgcgtccat caactacatg tcgagaagcg 4021 aggtttgaca gcagagtttg tggaaaaagg cgacgggact atatccgctc ccgcgacccg 4081 ggatgactgg cagtgtgatg tcgtatttag ccttattttg aacaccaact ttgctcaaca 4141 tattgaccaa gatacgttag ttacatcact accaaagcga ttggctcggg gttcagcaaa 4201 aattgcgatt gatgacttta aacatatcaa ctcattctcg acattagaaa cagcgatcga 4261 atctctgcca atagaagctg gtaggtggtt atcactttac gcacagtcaa acaataatct 4321 aagtgatcta ttagcagcca tgacagagga ccatcagctc atggcaagct gcgtcggtta 4381 ccacttgtta gaagagccca aagataaacc aaactccctc agaggttaca aacacgctat 4441 cgccgagtgc atcattggac tcattaactc aatcaccttt agctcagaga ctgatcccaa 4501 cacaatcttt tggtcgctaa agaactatca aaactaccta gtggtacagc caaggagtat 4561 caacgatgaa actaccgaca aatctagcct atgagcgctc tatcgaccca tcagatgtct 4621 gtttttttgt cgtctggccc gatgatagaa aaacaccttt aacctacaat tctcgtactc 4681 tgctcgggca aatggaagcg gcatcattag cctatgatgt ctcaggtcaa ccaataaaaa 4741 gtgccaccgc tgaggcgtta gctcaaggga accctcatca agttgatttc tgccacgttc 4801 catacggtgc gagtcatatt gaatgcagtt tctccgtctc gttttcttct gaactacgtc 4861 aaccatataa gtgtaactca agcaaagtta aacaaacgct agtgcaatta gtcgagctct 4921 acgaaacgaa aatcggctgg actgagctag caacccgata tttgatgaac atttgcaacg 4981 gtaaatggct gtggaaaaat acccgtaaag cctattgctg gaacattgta cttacacctt 5041 ggccttggaa cggggaaaag gttggatttg aagatattcg tactaactac acctcacggc 5101 aagactttaa aaataataaa aattggtctg ctatagttga aatgatcaaa accgcatttt 5161 ctagtactga tgggctggcg atatttgaag tcagggccac cttgcacttg ccaacgaatg 5221 ctatggtgcg gccaagccaa gttttcacag aaaaagaaag tggcagtaaa agtaaatcta 5281 aaactcaaaa cagtcgagtt tttcagagta caactattga tggtgaacga tcgccaatac 5341 taggggcctt taaaacggga gcagctattg caaccattga cgactggtat cctgaagcca 5401 ctgagccact aagggtcgga cggtttgggg ttcatcgcga agatgtcact tgctaccgtc 5461 atccgtctac cggaaaagat tttttctcga tattacaaca agcagagcac tatattgaag 5521 tgttgagcgc caacaaaact cccgctcaag aaactatcaa cgacatgcac tttttaatgg 5581 ctaacctgat taagggtggg atgttccagc ataaaggaga ctgactgtga aatggtatta 5641 taagacaatc acctttctgc cagagttgtg caacaacgag tcactggctg caaagtgtct 5701 ccgcgttctg catggattta actatcagta tgagacacga aatatcggcg tttcatttcc 5761 gctttggtgt gatgcaacgg ttggaaaaaa gatttcattt gtcagcaaga acaagataga 5821 actcgactta ctacttaaac aacactattt cgtccaaatg gaacaacttc aatactttca 5881 tatatccaac actgttctcg tcccagaaga ttgtacatac gtttccttta gacgctgtca 5941 atctatagat aagctcacag cagcagggct ggcaaggaaa atcagacgcc tggagaaacg 6001 tgctctatct agaggcgagc aatttgaccc atcatctttt gctcaaaaag agcatactgc 6061 aatagcgcac taccactcac ttggggagtc cagcaaacag acgaaccgca actttcgact 6121 caatatcagg atgctctcgg agcaaccccg tgaggggaac tcgattttta gtagctatgg 6181 cttatcaaat tcagaaaact cgtttcagcc tgtaccctta atctgacctt taataaggag 6241 atataccatg gcgacaagtt tacctacgcc ctcagcaatt acgacttcgg cgttagagta 6301 tgcattccat actcccgctc gcaatctaac gaaatctcgc ggaaaaaaca ttcatcgtta 6361 tgtcagtgta aagatgagta agaggattac ggtagaatct actctagagt gtgatgcctg 6421 ctatcacttt gattttgagc caagtattgt tcgcttttgc gctcaaccga ttcgattttt 6481 atattatctc aatggtcagt ctcactccta tgttcctgac tttctagttc aatttgatac 6541 caacgagttt gttctatatg aagtaaagtc agcttatgct aagaacaaac ctgattttga 6601 tgttgaatgg gaggcgaaag taaaagcagc aactgaacta gggctagaat tggagcttgt 6661 tgaagagagt gatattaggg atacggttgt attaaataat cttaagcgca tgcatcgtta 6721 tgcttcgaaa gatgagctga ataacgtaca taactctctc ttaaaaataa taaagtacaa 6781 tggcgcccaa tctgcaagat gcttgggaga acagttgggt ttaaaaggcc gaactgtttt 6841 accaattttg tgcgatttgc tgtcaaggtg tttactcgat acacgtttgg ataagcctct 6901 atctcttgaa tctcgatttg