质粒名称: | pSBY3_ligD-HygR |
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出品公司: | Addgene |
目录编号: | 104625 |
存储实验室: | Sheng Yang |
载体骨架基本信息 |
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载体名称: | pMV261 |
完整质粒大小: | 10039bp |
载体类型: | Bacterial Expression, CRISPR |
载体抗性: | Hygromycin |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | FnCpf1 |
基因突变: | FnCpf1 codon-optimized C.glutamicum |
启动子: | hsp60 |
克隆方法: | Gibson Cloning |
文献引用方法 |
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材料和方法部分: | pSBY3_ligD-HygR was a gift from Sheng Yang (Addgene plasmid # 104625 ; http://n2t.net/addgene:104625 ; RRID:Addgene_104625) |
参考文献部分: | A CRISPR-Cpf1-Assisted Non-Homologous End Joining Genome Editing System of Mycobacterium smegmatis. Sun B, Yang J, Yang S, Ye RD, Chen D, Jiang Y. Biotechnol J. 2018 Sep;13(9):e1700588. doi: 10.1002/biot.201700588. Epub 2018 Aug 6. 10.1002/biot.201700588 PubMed 30039929 |
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载体序列
LOCUS pSBY3_ligD-HygR 10039 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CRISPR system used to genome editing in Mycobacterium smegmatis, expresses Fn Cpf1 and crRNA targeting ligD and hygromysin resistance gene.. ACCESSION . VERSION . KEYWORDS pSBY3_ligD-HygR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10039) AUTHORS Sun B, Yang J, Yang S, Ye RD, Chen D, Jiang Y TITLE A CRISPR-Cpf1-Assisted Non-Homologous End Joining Genome Editing System of Mycobacterium smegmatis. JOURNAL Biotechnol J. 2018 Sep;13(9):e1700588. doi: 10.1002/biot.201700588. Epub 2018 Aug 6. PUBMED 30039929 REFERENCE 2 (bases 1 to 10039) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..10039 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 26..53 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 145..230 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind complement(438..457) /label=pENTR-R /note="pENTR vectors, reverse primer" primer_bind complement(522..541) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 3017..3605 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3506..3525 /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 9971..10017 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 gctagaaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttaat 61 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcgc 121 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca 181 acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt aaatcaccgt 241 tgcgtgtgac gtgctatcta caacagtaga aattcggatc cattatacct aggactgagc 301 tagctgtcaa tctagccacc tgacgtcggg ggggggggaa agccacgttg tgtctcaaaa 361 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc 421 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc 481 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga 541 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccc atgcgccaga 601 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag 661 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc 721 tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga 781 agaatatcct gattcaggtg aaaatagatg atcgaggatg ccagggcctt tcacgcccgc 841 cgctgctgag cgtccgccgc cgggcggccc ggggcgtcag gcgccggggg cggtgtccgg 901 cggcccccag aggaactgcg ccagttcctc cggatcggtg aagccggaga gatccagcgg 961 ggtctcctcg aacacctcga agtcgtgcag gaaggtgaag gcgagcagtt cgctggcgaa 1021 gtcctcggtc cgcttccact gcgccccgtc gagcagcgcg gccaggatct cgcggtcgcc 1081 ccggaaggcg ttgagatgca gttgcaccag gctgtagcgg gagtctcccg catagacgtc 