质粒名称: | pSEVA-gRic6T |
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出品公司: | Addgene |
目录编号: | 106401 |
存储实验室: | Sheng Yang |
载体骨架基本信息 |
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载体名称: | pSEVA644 pTargetF |
完整质粒大小: | 4500 bp |
载体类型: | 细菌表达 |
载体抗性: | 庆大霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 未知 |
插入基因信息 |
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插入基因名称: | sgRNA |
物种来源: | Synthetic |
插入大小: | 96 bp |
启动子: | pj23119 |
克隆方法: | 未知 |
5’测序引物 | 未知 |
文献引用方法 |
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材料和方法部分: | pSEVA-gRic6T was a gift from Sheng Yang (Addgene plasmid # 106401 ; http://n2t.net/addgene:106401 ; RRID:Addgene_106401) |
参考文献部分: | Genome editing and transcriptional repression in Pseudomonas putida KT2440 via the type II CRISPR system. Sun J, Wang Q, Jiang Y, Wen Z, Yang L, Wu J, Yang S. Microb Cell Fact. 2018 Mar 13;17(1):41. doi: 10.1186/s12934-018-0887-x. 10.1186/s12934-018-0887-x PubMed 29534717 |
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pSEVA-gRic6T载体质粒基本信息
pSEVA-gRic6T质粒图谱载体图谱和pSEVA-gRic6T载体序列质粒序列多克隆位点信息
pSEVA-gRic6T质粒载体简介
pSEVA-gRic6T质粒序列载体序列
LOCUS pSEVA-gRic6T 4521 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION E.coli-Pseudomonas putida KT2440 shuttle plasmid containing sgRNA targeting the KT2440 genome, and homologous arms.. ACCESSION . VERSION . KEYWORDS pSEVA-gRic6T SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4521) AUTHORS Sun J, Wang Q, Jiang Y, Wen Z, Yang L, Wu J, Yang S TITLE Genome editing and transcriptional repression in Pseudomonas putida KT2440 via the type II CRISPR system. JOURNAL Microb Cell Fact. 2018 Mar 13;17(1):41. doi: 10.1186/s12934-018-0887-x. PUBMED 29534717 REFERENCE 2 (bases 1 to 4521) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..4521 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 25..59 /label=J23119 promoter /note="bacterial promoter (Registry of Standard Biological Parts BBa_J23119)" misc_RNA 80..155 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" regulatory 233..242 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" terminator 1247..1341 /label=lambda t0 terminator /note="transcription terminator from phage lambda" promoter 1372..1400 /gene="intI1 (promoter lies within the coding sequence)" /label=Pc promoter /note="class 1 integron promoter" oriT 2314..2422 /note="incP origin of transfer" rep_origin 2516..3104 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3005..3024 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3158..3991) /codon_start=1 /product="replication protein for the broad-host-range plasmid pRO1600 from Pseudomonas aeruginosa " /label=pRO1600 Rep /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA" rep_origin 4005..4356 /label=pRO1600 oriV /note="broad-host-range origin of replication from Pseudomonas aeruginosa plasmid pRO1600; requires the pRO1600 Rep protein for replication (West et al., 1994)" terminator 4429..4515 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 ttaattaatt gacaccatga attcttgaca gctagctcag tcctaggtat aatgctagca 61 ggcagaaaat cgcaatcgtg ttttagagct agaaatagca agttaaaata aggctagtcc 121 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttgaggagct cggtacccgg 181 ggatccggca gctctaccgc atacaccttg gccttcaaac gctgggcggc ctccatggcg 241 gcgttgaccg tgcgcgggtc ggagtagtgg ctcttgagca cggccacgct ctgcgtttcg 301 tccactcgag acagcttcag tacgtgctcg aacatctggg tcagttgtgc attgctcacc 361 ggcatcgggg ttctcctggg tggcgttgac aggggcgcac catggcagtg cctggtgctg 421 catctcggtg caaaataata gtgtgtacac cctacttgtg cagatgaaaa cgcatcctgc 481 cagcgagcgc aagtgatctg ttcaaacgtt acctatccac acattgacca gtatttacgg 541 taacaatcaa ccggatttcg tttcgatcag ctgtttgaaa aatattttca gctggctctt 601 ttcaaaggtc atttgtagcg tatacgctct aaatcgtcag gcggttgagc cgctggccac 661 ctatcagaac aagaggagac atacccgcac cttcgtcgct ggtggtaacg tcagcgccaa 721 tggcatccgc cagcattacc tgcgctatgg cggcaagggc catgcgctga tcctggtgcc 781 gggcatcacc agcccagcga tcacctgggg ctttgtcgcc gagcgccttg ggcattattt 841 cgacacctat gtgctggatg tgcgcgggcg tggcttgtct tcaagcggcc ccgacctgga 901 ctacggcacc gacgcttgtg ctgcggatat tcccgccttt gccgccgcgc tgggcctgga 