质粒名称: | pCgsgRNA_crtYf |
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出品公司: | Addgene |
目录编号: | 153338 |
存储实验室: | Sheng Yang |
载体骨架基本信息 |
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载体名称: | pJYS2_crtYf |
完整质粒大小: | 4214bp |
载体类型: | Bacterial Expression, CRISPR |
载体抗性: | Spectinomycin |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 未知 |
插入基因信息 |
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插入基因名称: | crRNA targeting crtYf |
克隆方法: | Gibson Cloning |
5’测序引物 | 未知 |
文献引用方法 |
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材料和方法部分: | pCgsgRNA_crtYf was a gift from Sheng Yang (Addgene plasmid # 153338 ; http://n2t.net/addgene:153338 ; RRID:Addgene_153338) |
参考文献部分: | De Novo Engineering of Corynebacterium glutamicum for l-Proline Production. Zhang J, Qian F, Dong F, Wang Q, Yang J, Jiang Y, Yang S. ACS Synth Biol. 2020 Jul 17;9(7):1897-1906. doi: 10.1021/acssynbio.0c00249. Epub 2020 Jul 6. 10.1021/acssynbio.0c00249 PubMed 32627539 |
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LOCUS pCgsgRNA_crtYf 4214 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Double-plasmid-based CRISPR-Cas9 system in Corynebacterium glutamicum; rep oriVC. glutamicum; pMB1 oriVE. Coli; Pj23119-crRNA targeting crtYf; Spcr. ACCESSION . VERSION . KEYWORDS pCgsgRNA_crtYf SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4214) AUTHORS Zhang J, Qian F, Dong F, Wang Q, Yang J, Jiang Y, Yang S TITLE De Novo Engineering of Corynebacterium glutamicum for l-Proline Production. JOURNAL ACS Synth Biol. 2020 Jul 17;9(7):1897-1906. doi: 10.1021/acssynbio.0c00249. Epub 2020 Jul 6. PUBMED 32627539 REFERENCE 2 (bases 1 to 4214) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..4214 /organism="synthetic DNA construct" /mol_type="other DNA" misc_RNA 2259..2334 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" primer_bind complement(2416..2433) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2587..3175) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2667..2686) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ataaactatt taaataacag attaaaaaaa ttataaaaaa attgaaaaaa tggtggaaac 61 acttttttca atttttttgt tttattattt aatatttggg aaatattcat tctaattggt 121 aatcagattt tagaaaacaa taaacccttg catatgatat cgatgtacag atcttttttt 181 aaggcagtta ttggtgccct tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg 241 agatttcgat tccaccgccg ccttctatga aagccatcaa tcctgcctat ttgccacgtt 301 taacaaggta gttaagcgtt catttacgaa gaaaacacga taagctgcac aaatacctga 361 aaaagttgaa cgccccgtga gcgggaactc acagggcgtc ggctaacccc cagtcatcag 421 ctgggagaaa gcactcaaga catgactcta gccgatccgc aggacacagt cacagctagc 481 gcgtggaaat tttccgccga tctgttcgac acccaccccg aactagcgct gcgctcacgc 541 ggctggacgg cagaagatcg ccgcgaactg ctcgctcacc tgggacgcga aagcttccag 601 ggcagcaaga caagagattt cgcgagcgcc tggattaaaa acccggatac cggcgaaacc 661 caaccaaagc tctaccgggc tggctcaaaa gcgctgacgc ggtgccagta cgttgcgctg 721 acgcacgcgc aacatgccgc ggtgatcgtg cttgacatcg atgtgcccag ccaccaggcc 781 ggcgggaaga ttgagcacgt aaacccgcag gtctacgcga ttttagagaa atgggcacgc 841 ctagaaaaag cgccggcttg gatcggcgtg aatccgctga gcgggaaatg ccagctcatc 901 tggctcattg acccggtgta tgccgcagca ggtaaaacca gcccaaatat gcgcctgctg 961 gctgcaacga cggaagaaat gactcgtgtt ttcggcgctg accaggcttt ttcgcatagg 1021 ctgagccggt ggccgctgca cgtctcagac gatccgacag cctataaatg gcactgccag 1081 catgatcgtg tggatcggct ggccgaccta atggagattg ctcgaacgat gaccggatca 1141 cagaagccga aaaagtacat tgagcaggac ttttccagcg gacgcgcccg