1141 ggtgaagtcg acgatcccgg tgacctcggt cgcggccagg tccacgaaga tgttggtccc 1201 gtgcaggtcg ccgtggacga accggggttc gcggccggcc agcagcgtgt ccacgtccgg 1261 cagccagtcc tccaggcggt ccagcagccg gggcgagagg tagccccacc cgcggtggtc 1321 ctcgacggtc gccgcgcggc gttcccgcag cagttccggg aagacctcgg aatggggggt 1381 gagcacggtg ttcccggtca gcggcaccct gtgcagccgg ccgagcaccc ggccgagttc 1441 gcgggccagg gcgagcagcg cgttccggtc ggtcgtgccg tccatcgcgg accgccaggt 1501 ggtgccggtc atccggctca tcaccaggta gggccacggc caggctccgg tgccgggccg 1561 cagctcgccg cggccgagga ggcggggcac cggcaccggg gcgtccgcca ggaccgcgta 1621 cgcctccgac tccgacgcga ggctctccgg accgcaccag tgctcgccga acagcttgat 1681 caccgggccg ggctcgccga ccagtacggg gttggtgctc tcgccgggca cccgcagcac 1741 cggcggcacc ggcagcccga gctcctccag ggctcggcgg gccagcggct cccagaattc 1801 ctggtcgttc cgcaggctcg cgtaggaatc atccgaatca atacggtcga gaagtaacag 1861 ggattcttgt gtcacagcgg acctctattc acagggtacg ggccggctta attccgcacg 1921 gccggtcgcg acacggcctg tccgcaccgc ggatcaggcg ttgacgatga cgggctggtc 1981 ggccacgtcg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg 2041 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat 2101 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct 2161 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 2221 gatcaaagat tgttgatcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg 2281 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa 2341 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca 2401 agtctggaaa gaaatgcata atcttttgcc attctcaccg gattcagtcg tcactcatgg 2461 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt 2521 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg 2581 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga 2641 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa 2701 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcggct ttgttgaata 2761 aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt 2821 tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa agctctcacc 2881 aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcctg gtatgagtca 2941 gcaacacctt cttcacgagg cagacctcac tagttccatg agcgtcagac cccgtagaaa 3001 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 3061 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 3121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt 3181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 3241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 3301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 3361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 3421 ccacgcttcc cgaggggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 3481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 3541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 3601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 3661 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt 3721 gagctgatac cgctcgccgc agccgaacga ccgagcgcaa cgcgtgagcc caccagctcc 3781 gtaagttcgg gcgctgtgtg gctcgtaccc gcgcattcag gcggcagggg gtctaacggg 3841 tctaaggcgg cgtgtacggc cgccacagcg gctctcagcg gcccggaaac gtcctcgaaa 3901 cgacgcatgt gttcctcctg gttggtacag gtggttgggg gtgctcggct gtcgctggtg 3961 ttccaccacc agggctcgac gggagagcgg gggagtgtgc agttgtgggg tggcccctca 4021 gcgaaatatc tgacttggag ctcgtgtcgg accatacacc ggtgattaat cgtggtctac 4081 taccaagcgt gagccacgtc gccgacgaat ttgagcagct ctggctgccg tactggccgc 4141 tggcaagcga cgatctgctc gaggggatct accgccaaag ccgcgcgtcg gccctaggcc 4201 gccggtacat cgaggcgaac ccaacagcgc tggcaaacct