961 tagctaccat ctgctcggtc actcgatggg ggcgcgtttt gccatccgcg cagccgccca 1021 aggggcaccg ggactgcagc ggctggtgct ggtcgaccca ccggtgtccg ggccgggccg 1081 ccgcgcctac ccgagcaagt tgccgtggta cgtcgattcg atccgccaag ccacggttgg 1141 catgagcggc gatgacatgc gcgccttctg tgccacctgg agcgacctgc aggcatgcaa 1201 gcttgcggcc gcgtcgtgac tgggaaaacc ctggcgacta gtcttggact cctgttgata 1261 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg 1321 gcgtttttta ttggtgagaa tccaggggtc cccaataatt acgatttaaa tttgacataa 1381 gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga 1441 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac 1501 tgtttttttg tacagcctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt 1561 cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca 1621 aagttaggtg gctcaagtat gggcatcatt cgcacatgta ggctcggccc tgaccaagtc 1681 aaatccatgc gggctgctct tgatcttttc ggtcgtgagt tcggagacgt agccacctac 1741 tcccaacatc agccggactc cgattacctc gggaacttgc tccgtagtaa gacattcatc 1801 gcgcttgctg ccttcgacca agaagcggtt gttggcgctc tcgcggctta cgttctgccc 1861 aagtttgagc agccgcgtag tgagatctat atctatgatc tcgcagtctc cggagagcac 1921 cggaggcagg gcattgccac cgcgctcatc aatctcctca agcatgaggc caacgcgctt 1981 ggtgcttatg tgatctacgt gcaagcagat tacggtgacg atcccgcagt ggctctctat 2041 acaaagttgg gcatacggga agaagtgatg cactttgata tcgacccaag taccgccacc 2101 taacaattcg ttcaagccga gatcgacttc ccggccgcgg agttgttcgg taaattggac 2161 aacggtccgc gcgttgtcct tttccgctgc ataaccctgc ttcggggtca ttatagcgat 2221 tttttcggta tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt 2281 agactttcct tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc 2341 cgcgagcggg tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa 2401 tcctgctctg cgaggctggc cgtaggccgg ccgataatct catgaccaaa atcccttaac 2461 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 2521 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 2581 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 2641 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga 2701 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 2761 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 2821 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 2881 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 2941 aggcggacag gcatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 3001 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 3061 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 3121 ccgtgaaagg caggccggtc cgtggtggcc acggcctcta ggccagatcc agcggcatct 3181 gggttagtcg agcgcgggcc gcttcccatg tctcaccagg gcgagcctgt ttcgcgatct 3241 cagcatctga aatcttcccg gccttgcgct tcgctggggc cttacccacc gccttggcgg 3301 gcttcttcgg tccaaaactg aacaacagat gtgtgacctt gcgcccggtc tttcgctgcg 3361 cccactccac ctgtagcggg ctgtgctcgt tgatctgcgt cacggctgga tcaagcactc 3421 gcaacttgaa gtccttgatc gagggatacc ggccttccag ttgaaaccac tttcgcagct 3481 ggtcaatttc tatttcgcgc tggccgatgc tgtcccattg catgagcagc tcgtaaagcc 3541 tgatcgcgtg ggtgctgtcc atcttggcca cgtcagccaa ggcgtatttg gtgaactgtt 3601 tggtgagttc cgtcaggtac ggcagcatgt ctttggtgaa cctgagttct acacggccct 3661 caccctcccg gtagatgatt gtttgcaccc agccggtaat catcacactc ggtcttttcc 3721 ccttgccatt gggctcttgg gttaaccgga cttcccgccg tttcaggcgc agggccgctt 3781 ctttgagctg gttgtaggaa gattcgatag ggacacccgc catcgtcgct atgtcctccg 3841 ccgtcactga atacatcact tcatcggtga caggctcgct cctcttcacc tggctaatac 3901 aggccagaac gatccgctgt tcctgaacac tgaggcgata cgcggcctcg accagggcat 3961 tgcttttgta aaccattggg ggtgaggcca cgttcgacat tccttgtgta taaggggaca 4021 ctgtatctgc gtcccacaat acaacaaatc cgtcccttta caacaacaaa tccgtccctt 4081 cttaacaaca aatccgtccc ttaatggcaa caaatccgtc cctttttaaa ctctacaggc 4141 cacggattac gtggcctgta gacgtcctaa aaggtttaaa agggaaaagg aagaaaaggg 4201 tggaaacgca aaaaacgcac cactacgtgg ccccgttggg gccgcatttg tgcccctgaa 4261 ggggcggggg aggcgtctgg gcaatccccg ttttaccagt cccctatcgc cgcctgagag 4321 ggcgcaggaa gcgagtaatc agggtatcga ggcggattca cccttggcgt ccaaccagcg 4381 gcaccagcgg cgcctgagag gggcgcgccc agctgtctag ggcggcggat ttgtcctact 4441 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc 4501 ctttcgtttt atttgatgcc t //
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