质粒名称: | pRE57-Ter |
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出品公司: | Addgene |
目录编号: | 170635 |
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: | 2811 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: | pRE57-Ter was a gift from Sheng Yang (Addgene plasmid # 170635 ; http://n2t.net/addgene:170635 ; RRID:Addgene_170635) |
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 pRE57-Ter 2811 bp ds-DNA circular SYN 04-AUG-2021 DEFINITION DNA donor for transposition. ACCESSION . VERSION . KEYWORDS pRE57-Ter SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2811) 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 2811) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 4, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..2811 /organism="synthetic DNA construct" /mol_type="other DNA" RBS 176..198 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" terminator 224..255 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" terminator 368..416 /label=fd terminator /note="central terminator from bacteriophage fd (Otsuka and Kunisawa, 1982)" terminator 478..526 /label=fd terminator /note="central terminator from bacteriophage fd (Otsuka and Kunisawa, 1982)" primer_bind complement(595..617) /label=pGEX 3' /note="pGEX vectors, reverse primer" rep_origin complement(1056..1644) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1136..1155) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1815..2675) /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 2438..2457 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2676..2780) /gene="bla" /label=AmpR promoter ORIGIN 1 ccaatgatgc gaaaaaaccc cgccgaagcg gggttttttg cgaaaaaaat agggataaca 61 gggtaatgtt taaactgttg atacaaccat aaaatgataa ttacacccat aaattgataa 121 ttatcacacc cacgggaatt ccatggtcga catgcataag cttcctagga ataattttgt 181 ttaactttaa gaaggagata tacatatggc tagctaagca gaaagtcaaa agcctccgac 241 cggaggcttt tgactattgc tcaacaggta aggcgcgagg ttacgcgtct cgagttaatt 301 aaactagtga gctcggtacc cggggatcct ctagaggtcg aaattcacct cgaaagcaag 361 ctgataaacc gatacaatta aaggctcctt ttggagcctt tttttttgga gattttcaac 421 gtgaaaaaat tattattcgc aattccaagc taattcacct cgaaagcaag ctgataaacc 481 gatacaatta aaggctcctt ttggagcctt tttttttgga gattttcaac gtgaaaaaat 541 tattattcgc aattccaagc tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg 601 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 661 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc 721 acgtagcgat agcggagtgt atgggctcga tcccctcgga tccaattggt ccttactgca 781 gtagttttgc tgaaatactc gattcacaaa aatatcaact tatggttgtt ttgtgagata 841 tcaatatatg gttgttttgt ggttaagttg ctgattataa ataattatta aatatcactt 901 tatggttgca tcaacagtta acattaccct gttatcccta aaaaaaaaac cccgcccctg 961 acagggcggg gttttttttt taatatcgga aagaacatgt gagcaaaagg ccagcaaaag 1021 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 1081 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 1141 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 1201 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 1261 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 1321 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 1381 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 1441 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 1501 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 1561 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 1621 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 1681 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 1741 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 1801 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 1861 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 1921 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 1981 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 2041 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 2101 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 2161 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 2221 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 2281 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 2341 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 2401 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 2461 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 2521 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 2581 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 2641 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 2701 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 2761 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c //
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