gctggtcgtg gacgtagacc 4261 atccagacgc agcgctccga gcgctcagcg cccgggggtc ccatccgctg cccaacgcga 4321 tcgtgggcaa tcgcgccaac ggccacgcac acgcagtgtg ggcactcaac gcccctgttc 4381 cacgcaccga atacgcgcgg cgtaagccgc tcgcatacat ggcggcgtgc gccgaaggcc 4441 ttcggcgcgc cgtcgatggc gaccgcagtt actcaggcct catgaccaaa aaccccggcc 4501 acatcgcctg ggaaacggaa tggctccact cagatctcta cacactcagc cacatcgagg 4561 ccgagctcgg cgcgaacatg ccaccgccgc gctggcgtca gcagaccacg tacaaagcgg 4621 ctccgacgcc gctagggcgg aattgcgcac tgttcgattc cgtcaggttg tgggcctatc 4681 gtcccgccct catgcggatc tacctgccga cccggaacgt ggacggactc ggccgcgcga 4741 tctatgccga gtgccacgcg cgaaacgccg aatttccgtg caacgacgtg tgtcccggac 4801 cgctaccgga cagcgaggtc cgcgccatcg ccaacagcat ttggcgttgg atcacaacca 4861 agtcgcgcat ttgggcggac gggatcgtgg tctacgaggc cacactcagt gcgcgccagt 4921 cggccatctc gcggaagggc gcagcagcgc gcacggcggc gagcacagtt gcgcggcgcg 4981 caaagtccgc gtcagccatg gaggcattgc tatgagcgac ggctacagcg acggctacaa 5041 ccggcagccg actgtccgca aaaagcggcg cgtgaccgcc gccgaaggcg ctcgaatcac 5101 cggactatcc gaacgccacg tcgtccggct cgtggcgcag gaacgcagcg agtggctcgc 5161 cgagcaggct gcacgccgcg aacgcatccg cgcctatcac gacgacgagg gccactcttg 5221 gccgcaaacg gccaaacatt tcgggctgca tctggacacc gttaagcgac tcggctatcg 5281 ggcgaggaaa gagcgtgcgg cagaacagga agcggctcaa aaggcccaca acgaagccga 5341 caatccaccg ctgttctaac gcaattgggg agcgggtgtc gcgggggttc cgtggggggt 5401 tccgttgcaa cgggtcggac aggtaaaagt cctggtagac gctagttttc tggtttgggc 5461 catgcctgtc tcgttgcgtg tttcgttgcg cccgttttga ataccagcca gacgagacgg 5521 ggttctacga atcttggtcg ataccaagcc atttccgctg aatatcgggg agctcaccgc 5581 cagaatcggt ggttgtggtg atgtacgtgg cgaactccgt tgtagtgcct gtggtggcat 5641 ccgtggcgcg gccgcggtac cagatcttta aatctagacg gtgaccacaa cgcgcccgct 5701 ttgatcgggg acgtctgcgg ccgaccattt acgggtcttg ttgtcgttgg cggtcatggg 5761 ccgaacatac tcacccggat cggagggccg aggacaaggt cgaacgaggg gcatgacccg 5821 gtgcggggct tcttgcactc ggcataggcg agtgctaaga ataacgttgg cactcgcgac 5881 cggtgagtgc taggtcggga cggtgaggcc aggcccgtcg tcgcagcgag tggcagcgag 5941 gacaacttga gccgtccgtc gcgggcactg cgcccggcca gcgtaagtag cggggttgcc 6001 gtcacccggt gacccccgtt tcatccccga tccggaggaa tcacttcgca atgtccatct 6061 accaagagtt tgtgaataaa tactccctgt ccaagaccct ccgttttgag ctgatccccc 6121 aaggcaagac cctcgaaaac atcaaggcac gcggcctcat cctggatgac gaaaagcgcg 6181 ctaaggatta caagaaggca aagcagatca tcgacaagta ccaccagttc ttcatcgaag 6241 agatcctgtc ctccgtgtgc atctccgagg acctgctcca gaactactcc gatgtctact 6301 tcaagctcaa gaagtccgat gacgataacc tgcagaagga cttcaagtcc gctaaggata 6361 ccatcaagaa gcagatctcc gaatacatca aggattccga gaagttcaag aacctcttca 6421 accagaacct gatcgacgca aagaagggcc aggaatccga tctcatcctg tggctcaagc 6481 agtccaagga taacggcatc gagctcttca aggccaactc cgacatcacc gacatcgatg 6541 aagctctgga gatcatcaag tccttcaagg gctggaccac ctacttcaag ggcttccacg 6601 aaaaccgcaa gaacgtgtac tcctccaacg atatcccaac ctctatcatc taccgcatcg 6661 tcgacgataa cctgccaaag ttcctcgaaa acaaggcaaa gtacgagtcc ctgaaggata 6721 aggccccaga agctatcaac tacgagcaga tcaagaagga cctggccgaa gagctcacct 6781 tcgacatcga ttacaagacc tctgaagtga accagcgcgt cttctccctc gatgaagtgt 6841 tcgagatcgc caacttcaac aactacctga accagtccgg catcaccaag ttcaacacca 6901 tcatcggcgg caagttcgtc aacggcgaaa acaccaagcg caagggcatc aacgagtaca 6961 tcaacctcta ctcccagcag atcaacgata agaccctgaa gaagtacaag atgtccgtgc 7021 tcttcaagca gatcctgtcc gacaccgaat ccaagtcctt cgtcatcgac aagctggagg 7081 acgattccga tgtggtcacc accatgcagt ccttctacga acagatcgca gccttcaaga 7141 ccgtggaaga gaagtccatc