agttggccag ttatggctaa gaaagggttc tcaagtttcc 6961 atagaaaagc agtctcttcc caagatacgc ttgaatcgat agagcttgtc tctagcgcta 7021 attgtctaga aagtgttacg tatcaagata tatcagcatt tcccgaaaca attgcggtag 7081 agattaattt ccgattaagc attcttcgct ttttagcgcg aaagtgcgaa accattgtgg 7141 ccaaatcaat tgaaccacat cgtgtagagc tacagcaaaa ctatagtaga aaaataccca 7201 gtgcaataac gatatatcga tggtggcttg cttttcgaaa atcagactac aaccccatta 7261 gcttagcacc taatatcaag gatagaggta atagagagac aaaagtgtca acagttgttg 7321 attctattat ggaacaggca gttgaaagag ttatatctgg acgaaaagtc aatgttagct 7381 ctgcatataa acgtgttcga cgaaaagttc gtcaatacaa tctaactcat ggaacgaaat 7441 acacgtatcc taagtacgaa tctgtaagaa agcgagtaaa aaagaaaacc ccatttgagt 7501 tattagccgc agggaaaggg gagagagtag ctaagagaga gtttcgccga atgggaaaaa 7561 agatcctcac gtctagcgtg ctagagaggg ttgaaataga tcacactgtc gttgaccttt 7621 ttgcagtaca cgaagagtat cgaatcccat tgggccgacc ttggcttact caattggttg 7681 attgttacag taaagctgta atcggttttt atttaggttt cgagcctcct agctatgtgt 7741 cggtttccct tgcacttaag aatgcaatac aacgcaaaga tgacttaatc tcctcgtatg 7801 aatcgatcga gaatgaatgg ctatgttatg gcatcccaga cctactcgta actgataatg 7861 gtaaagagtt tttgtcgaaa gcatttgatc aagcatgtga atcactattg atcaatgtgc 7921 atcaaaataa agttgagacg cccgacaaca aacctcatgt tgaacgtaac tacgggacta 7981 ttaatacttc tctgttagac gatttacctg ggaaatcctt cagccagtac cttcaaagag 8041 aagggtacga ctctgtggga gaagctaccc ttacactcaa tgagattaga gaaatttact 8101 taatttggtt ggtggatatt tatcataaaa aacccaatca gagaggcact aattgtccta 8161 atgttgcgtg gaaaaagggt tgtcaagaat gggaaccaga ggagttctct ggttctaaag 8221 acgaattaga ctttaaattt gctattgttg attacaaaca acttactaaa gtagggataa 8281 ctgtctacaa agaactgagt tatagcaatg accgtttagc tgaatataga gggaagaaag 8341 gaaaccataa agttcagttc aagtataacc ctgagtgtat ggcagttatt tgggtgttgg 8401 atgaggatat gaatgagtac tttacagtta atgcgattga ctacgaatat gcaagtagag 8461 tatcactttg gcaacataaa tataacatga aatatcaagc agaactaaat tcagcagaat 8521 atgatgagga caaggaaatt gatgcagaaa taaaaattga agaaatcgca gatcgttcaa 8581 ttgttaagac taacaaaatc agagctcgga ggcgtggcgc taggcatcaa gagaatagcg 8641 caagggctaa gtcaatcagt aatgcgaacc cggcctcgat acaaaaacat gaagatgaaa 8701 tcgttagtgc agataatgac gattgggata ttgattatgt ctgagaaatc gtcaaatgag 8761 tgaaacgcgt gaggctcgta tatcaagagc taaaagggca tttgtatcca caccgagcgt 8821 taggaaaatc ttgagttaca tggatagatg tagagatcta tcagacctag agtctgagcc 8881 tacatgcatg atggtctatg gtgcttcggg tgtaggtaaa acgaccgtca tcaagaaata 8941 cttaaatcaa aacagaagag agtccgaagc cgggggcgat ataataccgg ttttgcatat 9001 tgagttgcca gacaatgcga agccagtaga tgcagcaagg gaattgctgg ttgaaatggg 9061 tgacccgcta gcactttatg aaactgactt agctagattg acgaaaagac tgactgaatt 9121 aatccctgcg gtcggcgtga agctgattat tatcgatgag ttccaacatt tggtggaaga 9181 aaggtcaaat cgggttctta cccaagtagg taattggcta aaaatgatac ttaacaaaac 9241 gaaatgtcca attgttatat ttggtatgcc atactcaaaa gttgtactgc aagcaaactc 9301 gcaacttcac gggcgatttt ccattcaggt tgaactgcgc ccctttagct accagggagg 9361 tagaggtgta tttaaaactt ttttggaata ccttgataaa gccctacctt ttgaaaaaca 9421 ggctggctta gccaacgaaa gtttgcagaa aaaattgtat gcattctctc agggaaacat 9481 gcgttcgttg agaaacctta tttatcaagc atctatcgaa gcaattgata atcagcatga 9541 gacgataacc gaagaagatt tcgtttttgc atcgaagttg acatcgggcg ataaacccaa 9601 ctcatggaaa aatccttttg aggagggtgt tgaggtaaca gaagatatgt tacgaccgcc 9661 accaaaagat attggttggg aggactattt gagacattca accccgagag tgagtaaacc 9721 aggtagaaat aaaaactttt tcgaataacc taggctgctg cgctgctgcc accgctgagc 9781 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac 9841 ctcaggcatt tgttgttgat