cattgaagcg 1201 gcacaacgcg ccaccgcaga agccaaggcg ctagcgattt tggacgcgag cctgccgagc 1261 gccctggacg cgtccggcga cctgatcgac ggcgtgcgag tgctctggac aaatccagag 1321 cgcgcagcgc gcgacgagac cgcgtttcgc cacgcgttga ccgtgggata ccagctcaaa 1381 gctgctggtg agcgcctaaa agatgccaag atcatcgacg cgtatgaagt ggcgtacaac 1441 gttgcccagg cggtcggtgc agacggccgg gagccggatc ttcccgccat gcgtgatcgc 1501 ctgacgatgg cgcgtcgtgt gcgcggctac gtggctaaag gccagccagt cgtccctgct 1561 cgtcgggtgg aaacgcagag cagccgaggg cggaaagctc tagcgacgat ggggcgacgg 1621 ggcgcagcta catcgaatgc acgcagatgg gctgacccag aaagtaagta tgcgcaggag 1681 acgcgacagc gattagcgga agcaaacaaa cgccgagaaa tgacaggcga gttgctcgaa 1741 cttcgcgtca aaactgcgat cctggatgcc cgttctcaat cggttgctga tccctcgact 1801 cgtgagcttg caggcgaact aggtgtcagt gaaaggcgca tccaacaagt cagaaaggca 1861 cttggaatgg aagctaaacg cggccgtcca cgggctgaaa actaataaac gaaacaccgt 1921 cagcagaaaa cggttccccc ctttaggggt cccgtccttg ctctggctct cacttgccct 1981 caccctccgc tatccacggg ctgaaaacta ataaacgaaa caccgtcagc agaaaacggt 2041 tccccccctt tagggtgtct cgctcctagc tctgatccct ccccggttcc tccccggcct 2101 gatttttaag ggggctcacg ctgtcggcag agaacggttc ccccgccttc tgctctggct 2161 cttcctcgac tccctccccc tcaaaaatct cctcgagtct agattgacag ctagctcagt 2221 cctaggtata atggatccct gcgtttaaac atatttccgt tttagagcta gaaatagcaa 2281 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt 2341 taagcttcat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag 2401 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 2461 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 2521 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 2581 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 2641 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 2701 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 2761 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 2821 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 2881 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 2941 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 3001 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 3061 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 3121 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 3181 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 3241 tttggtcatg aactaataac gtaacgtgac tggcaagaga tatttttaaa acaatgaata 3301 ggtttacact tactttagtt ttatggaaat gaaagatcat atcatatata atctagaata 3361 aaattaacta aaataattat tatctagata aaaaatttag aagccaatga aatctataaa 3421 taaactaaat taagtttatt taattaacaa ctatggatat aaaataggta ctaatcaaaa 3481 tagtgaggag gatatatttg aatacatacg aacaaattaa taaagtgaaa aaaatacttc 3541 ggaaacattt aaaaaataac cttattggta cttacatgtt tggatcagga gttgagagtg 3601 gactaaaacc aaatagtgat cttgactttt tagtcgtcgt atctgaacca ttgacagatc 3661 aaagtaaaga aatacttata caaaaaatta gacctatttc aaaaaaaata ggagataaaa 3721 gcaacttacg atatattgaa ttaacaatta ttattcagca agaaatggta ccgtggaatc 3781 atcctcccaa acaagaattt atttatggag aatggttaca agagctttat gaacaaggat 3841 acattcctca gaaggaatta aattcagatt taaccataat gctttaccaa gcaaaacgaa 3901 aaaataaaag aatatacgga aattatgact tagaggaatt actacctgat attccatttt 3961 ctgatgtgag aagagccatt atggattcgt cagaggaatt aatagataat tatcaggatg 4021 atgaaaccaa ctctatatta actttatgcc gtatgatttt aactatggac acgggtaaaa 4081 tcataccaaa agatattgcg ggaaatgcag tggctgaatc ttctccatta gaacataggg 4141 agagaatttt gttagcagtt cgtagttatc ttggagagaa tattgaatgg actaatgaaa 4201 atgtaaattt aact //
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