质粒名称: | pNP1 OR sfGFP |
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出品公司: | Addgene |
目录编号: | 107364 |
存储实验室: | James Collins |
载体骨架基本信息 |
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载体名称: | pNP1 |
载体出品公司: | New England Biolabs |
空载体大小: | 2202 bp |
完整质粒大小: | 3049 bp |
载体类型: | 细菌细胞表达 |
筛选标记: | 新霉素(G418) |
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: | OR gate + sfGFP |
插入大小: | 877 bp |
启动子: | T7 |
克隆方法: | Gibson Cloning |
5’测序引物 | CCCGCGAAATTAATACGACTCACTATAGG |
3’测序引物 | AGTGCCAAGCTTCCGGATATAGTTC |
文献引用方法 |
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材料和方法部分: |
pNP1 OR sfGFP was a gift from James Collins (Addgene plasmid # 107364 ; http://n2t.net/addgene:107364 ; RRID:Addgene_107364)
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参考文献部分: | BioBits Explorer: A modular synthetic biology education kit. Huang A, Nguyen PQ, Stark JC, Takahashi MK, Donghia N, Ferrante T, Dy AJ, Hsu KJ, Dubner RS, Pardee K, Jewett MC, Collins JJ. Sci Adv. 2018 Aug 1;4(8):eaat5105. doi: 10.1126/sciadv.aat5105. eCollection 2018 Aug. 10.1126/sciadv.aat5105 PubMed 30083608 |
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载体序列
LOCUS pNP1 OR sfGFP 3079 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses sfGFP (a GFP) in Ecoli off a T7 promoter when pNP1 OR input A and pNP1 OR input B are co-expressed. ACCESSION . VERSION . KEYWORDS pNP1 OR sfGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3079) AUTHORS Huang A, Nguyen PQ, Stark JC, Takahashi MK, Donghia N, Ferrante T, Dy AJ, Hsu KJ, Dubner RS, Pardee K, Jewett MC, Collins JJ TITLE BioBits Explorer: A modular synthetic biology education kit. JOURNAL Sci Adv. 2018 Aug 1;4(8):eaat5105. doi: 10.1126/sciadv.aat5105. eCollection 2018 Aug. PUBMED 30083608 REFERENCE 2 (bases 1 to 3079) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..3079 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 26..45 /label=T7 /note="T7 promoter, forward primer" promoter 26..44 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 847..894 /codon_start=1 /product="11th beta-strand of superfolder GFP" /label=GFP11 /note="associates with GFP(1-10), consisting of the first 10 beta-strands, to reconstitute fluorescent GFP (Cabantous et al., 2005; Kamiyama et al., 2016)" /translation="RDHMVLHEYVNAAGIT" CDS 895..918 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" primer_bind complement(1057..1075) /label=T7 Term /note="T7 terminator, reverse primer" terminator 1071..1118 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin complement(1336..1924) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1416..1435) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2096..2956) /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 2719..2738 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2957..3061) /gene="bla" /label=AmpR promoter ORIGIN 1 gctagtggtg ctagccccgc gaaattaata cgactcacta tagggatgaa cttaagatcg 61 cgctttatgg ttatgaaaca gaggagacat aacatgaaag cgcgaaacag aacaagaata 121 agtgcaaatg cttgttctat cgtcttgaaa cagaggagac aagacatgag aacaagcaac 181 ctggcggcag cgcaaaagat gagcaaaggt gaagaactgt ttaccggcgt tgtgccgatt 241 ctggtggaac tggatggcga tgtgaacggt cacaaattca gcgtgcgtgg tgaaggtgaa 301 ggcgatgcca cgattggcaa actgacgctg aaatttatct gcaccaccgg caaactgccg 361 gtgccgtggc cgacgctggt gaccaccctg acctatggcg ttcagtgttt tagtcgctat 421 ccggatcaca tgaaacgtca cgatttcttt aaatctgcaa tgccggaagg ctatgtgcag 481 gaacgtacga ttagctttaa agatgatggc aaatataaaa cgcgcgccgt tgtgaaattt 541 gaaggcgata ccctggtgaa ccgcattgaa ctgaaaggca cggattttaa agaagatggc 601 aatatcctgg gccataaact ggaatacaac tttaatagcc ataatgttta tattacggcg 661 gataaacaga aaaatggcat caaagcgaat tttaccgttc gccataacgt tgaagatggc 721 agtgtgcagc tggcagatca ttatcagcag aataccccga ttggtgatgg tccggtgctg 781 ctgccggata atcattatct gagcacgcag accgttctgt ctaaagatcc gaacgaaaaa 841 ggcacgcggg accacatggt tctgcacgaa tatgtgaatg cggcaggtat tacgtggagc 901 catccgcagt tcgaaaaata aaggatcccg ggaattctcg agtaaggtta acctgcagga 961 ggcctttaat taaggtggtg cggccgcgct agcggtcccg ggggatcgat ccggctgcta 1021 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac 1081 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg 1141 gaagcttggc actggccgac cggggtcgag cactgactcg ctgcgctcgg tcgttcggct 1201 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga 1261 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 1321 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 1381 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 1441 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 1501 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 1561 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 1621 cgccttatcc ggtaactatc gtcttgagtc caacccgcta agacacgact tatcgccact 1681 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 1741 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct 1801 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 1861 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 1921 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcaca 1981 gatccgggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 2041 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 2101 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 2161 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 2221 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 2281 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 2341 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 2401 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 2461 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 2521 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 2581 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 2641 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 2701 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 2761 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 2821 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 2881 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 2941 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 3001 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 3061 gcacatttcc ccgaaaagt //
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