acaaccataa aatgataatt acacccataa attgataatt 9901 atcacaccca taaattgata ttgcctcttc atggtctaaa cttcagtaag tttacgacat 9961 tttcctcgag gtcatttccg gggatccatg gagaaaaaaa tcactggata taccaccgtt 10021 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt 10081 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat 10141 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg 10201 gagttccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt 10261 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac 10321 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg 10381 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg 10441 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc 10501 actatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt 10561 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac 10621 tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttctaga 10681 gtccttactg cagtagtttt gctgaaatac tcgattcaca aaaatatcaa cttatggttg 10741 ttttgtgaga tatcaatata tggttgtttt gtggttaagt tgctgattat aaataattat 10801 taaatatcac tttatggttg catcaacagg taccgagctc gaattcactg gccgtcgttt 10861 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gccagttcac 10921 cgacaaacga aagctgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac 10981 tgagcgtcag accccgttga tgataccgct gccttactgg gtgcattagc cagtctgaat 11041 gacctgtcac gggataatcc gaagtggtca gactggaaaa tcagagggca ggaactgctg 11101 aacagcaaaa agtcagatag caccacatag cagacccgcc ataaaacgcc ctgagaagcc 11161 cgtgacgggc ttttcttgta ttatgggtag tttccttgca tgaatccata aaaggcgcct 11221 gtagtgccat ttacccccat tcactgccag agccgtgagc gcagcgaact gaatgtcacg 11281 aaaaagacag cgactcaggt gcctgatggt cggagacaaa aggaatattc agcgatttgc 11341 ccgattgcgg ccgcaaccga gcttgcgagg gtgctactta agcctttagg gttttaaggt 11401 ctgttttgta gaggagcaaa cagcgtttgc gacatccttt tgtaatactg cggaactgac 11461 taaagtagtg agttatacac agggctggga tctattcttt ttatcttttt ttattctttc 11521 tttattctat aaattataac cacttgaata taaacaaaaa aaacacacaa aggtctagcg 11581 gaatttacag agggtctagc agaatttaca agttttccag caaaggtcta gcagaattta 11641 cagataccca caactcaaag gaaaaggact agtaattatc attgactagc ccatctcaat 11701 tggtatagtg attaaaatca cctagaccaa ttgagatgta tgtctgaatt agttgttttc 11761 aaagcaaatg aactagcgat tagtcgctat gacttaacgg agcatgaaac caagctaatt 11821 ttatgctgtg tggcactact caaccccacg attgaaaacc ctacaaggaa agaacggacg 11881 gtatcgttca cttataacca atacgttcag atgatgaaca tcagtaggga aaatgcttat 11941 ggtgtattag ctaaagcaac cagagagctg atgacgagaa ctgtggaaat caggaatcct 12001 ttggttaaag gctttgagat tttccagtgg acaaactatg ccaagttctc aagcgaaaaa 12061 ttagaattag tttttagtga agagatattg ccttatcttt tccagttaaa aaaattcata 12121 aaatataatc tggaacatgt taagtctttt gaaaacaaat actctatgag gatttatgag 12181 tggttattaa aagaactaac acaaaagaaa actcacaagg caaatataga gattagcctt 12241 gatgaattta agttcatgtt aatgcttgaa aataactacc atgagtttaa aaggcttaac 12301 caatgggttt tgaaaccaat aagtaaagat ttaaacactt acagcaatat gaaattggtg 12361 gttgataagc gaggccgccc gactgatacg ttgattttcc aagttgaact agatagacaa 12421 atggatctcg taaccgaact tgagaacaac cagataaaaa tgaatggtga caaaatacca 12481 acaaccatta catcagattc ctacctacat aacggactaa gaaaaacact acacgatgct 12541 ttaactgcaa aaattcagct caccagtttt gaggcaaaat ttttgagtga catgcaaagt 12601 aagtatgatc tcaatggttc gttctcatgg ctcacgcaaa aacaacgaac cacactagag 12661 aacatactgg ctaaatacgg aaggatctga ggttcttatg gctcttgtat ctatcagtga 12721 agca //
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pSL1521质粒载体应用举例
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