质粒名称: | pUC57-Ter |
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出品公司: | Addgene |
目录编号: | 170634 |
存储实验室: | Sheng Yang |
载体骨架基本信息 |
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载体名称: | pUC57mini |
完整质粒大小: | 6420 bp |
载体类型: | 细菌表达 |
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: | Mini-Tn6677, Ter cassette |
物种来源: | Vibrio cholera |
克隆方法: | Gibson Cloning |
文献引用方法 |
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材料和方法部分: | pUC57-Ter was a gift from Sheng Yang (Addgene plasmid # 170634 ; http://n2t.net/addgene:170634 ; RRID:Addgene_170634) |
参考文献部分: | Programming Cells by Multicopy Chromosomal Integration Using CRISPR-Associated Transposases. Zhang Y, Yang J, Yang S, Zhang J, Chen J, Tao R, Jiang Y, Yang J, Yang S. CRISPR J. 2021 Jun;4(3):350-359. doi: 10.1089/crispr.2021.0018. 10.1089/crispr.2021.0018 PubMed 34152213 |
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载体序列
LOCUS pUC57-Ter 2881 bp ds-DNA circular SYN 04-AUG-2021 DEFINITION DNA donor for transposition. ACCESSION . VERSION . KEYWORDS pUC57-Ter SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2881) AUTHORS Zhang Y, Yang J, Yang S, Zhang J, Chen J, Tao R, Jiang Y, Yang J, Yang S TITLE Programming Cells by Multicopy Chromosomal Integration Using CRISPR-Associated Transposases. JOURNAL CRISPR J. 2021 Jun;4(3):350-359. doi: 10.1089/crispr.2021.0018. PUBMED 34152213 REFERENCE 2 (bases 1 to 2881) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 4, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..2881 /organism="synthetic DNA construct" /mol_type="other DNA" RBS 246..268 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" terminator 294..325 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" terminator 438..486 /label=fd terminator /note="central terminator from bacteriophage fd (Otsuka and Kunisawa, 1982)" terminator 548..596 /label=fd terminator /note="central terminator from bacteriophage fd (Otsuka and Kunisawa, 1982)" primer_bind complement(665..687) /label=pGEX 3' /note="pGEX vectors, reverse primer" rep_origin complement(1126..1714) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1206..1225) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1885..2745) /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 2508..2527 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2746..2850) /gene="bla" /label=AmpR promoter ORIGIN 1 ccaatgatgc gaaaaaaccc cgccgaagcg gggttttttg cgaaaaaaat agggataaca 61 gggtaatgtt taaactgttg atacaaccat aaaatgataa ttacacccat aaattgataa 121 ttatcacacc cataaattga tattgcctct tcatggtcta aacttcagta agtttacgac 181 attttcatga agaggtcatt tccgggaatt ccatggtcga catgcataag cttcctagga 241 ataattttgt ttaactttaa gaaggagata tacatatggc tagctaagca gaaagtcaaa 301 agcctccgac cggaggcttt tgactattgc tcaacaggta aggcgcgagg ttacgcgtct 361 cgagttaatt aaactagtga gctcggtacc cggggatcct ctagaggtcg aaattcacct 421 cgaaagcaag ctgataaacc gatacaatta aaggctcctt ttggagcctt tttttttgga 481 gattttcaac gtgaaaaaat tattattcgc aattccaagc taattcacct cgaaagcaag 541 ctgataaacc gatacaatta aaggctcctt ttggagcctt tttttttgga gattttcaac 601 gtgaaaaaat tattattcgc aattccaagc tctgcctcgc gcgtttcggt gatgacggtg 661 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg 721 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca 781 tgacccagtc acgtagcgat agcggagtgt atgggctcga tcccctcgga tccaattggt 841 ccttactgca gtagttttgc tgaaatactc gattcacaaa aatatcaact tatggttgtt 901 ttgtgagata tcaatatatg gttgttttgt ggttaagttg ctgattataa ataattatta 961 aatatcactt tatggttgca tcaacagtta acattaccct gttatcccta aaaaaaaaac 1021 cccgcccctg acagggcggg gttttttttt taatatcgga aagaacatgt gagcaaaagg 1081 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 1141 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 1201 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 1261 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 1321 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 1381 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 1441 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 1501 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 1561 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 1621 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 1681 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 1741 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 1801 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 1861 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 1921 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 1981 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 2041 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 2101 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 2161 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 2221 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 2281 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 2341 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 2401 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 2461 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 2521 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 2581 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 2641 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 2701 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 2761 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 2821 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 2881 c //
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