质粒名称: | pLb-Cpf1-pGL3-U6-sgRNA |
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出品公司: | Addgene |
目录编号: | 107682 |
存储实验室: | Jia Chen |
载体骨架基本信息 |
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载体名称: | pGL3-Basic |
完整质粒大小: | 4897 bp |
载体类型: | 哺乳细胞表达 |
载体抗性: | 氨苄霉素 |
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: | 无 |
物种来源: | 光合细菌 Rhodopseudomonas palustris |
文献引用方法 |
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材料和方法部分: | pLb-Cpf1-pGL3-U6-sgRNA was a gift from Jia Chen (Addgene plasmid # 107682 ; http://n2t.net/addgene:107682 ; RRID:Addgene_107682) |
参考文献部分: | Base editing with a Cpf1-cytidine deaminase fusion. Li X, Wang Y, Liu Y, Yang B, Wang X, Wei J, Lu Z, Zhang Y, Wu J, Huang X, Yang L, Chen J. Nat Biotechnol. 2018 Mar 19. pii: nbt.4102. doi: 10.1038/nbt.4102. 10.1038/nbt.4102 PubMed 29553573 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pLb-Cpf1-pGL3-U6-sgRNA 4897 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses LbCpf1 crRNA in mammalian cells. ACCESSION . VERSION . KEYWORDS pLb-Cpf1-pGL3-U6-sgRNA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4897) AUTHORS Li X, Wang Y, Liu Y, Yang B, Wang X, Wei J, Lu Z, Zhang Y, Wu J, Huang X, Yang L, Chen J TITLE Base editing with a Cpf1-cytidine deaminase fusion. JOURNAL Nat Biotechnol. 2018 Mar 19. pii: nbt.4102. doi: 10.1038/nbt.4102. PUBMED 29553573 REFERENCE 2 (bases 1 to 4897) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..4897 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 61..301 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 61..81 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 232..251 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_feature 405..522 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 571..1081 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 1000..1018 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 1103..1702 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(1103..1122) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 1599..1619 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 1849..1970 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1860..1879) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1914..1933 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2218..2235) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2389..2977) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2469..2488) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3148..4008) /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 3771..3790 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4009..4113) /gene="bla" /label=AmpR promoter rep_origin 4140..4595 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4227..4246) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4437..4458 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 4726..4774 /note="synthetic polyadenylation signal" misc_feature 4788..4879 /label=pause site /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene" primer_bind 4828..4847 /label=RVprimer3 /note="pGL3 vector, forward primer" ORIGIN 1 gtgaacggat ctcgacggta tcgatcacga gactagcctc gagcggccgc ccccttcacc 61 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 121 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 181 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 241 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 301 cgaaacaccg aatttctact aagtgtagat cgagacccga gagagggtcg gtctcctttt 361 tttgaattct cgacctcgag acaaatggca gtattcatcc acaattttaa aagaaaaggg 421 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa 481 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac 541 agcagagatc cactttggcc gcggctcgag ggggttgggg ttgcgccttt tccaaggcag 601 ccctgggttt gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc 661 cgaccctggg actcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc 721 tacccttgtg ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt 781 gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc 841 agacggacag cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag 901 cggctgctca gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg 961 gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc 1021 tccggagcgc acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca 1081 gggggatcca ccggagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg 1141 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac 1201 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt 1261 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt 1321 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc 1381 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct 1441 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga 1501 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg 1561 cgccggggtg cccgccttcc tggaaacctc cgcgccccgc aacctcccct tctacgagcg 1621 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat 1681 gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag 1741 cgcacgaccc catgcatcgg tacctttaag accaatgact tacaaggcag ctgtagatct 1801 tagccacttt ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg 1861 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt 1921 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca 1981 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt 2041 aaaacctcta caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc 2101 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg 2161 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc 2221 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 2281 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 2341 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 2401 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 2461 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 2521 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 2581 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 2641 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 2701 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 2761 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 2821 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 2881 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 2941 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 3001 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 3061 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 3121 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 3181 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 3241 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggg acccacgctc 3301 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 3361 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 3421 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 3481 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 3541 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 3601 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 3661 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 3721 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 3781 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 3841 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 3901 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 3961 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 4021 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 4081 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 4141 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 4201 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 4261 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag 4321 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 4381 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 4441 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 4501 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 4561 cgcgaatttt aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa 4621 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg 4681 ataagtaagt aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta 4741 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc 4801 catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca 4861 ggtgccagaa catttctcta tcgataggta ccgatta //