aaggagaccc tctccctgct cttcgacgat ctgaaggctc 7201 agaagctgga tctctccaag atctacttca agaacgacaa gtccctgacc gatctctccc 7261 agcaggtctt cgacgattac tccgtgatcg gcaccgcagt cctggaatac atcacccagc 7321 agatcgcccc aaagaacctc gataacccat ccaagaagga acaggagctg atcgccaaga 7381 agaccgaaaa ggctaagtac ctgtccctcg agaccatcaa gctggctctc gaagagttca 7441 acaagcaccg cgacatcgat aagcagtgcc gcttcgaaga gatcctcgca aacttcgctg 7501 caatcccaat gatcttcgac gaaatcgcac agaacaagga taacctggcc cagatctcca 7561 tcaagtacca gaaccagggc aagaaggatc tgctccaggc ctccgctgag gacgatgtga 7621 aggcaatcaa ggacctgctc gatcagacca acaacctgct ccacaagctg aagatcttcc 7681 acatctccca gtccgaagac aaggccaaca tcctcgacaa ggatgagcac ttctacctgg 7741 tgttcgaaga gtgctacttc gaactcgcta acatcgtccc actgtacaac aagatccgca 7801 actacatcac ccagaagcca tactccgatg aaaagttcaa gctcaacttc gagaactcca 7861 ccctggcaaa cggctgggac aagaacaagg aaccagataa caccgccatc ctcttcatca 7921 aggacgataa gtactacctg ggcgtgatga acaagaagaa caacaagatc ttcgacgata 7981 aggccatcaa ggaaaacaag ggcgagggct acaagaagat cgtgtacaag ctgctcccag 8041 gcgctaacaa gatgctccca aaggtcttct tctccgcaaa gtccatcaag ttctacaacc 8101 catccgaaga tatcctgcgc atccgcaacc actccaccca caccaagaac ggctccccac 8161 agaagggcta cgaaaagttc gagttcaaca tcgaagactg ccgcaagttc atcgatttct 8221 acaagcagtc catctccaag cacccagagt ggaaggactt cggcttccgc ttctccgata 8281 cccagcgcta caactccatc gatgaattct accgcgaagt ggagaaccag ggctacaagc 8341 tgaccttcga aaacatctcc gagtcctaca tcgattccgt ggtcaaccag ggcaagctgt 8401 acctcttcca gatctacaac aaggacttct ccgcttactc caagggccgc ccaaacctgc 8461 acaccctcta ctggaaggca ctcttcgacg aacgcaacct gcaggatgtg gtctacaagc 8521 tcaacggcga agcagagctg ttctaccgca agcagtccat cccaaagaag atcacccacc 8581 cagccaagga agcaatcgcc aacaagaaca aggataaccc aaagaaggaa tccgtgttcg 8641 agtacgacct gatcaaggat aagcgcttca ccgaggacaa gttcttcttc cactgcccaa 8701 tcaccatcaa cttcaagtcc tccggcgcca acaagttcaa cgatgaaatc aacctgctcc 8761 tgaaggagaa ggctaacgac gtgcacatcc tgtccatcga tcgcggcgaa cgccacctcg 8821 cctactacac cctggtcgac ggcaagggca acatcatcaa gcaggacacc ttcaacatca 8881 tcggcaacga tcgcatgaag accaactacc acgacaagct ggccgctatc gagaaggacc 8941 gcgattccgc tcgcaaggat tggaagaaga tcaacaacat caaggaaatg aaggaaggct 9001 acctctccca ggtggtccac gaaatcgcta agctggtgat cgagtacaac gcaatcgtgg 9061 tcttcgaaga cctgaacttc ggcttcaagc gcggccgctt caaggtggag aagcaggtct 9121 accagaagct ggaaaagatg ctcatcgaga agctgaacta cctcgtgttc aaggacaacg 9181 aattcgataa gaccggcggc gtcctccgtg cataccagct gaccgcccca ttcgagacct 9241 tcaagaagat gggcaagcag accggcatca tctactacgt gccagctggc ttcacctcta 9301 agatctgccc agtgaccggc ttcgtcaacc agctctaccc aaagtacgaa tccgtctcca 9361 agtcccagga gttcttctcc aagttcgaca agatctgcta caacctggat aagggctact 9421 tcgaattctc cttcgactac aagaacttcg gcgataaggc agccaagggc aagtggacca 9481 tcgcatcctt cggctcccgc ctcatcaact tccgcaactc cgacaagaac cacaactggg 9541 atacccgcga agtgtaccca accaaggaac tggagaagct cctgaaggat tactccatcg 9601 aatacggcca cggcgagtgc atcaaggctg caatctgcgg cgaatccgac aagaagttct 9661 tcgcaaagct gacctctgtg ctcaacacca tcctgcagat gcgcaactcc aagaccggca 9721 ccgagctgga ttacctcatc tccccagtgg ccgacgtcaa cggcaacttc ttcgattccc 9781 gccaggctcc aaagaacatg ccacaggacg ctgatgcaaa cggcgcctac cacatcggtc 9841 tgaagggtct catgctcctg ggtcgcatca agaacaacca ggaaggcaag aagctgaatc 9901 tcgtcattaa gaacgaagaa tactttgaat ttgtccagaa ccgcaataac taagatcgac 9961 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatgcc 10021 atcatggccg cggtgatca //
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