质粒名称: | pDonorGFP(Ptr) |
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出品公司: | Addgene |
目录编号: | 175578 |
存储实验室: | Sheng Yang |
载体骨架基本信息 |
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载体名称: | pACYC |
完整质粒大小: | 4606 bp |
载体类型: | 细菌表达 |
载体抗性: | Chloramphenicol |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 低拷贝 |
插入基因信息 |
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插入基因名称: | Mini-Tn, GFP cassette |
物种来源: | Pseudoalteromonas translucida |
基因突变: | NO |
克隆方法: | Gibson Cloning |
文献引用方法 |
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材料和方法部分: | pDonorGFP(Ptr) was a gift from Sheng Yang (Addgene plasmid # 175578 ; http://n2t.net/addgene:175578 ; RRID:Addgene_175578) |
参考文献部分: | Orthogonal CRISPR-associated transposases for parallel and multiplexed chromosomal integration. Yang S, Zhang Y, Xu J, Zhang J, Zhang J, Yang J, Jiang Y, Yang S. Nucleic Acids Res. 2021 Sep 3. pii: 6363765. doi: 10.1093/nar/gkab752. 10.1093/nar/gkab752 PubMed 34478496 |
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LOCUS pDonorGFP(Ptr) 4606 bp ds-DNA circular SYN 26-OCT-2021 DEFINITION DNA donor for transposition. ACCESSION . VERSION . KEYWORDS pDonorGFP(Ptr) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4606) AUTHORS Yang S, Zhang Y, Xu J, Zhang J, Zhang J, Yang J, Jiang Y, Yang S TITLE Orthogonal CRISPR-associated transposases for parallel and multiplexed chromosomal integration. JOURNAL Nucleic Acids Res. 2021 Sep 3. pii: 6363765. doi: 10.1093/nar/gkab752. PUBMED 34478496 REFERENCE 2 (bases 1 to 4606) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 26, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..4606 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(288..305) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(422..967) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(502..521) /label=pBR322ori-F /note="pBR322 origin, forward primer" RBS 1349..1371 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 1388..2095 /codon_start=1 /product="Aequoria victoria green fluorescent protein" /label=GFP /translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI CTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNY KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKANF KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFV TAAGITHGMDELYK" primer_bind complement(1412..1440) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 2035..2055 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 2189..2237 /label=fd terminator /note="central terminator from bacteriophage fd (Otsuka and Kunisawa, 1982)" terminator 2299..2347 /label=fd terminator /note="central terminator from bacteriophage fd (Otsuka and Kunisawa, 1982)" primer_bind complement(2416..2438) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3290..3309) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" CDS complement(3740..4399) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 4314..4333 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(4400..4502) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" ORIGIN 1 ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt tagtgtatga tggtgttttt 61 gaggtgctcc agtggcttct gtttctatca gctgtccctc ctgttcagct actgacgggg 121 tggtgcgtaa cggcaaaagc accgccggac atcagcgcta gcggagtgta tactggctta 181 ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct 241 gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac 301 tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat 361 ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt 421 ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt 481 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc 541 gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt 601 ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg 661 cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 721 aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga 781 gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg 841 cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa 901 accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg 961 atctcaagaa gatcatctta ttaatatacg tagtatctac gatacgtagt atctacgata 1021 cgtaggaggt tgagaagtca tttaataagg ccactgttaa acgtagatat atctacgcgt 1081 agatatatct acgatacgta gcggccgacg cgctgggcta cgtcttaatg ggcagcgccc 1141 acatacgcag cgatttccaa cagcttgagg cggaagcttt aatgcgggtc ctttgggtgt 1201 tgtttgaagt ataagttgac atatctgtac taaaagatgg cataaattgg aagtgtaagg 1261 tggcatagtc tagtatttaa ccaaatggtt aaatggttga ctcaccggga attccatggt 1321 cgacatgcat aagcttccta ggaataattt tgtttaactt taagaaggag atatacatat 1381 ggctagcaaa ggagaagaac tcttcactgg agttgtccca attcttgttg aattagatgg 1441 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg 1501 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact 1561 tgtcactact ttcacttatg gtgttcagtg cttttcaaga tacccggatc atatgaaacg 1621 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt 1681 caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt 1741 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttgggcacaa 1801 attggaatac aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg 1861 aatcaaagct aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga 1921 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta 1981 cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct 2041 tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat acaaaggatc 2101 cggctgctaa tcgagttaat taaactagtg agctcggtac ccggggatcc tctagaggtc 2161 gaaattcacc tcgaaagcaa gctgataaac cgatacaatt aaaggctcct tttggagcct 2221 ttttttttgg agattttcaa cgtgaaaaaa ttattattcg caattccaag ctaattcacc 2281 tcgaaagcaa gctgataaac cgatacaatt aaaggctcct tttggagcct ttttttttgg 2341 agattttcaa cgtgaaaaaa ttattattcg caattccaag ctctgcctcg cgcgtttcgg 2401 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 2461 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 2521 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatgggctcg atcccctcgg 2581 atccaattgg tcttaatttt ccttaattat ttttaaagtt agactgattt agacttggaa 2641 aagcttaatg attggagagc taaattgact aatagtattc agtcaagttt aattagtttt 2701 aagcgatcca tgcataacta tttctgtaca atgctatatt ttcaccaatt aataattttt 2761 aatcaagcct acattatgaa atatactata cccatttgaa ctcttctatt tgtaccattg 2821 tcggtagcaa aaacttatgg ggttttgaac gtcacttaaa ttgtaagcat ttgcgatgga 2881 ggcgcgttta gagtcaacct tgattctgat atgctccgaa tttttggtaa gaatataagt 2941 gtgagagtag ctaatgtgga tacgcctgag ttaagggaaa aatgtgaaaa tgaaataact 3001 cgttatcatg caaagtgact aaggttataa tcttccgttt atggcacata gcagccaact 3061 aaacttgaca gtatttttat gtggttggct ttataaaacc agcatttggt aacattatgc 3121 caatttttac ttcaatatta tgccaacata cactacacta acggagctgt agcacaataa 3181 gctcgtttgt acttatgcca acttatactt caaacaacat tggggtggat cctctacgcc 3241 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc 3301 gacatcaccg atggggaaga tcgggcctgc caccataccc acgccgaaac aagcgccctg 3361 caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac 3421 atctgtatta acgaagcgct aaccgttttt atcaggctct gggaggcaga ataaatgatc 3481 atatcgtcaa ttattacctc cacggggaga gcctgagcaa actggcctca ggcatttgag 3541 aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa 3601 gcggctattt aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc 3661 tgccattcat ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc 3721 aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt 3781 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca 3841 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga 3901 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg 3961 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt 4021 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac 4081 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac 4141 tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca 4201 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg 4261 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg 4321 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc 4381 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa 4441 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat 4501 caacgtctca ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag 4561 gatttattta ttctgcgaag tgatcttccg tcacaggtat ttattc //
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